Vous êtes sur la page 1sur 70

GENETICA HUMANA.

Human genetic diseases and normal variations can be placed into one of five categories: 1. single gene disorders (diseases or traits where the phenotypes are largely determined by the action, or lack of action, of mutations at individual loci); 2. multifactorial traits (diseases or variations where the phenotypes are strongly influenced by the action of mutant alleles at several loci acting in concert); 3. chromosomal abnormalities (diseases where the phenotypes are largely determined by physical changes in chromosomal structure - deletion, inversion, translocation, insertion, rings, etc., in chromosome number trisomy or monosomy, or in chromosome origin - uniparental disomy); 4. mitochondrial inheritance (diseases where the phenotypes are affected by mutations of mitochondrial DNA); and 5. diseases of unknown etiology that seem to "run in families." About 1% of the approximately 4 million annual live births in the United States will have a single gene disorder that will be serious enough to require special medical treatment or hospital care. Each of these single gene disorders, often called Mendelian traits or diseases, is relatively uncommon. The frequency often varies with ethnic background, with each ethnic group having one or more Mendelian traits in high frequency when compared to the other ethnic groups. For example, cystic fibrosis has a frequency of about 1/2000 births in Americans descended from western European Caucasians but is much rarer in Americans of western African descent while sickle cell anemia has a frequency of about 1/600 births in Americans of western African descent but is much rarer in Caucasians. Greeks and Italians of Mediterranean descent have a high frequency of thalassemia; Eastern European Jews have a high frequency of Tay-Sachs disease; French Canadians from Quebec have a high frequency of tyrosinemia, all when compared to other ethnic groups. It has been estimated that each of us, each "normal" member of the human race is carrying between 1 and 8 mutations which, if found in the homozygous state would result in the expression of a Mendelian disease. Since we each have between 50,000 and 100,000 genes (loci) it is unlikely that any two unrelated individuals would be carrying the same mutations, even if they are from the same ethnic background, thus most of our offspring are not suffering from a genetic disease. Most Mendelian diseases are rare, affecting about 1/10,000 to

1/100,000 live births as an order of magnitude estimate. In total they will add to the 1% of live births mentioned above. Mendelian traits, or single gene disorders, fall into 5 categories or modes of inheritance based on where the gene for the trait is located and how many copies of the mutant allele are required to express the phenotype: 1. autosomal recessive inheritance (the locus is on an autosomal chromosome and both alleles must be mutant alleles to express the phenotype) 2. autosomal dominant inheritance (the locus is on an autosomal chromosome and only one mutant allele is required for expression of the phenotype) 3. X-linked recessive inheritance (the locus is on the X chromosome and both alleles must be mutant alleles to express the phenotype in females) 4. X-linked dominant inheritance (the locus is on the X chromosome and only one mutant allele is required for expression of the phenotype in females), and 5. mitochondrial inheritance (the locus is on the mitochondrial "chromosome"). Mendel based his laws on mathematical probabilities that allowed predictions of resulting phenotypes when certain crosses were made in the garden pea. When he published in 1866, the discovery of the chromosomal basis of inheritance (meiosis and gametogenesis) by Sutton, Boveri, and others was still a generation away. Therefore, there was no physical basis for explaining the Mendelian segregation ratios. The discoveries of Sutton, Boveri, and others allowed a reexamination of Mendel's apparently forgotten publication. In 1900, Correns, DeVries, and Tschermak, all independently "rediscovered" Mendel's laws of segregation, and by 1902 the first human Mendelian "inborn error of metabolism", alcaptonuria, was found by Sir Archibald Garrod. Mendel's laws are grounded in the chromosomal movements in meiosis, gametogenesis, and fertilization. Understanding the fundamental processes of cell division is the key to understanding Mendelian genetics.

MITOSIS and MEIOSIS


Mitosis is the process of cell division that is responsible for the development of the individual from the zygote (fertilized egg) to maturity

(approximately 10 cells). It is the process by which the somatic cells divide and maintain the same chromosomal complement. Each chromosome duplicates forming two chromatids connected to a single centromere, the centromeres line up on the metaphase plate without the homologous pairing and recombination found in meiosis (except for sister chromatid exchange of identical DNA information in mitosis), and the centromere divides as each chromatid now becomes a daughter chromosome at anaphase of cell division. Mitosis is the process by which two identical daughter cells with identical DNA complements are formed from one progenitor cell. Mutations can arise during DNA replication in mitosis, just as they do in meiosis. These mutations, and their consequences in somatic cell diseases, such as cancer, are discussed in the molecular genetics lecture portion of this course. Most mitotic divisions, and consequently the fastest rate of growth, occurs before birth in the relatively protected environment of the uterus. Most of us only increase 15 to 30 times our birth weight (2 or 2 times) from birth to maturity, but from conception to birth our weight increases many fold. Consequently, most genetic diseases are expressed at birth or during early development, although some late onset human diseases, and somatic cell diseases, do occur.
14 4 5

Gelehrter, Collins, and Ginsburg, Chapter 2, should be read for a complete description of the events and importance of mitosis .

MEIOSIS and GAMETOGENESIS


Each somatic cell of a normal individual contains two copies of each of the 22 autosomal chromosomes, one of paternal origin and one of maternal origin, and either an X from the mother and an X from the father if the individual is female or an X from the mother and a Y from the father if the individual is male. This is called the diploid (2 copy) state. During gametogenesis, the formation of the gametes (ova in females and sperm in males), this diploid state is reduced to the haploid (1 copy) state through the process of cell division called meiosis. Meiosis consists of two consecutive cytoplasmic divisions with only one DNA replication. In some texts meiosis will be explained as two divisions, a reduction division followed by a mitotic division but this is a misnomer. Meiosis is one continuous process from beginning to end.

This diagram shows a general summary of two pairs of chromosomes going through meiosis. Only the nucleus and the centrioles are shown. In A, the chromosomal DNA is already replicated and the homologous chromosomes are partially paired. In B, pairing is completed but the two chromatids of each chromosome have not yet condensed enough to be visible. In C, both chromotids of each chromosome are visible and recombination (chiasma), or crossing-over, between chromatids of the homologous chromosomes are evident. In D, the chiasmata (pl. of chiasma) are being resolved and the homologous centromeres are lining up on the metaphase plate. E represents anaphase of the first meiotic division. The centromeres of homologous chromosomes are moving to the poles without dividing, thus separating the maternal centromere from the paternal centromere along with their associated chromosomes that have recombined. In F, the centromeres each of the haploid chromosomes with its two chromatids are migrating to the metaphase plate. G shows the centromeres dividing and moving toward the poles in early anaphase of the second meiotic division. H demonstrates the nuclei of the 4 haploid products that result from the meiotic division of one initial diploid cell. In humans, none of the four haploid products is identical, since recombination occurs at least once for each chromatid, but they all contain the same amount of DNA and each contains 23 chromosomes. The chromosomal movements in oogenesis and spermatogenesis in humans will be covered more completely in the section on chromosomal abnormalities. It is presented here to show the chromosomal movements required to fulfill Mendel's laws. Gelehrter, Collins, and Ginsburg, Chapter 2 should be read for a more complete introduction to meiosis and the structure of human chromosomes. Mendel assumed that the traits he was studying were determined by what he called unit characters. We call these unit characters alleles. Alleles are the alternative forms of a gene, often called the locus or specific site on the chromosome where the gene resides. Mendel's law of segregation states that

during gametogenesis these alternative forms, alleles, segregate into different gametes and are never found in the same gamete. The chromosomal movements in meiosis assure this. The above sketch reviews the chromosomal movements of first meiotic division. [A], represents two homologous chromosomes in a cell that is going to enter meiosis, one chromosome was inherited from the mother and one inherited from the father. Each chromosome contains a single double stranded DNA molecule. Each has a different allele at a particular locus. [B], the chromosomes have duplicated, forming two chromatids (two double-stranded DNA molecules) and paired at the metaphase plate in the first division of meiosis. [C], the homologous chromosomes have separated at the first division. Notice that the alleles are destined to go into separate gametes. The effects of recombination are not shown. Mendel's law of independent assortment states that unit characters for different traits, traits controlled by genes of different chromosomes assort independently. That is, if a gene on chromosome 1 has two alleles, a and b, and a gene on chromosome 2 has two alleles, c and d, the combinations a and c, a and d, b and c, and b and d, are all equally likely. There is no preference for a to be with either c or d. Since chromosmes 1 and 2 line up on the metaphase plate independently at the first meiotic division, with equal chance of the maternal or paternal homolog going to one pole for each chromosome, these combinations have an equal chance of occurring. Thus, alleles of genes that lie on different chromosomes assort independently of one another. These two laws, the law of segregation and the law of independent assortment, are the basis of Mendelian inheritance. [Go on to next lesson] or [Return to top of this page] or [Return to the Course Outline] uizzes on Mendelian Inheritance are available on-line at our secure Mallard site. Click here and the UIC WWW Identification Service will ask for your netid and then your password (these are the same as those you use for email.) Once the Mallard page loads you can access the quizzes by clicking on the Lessons Page link (also the third icon from the top of the navigation bar)

or the Current Lesson link (also the fourth icon from the top of the

navigation bar).

he study of inherited Mendelian traits in humans must rely on observations made while working with individual families. Classical cross fertilization breeding experiments as performed by Mendel are not allowed in humans! Human geneticists are not allowed to selectively breed for the traits they wish to study! One of most powerful tools in human genetic studies is pedigree analysis. When human geneticists first began to publish family studies, they used a variety of symbols and conventions. Now there are agreed upon standards for the construction of pedigrees.

Males are always represented by square symbols, females with circular symbols. A line drawn between a square and a circle represents a mating of that male and female. Two lines drawn between a square and a circle indicate a consanguineous mating, the two individuals are related, usually second cousins or closer relatives. When possible, the square should be placed on the left and the circle on the right of the mating line. Generations are connected by a vertical line extending down from the mating line to the next generation. Children of a mating are connected to a horizontal line, called the sibship line, by short vertical lines. The children of a sibship are always listed in order of birth, the oldest being on the left. Sometimes to simplify a pedigree only one parent is shown, the other is omitted. This neither signifies parthenogenic development nor does it signify divinely inspired conception, it merely means the parent left out is not from the family being studied and is genotypically homozygous normal for the trait being studied. Normal individuals are

represented by an open square or circle, depending upon the gender, and affected individuals by a solid square or circle. Each generation is numbered to the left of the sibship line with Roman Numerals. Individuals in each generation are numbered sequentially, beginning on the left, with Arabic Numerals. For example the third individual in the second generation would be identified as individual II-3. For more information on the construction and interpretation of pedigrees consult Gelehrter, Collins, and Ginsburg, 2nd edition, Chapter 3.

AUTOSOMAL DOMINANT INHERITANCE


The pattern of autosomal dominant inheritance is perhaps the easiest type of Mendelian inheritance to recognize in a pedigree. One dose of the mutant gene, one mutant allele, is all that is required for the expression of the phenotype. There are three reasons why an individual with an autosomal dominant disease should always be considered as being a heterozygote until proven otherwise: 1. The disease is usually rare, with only about 1/10,000 individuals affected as an order of magnitude. To produce a homozygote, two affected heterozygotes would have to mate. This probability is 1/1,000,000 and then they would have only a 1/4 chance of having a homozygous affected offspring. Affected individuals are most likely to come from affected by normal matings. The normal parent is homozygous recessive, thus assuring that each product of the mating has at least one normal gene. 2. In the extremely rare instances where two affected individuals have mated, the homozygous affected individuals usually are so severely affected they are not compatible with life. The exceptions are the autosomal dominant diseases caused by the somatic expansion of trinucleotide repeat sequences (e.g., Huntington's disease) that we will study later. 3. The mating of very closely related individuals, the most likely way for two affected individuals to know each other, is forbidden in our society. With the understanding that almost all affected individuals are heterozygotes, and that in most matings involving a person with an autosomal dominant trait the other partner will be homozygous normal, there are four hallmarks of autosomal dominant inheritance.

1. Except for new mutations, which are rare in nature and extremely rare

on examination pedigrees, and the complexities of incomplete penetrance to be discussed later, every affected individual has an affected biological parent. There is no skipping of generations. 2. Males and females have an equally likely chance of inheriting the mutant allele and being affected. The recurrence risk of each child of an affected parent is 1/2. 3. Normal siblings of affected individuals do not transmit the trait to their offspring.
4. The defective product of the gene is usually a structural protein,

not an enzyme. Structural proteins are usually defective when one of the allelic products is nonfunctional; enzymes usually require both allelic products to be nonfunctional to produce a mutant phenotype.

THE PUNNET SQUARE


In 1910, Punnett developed a simple method of depicting the possible genotypes one could get from various matings. We call it the Punnett Square. Its use in predicting the genotypic ratios in the offspring is illustrated below: Suppose a father is heterozygous for an autosomal dominant gene, with allele D, the mutant dominant allele, and allele d, the recessive normal allele. He can produce two types of gametes, D and d. Suppose also his wife is homozygous normal, having both d alleles. The Punnett Square is constructed as follows:

One gamete comes from each parent to produce the genotype of the offspring. Two out of the four possible combinations are affected; two out of four are normal.

AUTOSOMAL DOMINANT INHERITANCE

Sample P e di g r e e

The family represented by Pedigree 1 is a good example of how autosomal dominant diseases appear in a pedigree. Each of the four hallmarks of autosomal dominant inheritance are fulfilled. Each affected individual has an affected parent; there is no skipping of generations. Males and females are equally likely to be affected. About 1/2 of the offspring of an affected individual are affected (the recurrence risk is 1/2). Normal siblings (II-3) of affected individuals have all normal offspring. Low density lipoprotein receptors are structural proteins or polypeptides, not enzymes. If III-1, an affected female, were to produce a child that child would have a 1/2 chance of being normal and a 1/2 chance of being affected. If her normal brother, III-2, were to produce a child that child would have a nearly 0 chance of being affected.

uizzes on Mendelian Inheritance are available on-line at our secure Mallard site. Click here and the UIC WWW Identification Service will ask for your netid and then your password (these are the same as those you use for email.) Once the Mallard page loads you can access the quizzes by clicking on the Lessons Page link (also the third icon from the top of the navigation bar) or the Current Lesson link (also the fourth icon from the top of the navigation bar).

MENDELIAN INHERITANCE

AUTOSOMAL DOMINANT INHERITANCE

ince every mutant allele for an autosomal dominant disease is expressed, and by definition a disease is a deleterious phenotype, how do autosomal dominant diseases stay in the population? Shouldn't they be eliminated by natural selection against deleterious phenotypes? There appears to be four phenomena that maintain these deleterious alleles in the population: 1. 2. 3. 4. Variable Expressivity Late Onset High Recurrent Mutation Rate Incomplete Penetrance

VARIABLE EXPRESSIVITY
One example of variable expressivity is Marfan syndrome. Marfan syndrome is an autosomal dominant disease caused by a mutation in collagen formation. It affects about 1/60,000 live births. Symptoms of Marfan syndrome include skeletal, optical, and cardiovascular abnormalities. Skeletal abnormalities include arachnodactyly (long fingers and toes), extreme lengthening of the long bones, scoliosis, rib and sternum abnormalities, among others. Optical abnormalities almost always include ectopia lentis, a dislocation of the lens into the anterior chamber of the eye. Cardiovascular abnormalities may be numerous and include possible dissecting aneurysms, which are largely responsible for the shorter life span of Marfan syndrome patients as a group. Each patient may express all of the symptoms, or only a few. That is variable expressivity. Each patient with the mutant allele for Marfan syndrome expresses at least one of the symptoms, but the physician may have to look closely. Almost all are taller than average, but a lot of nonMarfan individuals are tall. Almost all have long fingers, but so do a lot of non-Marfan persons. Some may be very mildly affected and lead normal lives while others, more severely affected, have a shorter life expectancy. The disease is maintained in the population through recurrent mutations and the matings of less severely affected individuals with normal individuals. The extent of severity of affected does not affect the severity of expression in the next generation, that is, the offspring of mildly affected individuals range from mildly affected to severely affected, with equal probability. For a more complete description and other examples of variable expressivity see Gelehrter, Collins, and Ginsburg, 2nd ed., Pages 29 and 30.

LATE ONSET

Some autosomal dominant diseases do not express themselves until later in life, well beyond the reproductive years. The individuals who will develop the disease have passed the mutant allele along to their offspring before they themselves know they are affected. In some cases even grandchildren are born before the affected grandparent shows the first signs of the disease. Huntington disease, sometimes called Huntington's Chorea because of the choreic movements expressed as the disease progresses, is a good example of a late onset disease. Age of onset varies from the teens to the late sixties, with a mean age of onset between ages 35 and 45. Nearly 100% of the individuals born with the defective allele will develop the disease by the time they are 70. The disease is progressive with death usually occurring between four and twenty-five years after the first symptoms develop. Emotional changes often are the first symptoms. At the gene level, it is caused by the expansion of an unstable trinucleotide repeat sequence, CAG, in the coding region of the gene. What is inherited at birth in Huntington disease is a gene with several repeats and the instability that allows somatic recombination and extension. Somatic mutations introduced by the expansion of trinucleotide repeat sequences do not have to occur in coding regions to produce a mutant allele. Other diseases, such as myotonic dystrophy, an autosomal dominant disease where expression is delayed, result from molecular defects at the gene level that are caused by the expansion of unstable trinucleotide sequences. (In the case of myotonic dystrophy the sequence is CTG.) However, this unstable sequence lies in a non-translated region of the gene. In both diseases the size of the inherited expansion correlates to the age of onset or the severity of disease, but is not absolutely predictable on an individual basis. One cannot sequence the gene and precisely predict the age of onset of Huntington disease.

HIGH RECURRENT MUTATION RATE


Achondroplasia is one of the major causes of dwarfism. Motor skills may not develop as quickly as their normal siblings, but intelligence is not reduced. It occurs in about 1/10,000 live births. Like many autosomal dominant diseases, individuals homozygous for the mutant allele do not survive to term. Almost 85% of the cases are the result of new mutations, where both parents have a normal phenotype. The mutation rate for achondroplasia may be as much as 10 times the "normal" mutation rate in humans. This high recurrent mutation is largely responsible for keeping the mutant gene in the population at its present rate. Several other autosomal dominant genetic diseases have high recurrent mutation rates but achondroplasia is probably the best known.

INCOMPLETE PENETRANCE
Incomplete penetrance should never be confused with variable expressivity. In diseases with variable expressivity the patient always expresses some of the symptoms of the disease and varies from very mildly affected to very severely affected. In autosomal dominant diseases with incomplete penetrance, the person either expresses the disease phenotype or he/she doesn't. Incomplete penetrance and variable expressivity are phenomena associated only with dominant inheritance, never with recessive inheritance. The following pedigree illustrates incomplete penetrance in a known autosomal dominant disease.

In the above pedigree, there is ample evidence for autosomal dominant inheritance:

The disease is passed from the father (II-3) to the son (III-5), this never happens with X-linked traits. The disease occurs in three consecutive generations, this never happens with recessive traits. Males and females are affected, with roughly the same probability.

However, II-1 does not express the disease. He must have inherited the mutant allele because he passed it on to two children, III-1 and III-3. II1 is a classical example of incomplete penetrance, he has the allele for the disease but he does not express it.

Late onset, high recurrent mutation rate, variable expressivity, and incomplete penetrance only occur in autosomal dominant diseases, never in recessive diseases unless there is medical intervention to prevent the development of symptoms in neonatal children.

AUTOSOMAL RECESSIVE INHERITANCE


he first, and most important, thing to remember about autosomal recessive inheritance is that most, if not all, affected individuals have parents with normal phenotypes. Why? Suppose the disease affects one in ten thousand live births, a good order of magnitude estimate for most autosomal recessive diseases. That would make the heterozygote frequency in the population one in fifty (see population genetics for calculations). The likelihood of two affected persons mating would be 1/10,000 x 1/10,000 or 1/100,000,000. By chance alone there might be two such matings in the Unites States, but no more than 2. The likelihood of an affected and a heterozygote mating would be 1/10,000 x 1/50 x 2(since either parent could be the affected) or 1/250,000. The likelihood of two heterozygotes (heterozygotes are usually called "carriers") mating is 1/50 x 1/50 or 1/2500, more than 99% of all possible matings. The Punnett Square for autosomal recessive diseases with an affected child in the family almost always looks like the following:

Where the father and mother are both Dd (dd is the recessive affected individual, Dd the heterozygous carrier individual, and DD the homozygous normal individual). The Punnet Square shows the origin of the famous Mendelian ration of 3/4 normal to 1/4 affected. For most autosomal recessive diseases, but not all, the heterozygote cannot be distinguished from the normal homozygote. In the normal phenotype categories of offspring in the above Punnett Square (Dd and DDproduce the same normal phenotype),

please note that two of the three are heterozygotes (carriers); one of the three is homozygous normal. Within the normal siblings of an affected individual the probability of being a carrier is 2/3. There are five hallmarks of autosomal recessive inheritance: 1. Males and females are equally likely to be affected. 2. On average, the recurrence risk to the unborn sibling of an affected individual is 1/4. 3. The trait is characteristically found in siblings, not parents of affected or the offspring of affected. 4. Parents of affected children may be related. The rarer the trait in the general population, the more likely a consanguineous mating is involved.

5. 5. The trait may appear as an isolated (sporadic) event in small sibships.

The above pedigree illustrates four of the five hallmarks of autosomal recessive inheritance. I-1 and I-2 are unrelated, yet they produced an affected offspring (affected offspring have normal parents). By chance, they both must have been carriers. Even though II-2 is affected, she produced no affected offspring (trait appears in siblings, not parents or offspring). By far the most probable genotype for an individual from outside the family (II-1)

is homozygous normal. III-1, III-2 and III-3 are all obligate carriers (heterozygotes), since they are not affected but could only have inherited the recessive gene from II-2 II-3, II-5, and II-6 each have a 2/3 chance of being a carrier and a 1/3 chance of being homozygous normal. They are not affected, but they come from a carrier x carrier mating. II-4 and II-7 have a high probability of being homozygous normal since they are from outside the family. III-4, III-5, III-6, III-7, III-8, and III-9 all have a 1/3 chance of being carriers and a 2/3 chance of being homozygous normal. One parent of each is probably homozygous normal, the other has a 2/3 chance of being a carrier and a 1 in 2 chance of passing on the recessive allele if they were a carrier. When consanguinity is involved, i.e., matings between related individuals, in the production of an affected child the assignment of probabilities changes, especially in the rarer autosomal recessive diseases. Since these relatively rare autosomal recessive diseases would have disease frequencies of 1/10,000 live births or less, the carrier frequency in the general population would not exceed 1/50. Normal individuals from the general population would have a probability of at least 49 to 1 of being homozygous. Consanguinity introduces the possibility of one founding parent being a carrier, with the recessive allele being passed through carrier offspring and meeting itself to produce an affected homozygous offspring some generations later. When an affected child is produced as the result of a consanguineous mating, those individuals in the direct line of descent are most probably carriers and those from outside the family are most probably normal homozygotes. In the following pedigree, V-1 is affected with an autosomal recessive disease. Her parents are second cousins, a legal marriage in most states. IV-1 and IV-2 must both be carriers since they produced an affected child. (The child must have received a recessive allele from each of her parents.) III-2 is an obligate carrier. Her father was affected, and hence, a homozygote for the recessive allele. III-5 must also be heterozygotes since IV2 had to get her recessive allele from one of her parent, and the chance of III-6 being a carrier is less than 1 in 50. I-1 and I-2 must both have been carriers since they produced an affected offspring, II-1.

Normally one never considers the possibility of two unrelated individuals both being carriers unless there is evidence to the contrary. Here I-1 and I-2 are the exception to the rule. There is evidence that both must be carriers. Before he had any children, II-5 had a 2/3 chance of being a carrier and a 1/3 chance of being homozygous normal (The normal siblings of affected rule explained above). But III-5 had to get her recessive allele from someone, and her other parent, II-6 had at most a 1/50 chance before her children were born. One can compare the two probabilities and calculate that in at least 100 out of 103 times II-5 will be the carrier. This is so close to 1 that for practical purposes one can say he is the carrier. In rare autosomal recessive diseases, when consanguinity is involved, those individuals in the direct line of descent within the family are considered to be carriers and those individuals from outside the family are considered homozygous normal unless there is evidence to the contrary. What can we say about the carrier probabilities of the individuals within the family that are not in the direct line of descent to the affected child? In the above pedigree, III-1 must be a carrier. She is not affected, but she must have received a recessive allele from her father (II-1) who is homozygous recessive. II-3 and II-4 each have a 2/3 chance of being a carrier since they are phenotypically normal and come from a heterozygote x heterozygote mating. III-6 has a one in two chance of being a carrier. His father is a carrier (see above calculations) and his mother is from outside the family. In human genetics counseling it doesn't help your patient to understand and make decisions when you are quoting statistics to the second or third decimal point. 49/100 or 0.488 are close to one half. One out of two, or one half, is a number your patient is more likely to understand and remember. You may get into trouble for using certainty (zero or 100%), or even ridiculous

numbers like 1/1,000,000, when something doesn't go as you predicted, but not for rounding off to the nearest fraction. Always make your counseling calculations as accurately as possible, but make sure they are presented in a form that is something your patient will comprehend.

MENDELIAN INHERITANCE

X-LINKED INHERITANCE
hen the locus for a gene for a particular trait or disease lies on the X chromosome, the disease is said to be X-linked. The inheritance pattern for X-linked inheritance differs from autosomal inheritance only because the X chromosome has no homologous chromosome in the male, the male has an X and a Y chromosome. Very few genes have been discovered on the Y chromosome. The inheritance pattern follows the pattern of segregation of the X and Y chromosomes in meiosis and fertilization. A male child always gets his X from one of his mother's two X's and his Y chromosome from his father. Xlinked genes are never passed from father to son. A female child always gets the father's X chromosome and one of the two X's of the mother. An affected female must have an affected father. Males are always hemizygous for X linked traits, that is, they can never be heterozygoses or homozygotes. They are never carriers. A single dose of a mutant allele will produce a mutant phenotype in the male, whether the mutation is dominant or recessive. On the other hand, females must be either homozygous for the normal allele, heterozygous, or homozygous for the mutant allele, just as they are for autosomal loci.

X-LINKED DOMINANT INHERITANCE


When an X-linked gene is said to express dominant inheritance, it means that a single dose of the mutant allele will affect the phenotype of the female. A recessive X-linked gene requires two doses of the mutant allele to affect the female phenotype. The following are the hallmarks of X-linked dominant inheritance:

The trait is never passed from father to son. All daughters of an affected male and a normal female are affected. All sons of an affected male and a normal female are normal.

Matings of affected females and normal males produce 1/2 the sons affected and 1/2 the daughters affected. Males are usually more severely affected than females. The trait may be lethal in males. In the general population, females are more likely to be affected than males, even if the disease is not lethal in males.

The following Punnett Squares explain the first three hallmarks of Xlinked dominant inheritance. X represents the X chromosome with the normal allele, XA represents the X chromosome with the mutant dominant allele, and Y represents the Y chromosome. Note that the affected father never passes the trait to his sons but passes it to all of his daughters, since the heterozygote is affected for dominant traits. On the other hand, an affected female passes the disease to half of her daughters and half of her sons.
Father's Gametes XA Y A XX XY XXA XY Father's Gametes Mother's Gametes X X
A

Mother's Gametes

X X

X XAX XX

Y XAY XY

Figure 1. Affected father x normal mother.

Figure 2. Affected mother x normal father.

Males are usually more severely affected than females because in each affected female there is one normal allele producing a normal gene product and one mutant allele producing the non-functioning product, while in each affected male there is only the mutant allele with its non-functioning product and the Y chromosome, no normal gene product at all. Affected females are more prevalent in the general population because the female has two X chromosomes, either of which could carry the mutant allele, while the male only has one X chromosome as a target for the mutant allele. When the disease is no more deleterious in males than it is in females, females are about twice as likely to be affected as males. As shown in Pedigree 5 below, Xlinked dominant inheritance has a unique heritability pattern.

The key for determining if a dominant trait is X-linked or autosomal is to look at the offspring of the mating of an affected male and a normal female. If the affected male has an affected son, then the disease is not X-linked. All of his daughters must also be affected if the disease is X-linked. In Pedigree 5,

both of these conditions are met. What happens when males are so severely affected that they can't reproduce? Suppose they are so severely affected they never survive to term, then what happens? This is not uncommon in X-linked dominant diseases. There are no affected males to test for X-linked dominant inheritance to see if the produce all affected daughters and no affected sons. Pedigree 6 shows the effects of such a disease in a family. There are no affected males, only affected females, in the population. Living females outnumber living males two to one when the mother is affected. The ratio in the offspring of affected females is: 1 affected female: 1 normal female: 1 normal male.

Pedigree 6.
You will note that in Pedigree 6 there have also been several spontaneous abortions in the offspring of affected females. Normally, in the general population of us normal couples, one in six recognized pregnancies results in a spontaneous abortion. Here the ratio is much higher. Presumably many of the spontaneous abortions shown in Pedigree 6 are males that would have been affected had they survived to term.

X-LINKED RECESSIVE INHERITANCE


veryone has heard of some X-linked recessive disease even though they are, in general, rare. Hemophilia, Duchenne muscular dystrophy, Becker muscular dystrophy, and Lesch-Nyhan syndrome are relatively rare in most populations, but because of advances in molecular genetics they receive attention in the media. More common traits, such as glucose-6phosphate dehydrogenase deficience or color blindness, may occur frequently enough in some populations to produce a few affected females. However, their effect on individuals is rarely life threatening and medical intervention is not needed. Pedigree 7 shows one typical inheritance pattern for a rare Xlinked recessive disease.

In Pedigree 7 above, which of the following females is least likely to be a heterozygote for the rare X-linked recessive gene, III1, III-3, or III-5? The answer of course is III-3. III-1 and III-5 each have a 1/2 chance of being a carrier but III-3 has almost a 0 chance of being a carrier. Why? Let's look at the PunnettSquares for X-linked recessive inheritance.
Affected Father's Genotype XA Y Normal Father's Genotype X Y

Normal Mother's Genotype

X X

XXA XXA

XY XY

All daughters carriers, all sons normal.

Carrier XA XAY XAY Mother's X XX XY Genotype Half of sons affected, half of daughters carriers.

In Pedigree 7, II-2 and II-5 are both carriers, their father was affected and passed on his only X chromosome to his daughters. II-3 cannot be a carrier for two reasons. First, males are either affected or normal, never carriers. Second, he didn't inherit his father's X chromosome. He inherited his father's Y chromosome. III-3 couldn't have been a carrier since neither her father nor her mother had the mutant gene. What are the hallmarks of X-linked recessive inheritance?

As with any X-linked trait, the disease is never passed from father to son. Males are much more likely to be affected than females. If affected males cannot reproduce, only males will be affected. All affected males in a family are related through their mothers. Trait or disease is typically passed from an affected grandfather, through his carrier daughters, to half of his grandsons.

Counseling in X-linked recessive diseases is a bit more complex than it is in autosomal recessive diseases. In X-linked recessive diseases, Bayes theorem, or Bayesian probability must be used to accurately calculate carrier probabilities. In some pedigrees these probabilities change as new information appears. Sometimes we use Bayesian probability without recognizing it. Consider the following: A patient of yours is getting married and comes to you for counseling. She has a brother with a rare X-linked recessive disease. Her mother's father also had the disease. She wants to know the probability of her being a carrier of the disease and the probability that she will pass the disease to her children. What is your advice? Being a reasonably good human geneticist, you tell her that her mother was a carrier and that she has a one chance in two of being a carrier, depending upon which of her mother's X chromosomes she inherited. You also explain that if she is a carrier she will pass the affected X to her son one

half of the time, but that her daughters will not be affected because they will always get a normal X from their father. Excellent advice! But now suppose the conditions change. Her first child is a boy and he is affected. Now what is the probability that she is a carrier? Of course this probability changes from 1/2 to 1. That is the basis for Bayesian probability. The individual undergoing counseling has an a priori calculated probability of being a carrier (not certainty) and later developments alter this probability. Now suppose this patient who came to you for counseling had a normal son, not an affected son. Shouldn't her probability of being a carrier be changed? Yes it should, but it is more complex than moving from 1/2 to 1 as it did with the affected son. The following is an example of how probabilities change with normal sons.
Probability of NonCarrier Carrier 1/2 1/2

At Birth

At birth she had an equal chance of being a carrier or a non-carrier, her mother was a known carrier. What is the probability of 1 normal son, given that she is a carrier? Answer: 1/2 What is the probability of 1 normal son, given that she is a non-carrier? Answer: 1
Probability of NonCarrier Carrier At Birth 1/2 1/2 One Normal Son 1/2 1

The joint probability of both events happening is the product of each separate event. The probability of being a carrier and also having one normal son is 1/2 x 1/2. The probability of being a non-carrier and having a normal son is 1/2 x 1.
Probability of NonCarrier Carrier At Birth 1/2 1/2 One Normal Son 1/2 1 Joint Probability 1/4 1/2

Since your patient is either a carrier or a non-carrier, we need to calculate the relative probability of each possibility. To do this we must make the joint probabilities add to 1. This is done by dividing each joint probability by the sum of the joint probabilities.
Probability of Carrier Non-Carrier At Birth 1/2 1/2 One Normal Son 1/2 1 Joint Probability 1/4 1/2 Relative Probability (1/4)/(1/4+1/2)=1/3 (1/2)/(1/4+1/2)=2/3

The relative probability of your patient being a carrier after the birth of one normal son is 1/3 and the relative probability of her not being a carrier is now 2/3. Just as the probability changed after the birth of an affected son, it also changes after the birth of a normal son. Now, if she has another son and he is normal, her probability of being a carrier and producing two normal sons would be 1/2 x 1/2 x 1/2. The joint probability is now 1/8 of being a carrier. The joint probability of not being a carrier would be 1/2 x 1 x 1. It would remain at 1/2. The relative probability of being a carrier would drop to 1/5. The relative probability of not being a carrier would increase to 4/5. However, if she were to have a third son and he were affected, all of this Bayesian probability calculation would not be necessary. She would be a known carrier. If she were a non-carrier, her affected son would have to be a new mutation. This is not an impossibility for X linked recessive diseases because it only requires one mutational event to be expressed in males, but when it is factored in the Bayesian probability tables above as a probability of 10-5, it shifts the probability of being a carrier to almost certainty.

Pedigree 8 shows a family with a rare X-linked recessive trait. For which of the following females should Bayesian probability be used to calculate the carrier probability, II-2, II-5, III-2, III-5, or III-7? II-2 and II-5 are both certain carriers, their father was affected and they produced affected sons. III-2 and III-5 each had a 1/2 chance of being carriers at birth. One of them has one normal son, the other has had two normal sons. These are Bayesian probabilities. III-7 is related to the affected individuals of the family through her father. She has an almost 0 chance of being a carrier.

Pedigree 9 demonstrates the one other common use of Bayesian probability in counseling. Two requirements must be met. The disease must be a genetic lethal, that is, individuals with the disease cannot reproduce. An example of such a disease is Duchenne muscular dystrophy. The proband must be the only affected individual in the family. When these two conditions are met, as they are in Pedigree 9, there is a reasonable possibility that the affected individual, III-1, is a new mutation. What is the probability that any female in the population where there is no history of the disease is a carrier of the Duchenne muscular dystrophy gene? She could be a new mutation. If the mutation rate is u, her probability of being a new mutation would be 2u because she has two X chromosomes and each could be mutated. Her mother could have been a new mutation (2u) with a 1/2 chance of passing it to her daughter. Her grandmother could have been the new mutation (2u), with a 1/4 chance of passing the mutant allele to her granddaughter. (Note that the father has no part in the equation since he is normal - affected males cannot reproduce.) Her great-grandmother could have been the new mutation (2u) with a 1/8 chance of passing the mutation to her great-granddaughter, etc. Since these are all independent ways to produce a carrier female, the probabilities sum: 2u + 1/2(2u) + 1/4(2u) + 1/8(2u) + 1/16(2u) + ... = 4u A female with no history of the disease in her family has a 4u chance of being a carrier before her first son is born. She has a 1-4u chance of not being a carrier. This is close enough to 1 for the purposes of our Bayesian calculations. The Bayesian table for II-2 in Pedigree 9 looks like this:
Carrier 4u 1/2 2u 2/3 Probability of Non-Carrier 1-4u=1 u (III-1 was a new mutation) u 1/3

At Birth Probability of one affected child (III-1) Joint Probability Relative Probability

Even though II-3 has produced an affected son, under the above two conditions she still has only a 2/3 chance of being a carrier. 1/3 of all Duchenne muscular dystrophy children born into families with no history of the disease must be new mutations. This is true only of X-linked recessive diseases where the affected males cannot reproduce.

MENDELIAN INHERITANCE

SEX LIMITED INHERITANCE


n some X-linked recessive diseases, such as Duchenne muscular dystrophy, expression of the disease phenotype is limited exclusively to males. In some X-linked dominant traits, such as incontinentia pigmenti or orofaciodigital syndrome (OFD 1), expression is limited to females, males do not survive to term. However, the expression of a disease in only one gender does not necessarily imply that the disease is X-linked. There are autosomal diseases that are limited to expression in only one sex. Precocious puberty and beard growth are factors expressed only in males. The hereditary form of prolapsed uterus is expressed only in females. These are called sex limited traits.

MITOCHONDRIAL INHERITANCE
The DNA of mitochondria contains about ten genes involved in oxidative phosphorylation, as well as a few other genes. This DNA is capable of mutation, so it is not surprising that a few human diseases have been found to be associated with mitochondrial inheritance. Leber optic atrophy is a classic example of a disease of mitochondrial DNA. The ovum, originating in the female, has about 100,000 copies of mitochondrial DNA; the sperm, originating in the male, has fewer than 100 copies, and these are probably lost at fertilization. Virtually all of ones mitochondria come from his, or her, mother. Affected fathers produce no affected offspring, while the offspring of affected mothers are all affected. Figure 3 below shows the typical mitochondrial inheritance pattern.

Figure 3 Mitochondrial inheritance pattern.

IMPRINTING

Prader-Willi syndrome affects between 1/10,000 and 1/30,000 live births. The study of this disease led to the discovery that, for some genes, the origin of the gene may be important. For some loci the gene inherited from the father acts differently from the gene inherited from the mother, even though they may have the same DNA. This phenomenon is called imprinting. About 75% of patients with Prader-Willi syndrome have a small deletion of the long arm of chromosome 15, a small piece of one chromosome 15 is missing while the homologous chromosome remains intact. When this deletion is on the paternal chromosome (the father's genes are missing) Prader-Willi syndrome results. When this deletion is on the maternal chromosome (the mother's genes are missing) Angelman syndrome results. The two diseases have very different clinical symptoms. The other 25% of Prader-Willi syndrome patients are almost all the result of uniparental disomy, a rare chromosomal event in which both chromosomes come from a single parent. (This will be covered later under chromosomal diseases.) When both chromosomes 15 are derived from the mother, Prader-Willi syndrome results. When both chromosomes 15 are derived from the father, Angelman syndrome results. For normal development an individual must inherit one copy of this chromosomal region from his or her father and one from his or her mother. Several other regions have been found to show uniparental disomy without this effect on the phenotype. Small deletions usually affect the phenotype but they produce the same phenotype whether of maternal or paternal origin. Through some unknown mechanism, the gene, or genes, involved in Prader-Willi and Angelman syndrome know their origin and behave according to that origin. At the present time we do not know whether this is a general phenomenon or not. It might be limited to this small region of chromosome 15. It might be quite wide spread. Imprinting represents an exception to Mendel's laws and remains an important area of research.

CHROMOSOMAL INHERITANCE

IMPORTANCE
here are about five million conceptions in the United States each year, give or take a few hundred thousand. Consider the fate of 10,000 randomly chosen from these five million. About 800 are chromosomally abnormal. Of these 800:

At least 140 are 45, X. They lack an X or a Y chromosome. At least 110 have an extra chromosome 16. At least 20 have an extra chromosome 18.

At least 40 have an extra chromosome 21. The rest have various different chromosomal abnormalities.

Of the 800 chromosomally abnormal conceptions, about 750 will abort spontaneously:

139 of the 140 who lack an X or a Y chromosome will abort spontaneously. All of those with an extra chromosome 16 will abort spontaneously. 19 of the 20 with an extra chromosome 18 will abort spontaneously. The survivor will have a very short life expectancy. 35 of the more than 40 with an extra chromosome 21 will abort spontaneously. The survivors will have Down syndrome. There are a variety of conceptions with other chromosomes extra. All will be aborted spontaneously.

The remaining 50 individuals with chromosomal abnormalities will make it to birth. Among them should be about:

1 with an extra chromosome 18 1 with a missing X or Y chromosome 10 with an extra chromosome 21 15 with an extra X or Y chromosome, and about 20 with abnormal chromosomal rearrangements of various sorts, at least 4 of which will result in Down syndrome.

Chromosomal abnormalities are an important component of medical practice. You will see examples of them in your work and your everyday life.

KARYOTYPE
Specific chromosomal abnormalities were very difficult to identify prior to 1956. In that year, Tjio and Levan published their method for visualizing the chromosomes and revolutionized cytogenetics. They made it possible to accurately count the chromosomes and determine in which of 7 groups of chromosomes the error occurred (Groups A through G). Their methods started nearly two decades where research on chromosomal abnormalities was the focus of human genetics. Shortly after their discovery, this intense study led to other advances in techniques that allowed for the identification of individual chromosomes by differential staining, resulting in unique banding patterns for each chromosome. Now even very small regions can be visualized. To identify chromosomes, they are arrested in late prophase or early metaphase of mitosis, when the chromosomes are duplicated and condensed, but the centromere has not yet divided. Each chromosome consists of two chromatids at this stage. They are stained, photographed and arranged in a particular

order, from

largest to smallest, in what is called a karyotype. An example of the karyotype of a normal male is shown in Figure 4 below. The chromosomes are grouped by size and location of the centromere (metacentric, submetacentric, and acrocentric).

Figure 4. Karyotype.
There is also a standard nomenclature for describing various karyotypic abnormalities. Refer to Gelehrter, Collins, and Ginsburg, 2nd ed., Chapter 8, for ideograms and chromosomal nomenclature. You will be responsible for all of the material in this chapter. Each chromosome consists of one double-stranded DNA molecule running the length of the chromosome, along with histones and other proteins. The DNA is arranged in chromatin loops that have general gene expression coordination. Two other distinct structures are essential, the centromere and the telomere. The centromere is the site of attachment of the spindle fiber. Without it the chromosome would not move in mitosis and meiosis, would be lost from the nucleus, and would be degraded by cytoplasmic enzymes. The telomeres are distinct structures at each end of the chromosome. They maintain the structural integrity of the chromosomal DNA. When a telomere is missing there is a strong tendency of the chromosomes to join with one another, or with pieces of one another, causing abnormal gene expression and aberrant chromosome structures.

CHROMOSOME REPLICATION
Chromosome replication is different in humans than it is in bacteria, although the method of DNA replication is the same. In bacteria there is one origin of replication, but the polymerase is very fast. In humans the polymerase copies only about 100 to 150 base pairs per second, but there are 20,000 to 100,000 origins of replication along the 46 chromosomes. Most of these origins are functional in embryogenesis when divisions are occurring very rapidly, but many fewer function in the adult. DNA synthesis begins at several origins of replication along a chromosome and moves in each direction until it meets the replicated strands from the next origin.

Figure 5.

Chromosome replication.
The chromosomes replicate in a predetermined sequence in mitosis, with each member of the homologous pair duplicating at the same time. The exception in somatic cells, not germ line cells, is that one of the X chromosomes of the female is always last of all the chromosomes to replicate. X replication in female somatic cells does not occur simultaneously in each of the homologs.

AUTOSOMAL ABNORMALITIES
Several autosomal abnormalities produce such serious changes in the phenotype that they are not compatible with life. Any ploidy (extra copies of all chromosomes, i.e., triploidy, tertaploidy, etc.) higher than diploid results in early death in utero. Trisomy, an extra copy of a single chromosome, except for chromosomes 13, 18, or 21, is not compatible with life. Trisomy 13 and Trisomy 18 each lead to early death, usually in the neonatal period of development if notin utero.

Meiotic nondisjunction, the failure of the chromosomes to disjoin and pass to opposite poles, in either the first or second meiotic division is the major cause of chromosomal abnormalities. The greatest percentage of these (75%) occurs in oogenesis, where the probability of nondisjunction increases with maternal age. Almost 80% occur in the first meiotic division. To understand a possible mechanism for these observations we need to review oogenesis and spermatogenesis.

Figure 6. Events in spermatogenesis. (Note the timing.)


Spermatogenesis begins at puberty and continues on a regular pace through the lifetime of the male, although spermatogonia do not divide as rapidly during later life. Males in their eighth decade have been documented fathers. The process from spermatogonium to mature sperm takes about 64 days, with each division evenly spaced at about 16 days, as shown in Figure 6. Spermatogenesis, with its many more cell divisions is more prone to single gene mutations than is oogenesis. There may be as much as a four or five fold increase in the mutation rate for some Mendelian traits in the sperm of older men. Oogenesis, on the other hand, is not associated with a higher mutation rate for Mendelian diseases as maternal age increases. Increasing maternal age is associated with a higher incidence of chromosomal abnormalities attributed to nondisjunction. The reason for this association is evident from Figure 7 below.

Figure 7. Events in oogenesis.


All of the oocytes that are ever going to develop in the female are present at birth and all have begun the first meiotic division before they are arrested in the dictyotene state for from 12 to 40 years. Then with each menstrual cycle, one oocyte begins to continue on through the first and second meiotic divisions. These divisions occur very rapidly. With an old mitotic apparatus it is very possible that mistakes in chromosomal movement and cell division will occur. In some ways spermatogenesis, with its liability for mutation, and oogenesis, with its liability for chromosomal aberrations, may be thought of as complementary, one protects against mutation while the other protects against chromosomal abnormalities.

MEIOTIC NONDISJUNCTION
Nondisjunction, the failure of the chromosomes to disjoin and move to opposite poles may affect as many as 25% of all ova and 2% of all sperm. Half of these abnormal gametes are nullisomy, half are disomy. Two copies of a chromosome pass into the same gamete, leaving the other gamete as nullisomy. At fertilization the zygote formed from the gamete with nullisomy gets one copy of the chromosome from the gamete of the other parent and becomes monosomic for that chromosome. Monosomy for an autosome is not compatible with life. At fertilization, the zygote formed from the gamete with

the two copies of the chromosome gets a third copy from the gamete of the other parent and become trisomic. As discussed earlier, most do not survive to term. Down syndrome, or trisomy 21, is the classic example of a human disease caused by autosomal nondisjunction where some, but not all, affected individuals do survive. It occurs in about 1/800 live births, which means around 5000 affected individuals are born each year in the United States. The disease occurs in all races and nationalities. Approximately 95% of these 5000 Down syndrome children are the result of meiotic nondisjunction. The error occurred in first meiotic division in about 80% of these individuals; the remaining 20% occurred in second meiotic division. Errors in oogenesis account for 75% of the births while about 25% are of paternal origin. For a more complete discussion of Down syndrome, including the effects of increasing maternal age, see Gelehrter, Collins, and Ginsburg, 2nd ed., Chapter 8. There are three other causes of Down syndrome besides simple trisomy 21. Mitotic nondisjunction, isochrome formation, and Robertsonian translocation can also produce Down syndrome.

MITOTIC NONDISJUNCTION (Mosaic)


A chimera is an individual that results from the fusing of two cell lines, from two zygotes, during development. Twins can, under extremely rare circumstances, be chimeras. A mosaic individual is one who originated as a single cell line and through some mitotic event develops two different cell lines. Mitotic nondisjunction is one of the events that will produce a mosaic individual. When mitotic nondisjunction of chromosome 21 occurs early in development of a female, two new cell lines develop, 45, XX, -21 and 47, XX, +21, in addition to the 46,XX founding cells. The monosomy 21 cell line does not survive. The karyotype of the mosaic female is written 46, XX/47,XX + 21. Of course there is an equal probability for the mosaic to arise in a male. The severity of affected of mosaic individuals depends upon how early the nondisjunction occurred and what cell lines developed from those early embryonic cells.

ROBERTSONIAN TRANSLOCATION
About 9% of Down syndrome children born to mothers who are less than 30 years of age will be the result of Robertsonian translocation. Down syndrome mothers under the age of 30 have a relatively low recurrence risk for a second trisomy 21 if the first affected child resulted from either meiotic or mitotic nondisjunction. However, the recurrence risk is much higher if the

affected child was the result of a Robertsonian translocation. Robertsonian translocations are limited to the acrocentric chromosomes, chromosomes 13, 14, 15, 21, and 22. These chromosomes all have short arms, the p arms, that largely are made up of the genes for ribosomal RNA. These short arms are usually called satellites. There are many copies of these genes and a person can function quite well if several are lost. These satellites have a great deal of homology from chromosome to chromosome, and they tend to associate during interphase and mitotic division. Occasionally they will exchange parts, and the long arm, the q arm of one chromosome will become attached to the q arm of another, with the loss of the two parms, or satellites. All that is lost is a centromere and some ribosomal RNA genes. (Small chromosomal structures get lost in mitosis because they do not attach to a spindle fiber.) Two q arms are now attached (translocated) to the same centromere. This is called a Robertsonian translocation. The formation of a Robertsonian translocation is shown in Figure 8 below.

Figure

8. Robertsonian translocation.
There is no effect on the individual in which this mitotic event occurs. He or she has a normal phenotype in all respects. Mitosis is unaffected. However, if the translocation occurs in a cell that is destined to be in the germ line, then trouble may arise when that cell tries to undergo meiosis. Pairing of homologous chromosomes at metaphase of the first meiotic division may create problems as described in Figure 9 below.

Figure 9. Pairing of homologous chromosomes.


Figure 9 shows that pairing at the first meiotic division can occur three ways, with equal chances for all. When it occurs as shown in A, one secondary oocyte or spermatocyte will get the 14q21q translocation chromosome and the other secondary oocyte or spermatocyte will get a normal 14 and a normal 21. When fertilization occurs, with a normal 14 and a normal 21 from the other parent, the zygotes formed from the gametes with pairing as in A will be either 45, XX or XY, -14, -21, +t(14q21q) a phenotypically normal balanced Robertsonian translocation carrier, or be a 46, XX or XY normal individual. When pairing at first meiotic division is as shown in B, one secondary oocyte or spermatocyte will get chromosome 14 only (no chromosome 21) and the other secondary spermatocyte or oocyte will get chromosome 14q21q and chromosome 21. At fertilization, again the zygote will get a normal 21 and a normal 14 from the other parent. When the gamete from B that got only chromosome 14 unites with a normal gamete, there is monosomy for chromosome 21. That results in very early embryonic death and spontaneous abortion. When the gamete from B that got the 14q21q chromosome plus chromosome 21 unites with a normal gamete, one gets the prober dose of necessary genetic information for chromosome 14 (14 from the normal gamete and 14q of the translocation chromosome) but one has three copies of the genetic information of chromosome 21(21 from the normal gamete, and a 21 and a 21q from the translocation). This results in Down syndrome. When, by chance the chromosomes line up as at C, both gametic products are lethal early in development. One will get only chromosome 21 and will be lacking a chromosome 14. Monosomy for chromosome 14 is lethal. One will get a 14 and a 14q21q, resulting in a zygote that will be trisomic for chromosome 14. This trisomy is also lethal. From a carrier of a Robertsonian translocation there are 6 types of gametes, only three of which will produce a fetus capable of surviving to

term. Of these three combinations who may survive, one should be normal, one should be phenotypically normal but be a carrier of the translocation, and one should have Down syndrome. In actual practice, if the mother is the carrier of the translocation only about 11% of the children have Down syndrome, the remaining 89% non-Down syndrome children are equally divided between normals and balanced translocation carriers. Trisomy 21 embryos do not survive as well as those with the normal amount of genetic information. If the father is a carrier of the translocation, only about 2% of the offspring will have Down syndrome, the remaining 98% being equally divided between normal and balanced translocation carriers. Because Robertsonian translocation is responsible for about 9% of the Down syndrome children born to mothers under the age of 30, it is important to karyotype the child to determine if the child is the result of a Robertsonian translocation or simple meiotic nondisjunction. As one can see, the counseling of the parents is entirely different. Robertsonian translocations are passed from generation to generation, and with this type of inheritance Down syndrome may "run in families."

ISOCHROME FORMATION
Isochrome, or 21q21q, may result from a Robertsonian translocation between the two 21 chromosomes during mitosis in the germ line, or it may result from an improper mitotic division of the centromere, where the centromere divides transversely rather than longitudinally. Either of these is a rare event, but they do happen. When that happens it produces a karyotype, 45, XX or XY (depending upon the sex of the individual) -21,-21,+I(21q). Both q arms of chromosome 21 are attached to the same centromere. In gametogenesis, the gametes from an isochrome 21 individual get either the isochrome (21q21q) or they are missing chromosome 21 entirely. The zygotes are then either monosomy 21, which is lethal, or they have a normal 21 and a 21q21q chromosome, resulting in Down syndrome. All of the viable offspring of an isochrome 21 individual will have Down syndrome.

CHROMOSOMAL INHERITANCE

SEX CHROMOSOME ABNORMALITIES


n humans the normal female has two X chromosomes, while the normal male has only one X chromosome. If a gene is on the X chromosome, isn't it logical that there would be twice the gene product in females than there is in males? This was a question that remained unanswered for many years. From biochemical measurements there seemed to be the roughly the same amount of gene product in both males and females. The phenomenon was called "dosage compensation." Somehow the gene

knew to compensate for the sex of the individual, either make half as much product if it found itself in a female or make twice as much product if it was in a male. Dosage compensation was explained by the discoveries of Mary Lyon. LYON HYPOTHESIS The Lyon hypothesis states that during early development, about the 100 cell stage in humans, one of the X chromosomes in a female gets turned off and this is maintained in all descendant cells of the clone. Which of the two X chromosomes gets turned off in each of the 100 cells is purely a random event except where one of the X chromosomes is abnormal (deletion, insertion, inversion, etc.) An abnormal X is always turned off. However, if there is a translocation between an X chromosome and an autosome, the normal X is turned off and the translocation X remains active. In other words, under normal conditions a female is a mosaic in each tissue derived from somatic cells. If she is a heterozygote for a gene that is on the X chromosome and controls an enzyme, say G6PD, on the average half of her cells will express one allelic product of the G6PD gene and the other half of her cells will express the other allelic product. No somatic cell will express both alleles. This explains the phenomenon of dosage compensation. Only one gene product is produced in each cell of the female and only one gene product is expressed in each cell of the male. However, for some reason both X chromosomes remain active in female germ line cells. BARR BODIES The inactive X usually lies along the edge of the interphase nucleus in a highly condensed state. It is always the last to replicate. In 1948, before the discoveries of Lyon, Barr and Bertram found that in the interphase nucleus of female cat neurons there were a significant number of cells that had one "darkly staining body" lying along the edge of the nucleus, but they never found a "darkly staining body" in the neurons of male cats. Similar "darkly staining bodies" are found in buccal epithelial cells of human females, although they can usually be found in only 30% to 40% of the cells. Normal males never express these "Barr bodies." In all cases, the number of Barr bodies is one less than the number of X chromosomes in an individual. One Barr body means the individual has two X chromosomes, two Barr bodies means the individual has three X chromosomes, etc. We now know that the "darkly staining" Barr body is the condensed, inactive X chromosome. We also know that not all regions of the X chromosome in a female are turned off. There is a blood group locus, Xg, which is near the end of the X chromosome. Heterozygotes for this blood group express both allelic products

on each erythrocyte. So there are exceptions to the Lyon hypothesis, but it holds true as a general phenomenon. Since X inactivation is a random event, sometimes more than half of the cells will have, by chance, the same X inactivated. Rarely, but not as rarely as mutation, so many cells of the heterozygote will share the same inactive X that the female phenotype will appear to be that of the homozygote. TURNER SYNDROME Just as simple meiotic nondisjunction is the leading cause of autosomal chromosome abnormalities, so is nondisjunction the leading cause of the X and Y abnormalities. In autosomal abnormalities an increase in nondisjunction was associated with increasing maternal age. In sex chromosome abnormalities, one additional source of nondisjunction can be identified, the problem of X and Y pairing at first meiotic division in spermatogenesis. The X and Y have homology only in a small region (called the pseudo autosomal region) which lies near one end of each chromosome. Rather than pair along their entire length, pairing (and possible recombination) occur only in this small region. At first meiotic division in the male, pairing of X and Y looks more like end to end pairing than longitudinal pairing. This undoubtably adds to the frequency of nondisjunction. Turner syndrome (45,X) is the most frequent chromosomal abnormality. It is found in more than 7% of all spontaneous abortions. As it affects only about 1/2500 live female births, only about 2% of the recognized 45,X embryos survive to term, 98% are lost. The nondisjunction that results in a 45,X female can occur at either meiotic division in either spermatogenesis or oogenesis, but about 80% are the result of paternal nondisjunction. Turner syndrome individuals can also result from early mitotic nondisjunction, being mosaic 46,XX/45,X. The phenotype of Turner syndrome individuals differs significantly from the normal female, even though the normal female has only one functioning X chromosome in each cell. The events of embryogenesis during the time both X chromosomes are functioning in the female must be critical, as well as those few regions of the X chromosome that are not inactivated. Refer to Gelehrter, Collins, and Ginsburg, 2nd ed., Chapter 8, for complete descriptions of the phenotypes, and for further details on the sex chromosome abnormalities, including fragile X. KLINEFELTER SYNDROME Klinefelter syndrome (47, XXY) occurs in about 1/850 male births. In the human, the presence of one Y chromosome produces male secondary sex

characteristics in the absence of specific mutations for sex determining loci. Since the child must get his Y chromosome from his father, the nondisjunction that produces a Klinefelter syndrome child could occur in either meiotic division of the mother, but could only occur in the first meiotic division in the father. If nondisjunction occurred in the father, the zygote would have to get both an X and a Y chromosome in the same sperm. In spermatogenesis this could only result from a mistake in first division. Second meiotic division of spermatogenesis separates either the two X chromatids into different gametes or the two Y chromatids into different gametes. XYY & XXX SYNDROMES XYY syndrome occurs in about 1/1000 male births. It can only result from nondisjunction in the second meiotic division of spermatogenesis. Even though it affects only 1/1000 men, it is found in almost 1/50 males in prison populations. Aggressive behavior and less intelligence than siblings are often included in phenotypic descriptions of these 47,XYY individuals. XXX syndrome (47,XXX) has such a normal phenotype that it is not usually classified as a disease or even recognized unless there are reproductive problems with spontaneous abortions. However, as a general rule, as the number of X chromosomes increases past the diploid state, mental deficiencies increase.

NON-MEIOTIC CHROMOSOMAL ABNORMALITIES


> number of gross chromosomal changes can arise that results either in altered gene control or in difficulties during meiosis or mitosis. Inversions, non-Robertsonian translocations, and ring chromosomes all produce, or have the potential for producing, altered phenotypes. Each of them is rare when compared to nondisjunction. INVERSIONS Inversions involve two chromosomal breaks and rejoining, with the broken piece reincorporated in the opposite orientation from which it naturally occurs. When they include the centromere, they are called pericentric inversions. When they do not include the centromere, they are called paracentric inversions. Both types of inversions arise in mitotic cells. If they arise in precursors of the gametes, they may produce abnormal genomes as they progress through meiosis. Recombination between homologous chromosomes is a necessary part of every normal meiosis. The probability for nondisjunction is greatly increased if there is no recombination. However,

recombination between a chromosome with an inversion and its normal homolog may result in two abnormal chromosomes being produced. The results of a paracentric inversion going through meiosis, with a recombination within the inversion, are shown in Figure 10. Of the four gametic products, one is normal, one has the inversion, one has an acentric chromosome, and one has a dicentric chromosome. The acentric chromosome cannot survive. The dicentric chromosome may be pulled apart during mitosis, with a random loss or gain of genetic material.

Figure 10. Paracentric Inversion.


The results of a pericentric inversion going through meiosis, with a recombination within the inversion is shown in figure 11. Again, four different gametic products are produced, one normal, one has the inversion, and two have duplicated portions and deleted portions. The effect on the phenotype is almost always deleterious, but the magnitude of the effect depends upon the size of the duplications and deletions, and where they occur.

Figure 11. Pericentric Inversion.


RING CHROMOSOMES Ring chromosomes result from the loss of the telomeres, with a rejoining of the ends of the same chromosome. When a telomere is lost there is a strong tendency for the chromosome to unite with a similar fragment lacking a telomere. Ring chromosomes cause no problem until there is a sister chromatid exchange. Sister chromatid exchange is a rather common mitotic event. It can be detected in five or six chromosomes at each mitotic division Since the two chromatids are identical in all respects, there is no gain or loss of genetic material. However, when a sister chromatid exchange occurs in a ring chromosome, a double chromatid is produce that is dicentric. Dicentric rings have the same problem going through mitosis as the dicentric chromosomes of paracentric inversions. When the two centromeres line up on opposite sides of the metaphase plate, they migrate to opposite poles, and the chromosome is randomly broken, with loss or gain of genetic material. This always produces a deleterious effect on the phenotype. NON-ROBERTSONIAN TRANSLOCATIONS Non-Robertsonian translocations, translocations between chromosomes other than those of the D or G groups, always have trouble going through first

meiotic division. One half of the gametes of a person with a translocation will involve duplications of some genetic material and deletions of other portions of the chromosome. These are always deleterious, usually resulting in severely affected individuals. UNIPARENTAL DISOMY Included in the discussion of Mendelian traits was a section on imprinting in Prader-Willi syndrome and Angelman syndrome. 75% of the time these diseases are caused by deletion within chromosome 15, more precisely 15q1113. If the deletion occurs in the chromosome that came from the father (only maternal genes present), Prader-Willi syndrome results. If the deletion occurs in the chromosome that came from the mother(only paternal genes present), Angelman syndrome results. The two syndromes are not at the same genetic locus, but are controlled by genes that are within this small region of chromosome 15. They have very different phenotypes.
Prader-Willi Neonatal hypotonia & developmental deficiency Severe obesity Short stature Hypogonadism Mild to moderate mental retardation with learning disabilities Small hands and feet Angelman Ataxia, seizures Hyperactivity Severe mental retardation Absence of speech Inappropriate laughter (Happy puppet syndrome)

In about 25% of Prader-Willi syndrome children, and about 2% of Angelman syndrome children, the cause of the disease is not deletion but uniparental disomy, both copies of chromosome 15 came from one parent. In the case of Prader-Willi syndrome both chromosomes came from the mother. In the case of Angelman syndrome, both chromosomes came from the father. In about 1/30,000 conceptuses the zygote probably was a trisomy 15, a lethal genetic condition, and early in mitotic development one chromosome 15 was lost, restoring the embryo to the diploid state. If the nondisjunction that produced the trisomy 15 occurred in the mother, with the subsequent loss of the paternal chromosome 15, Prader-Willi syndrome results. If the trisomy resulted from paternal nondisjunction with subsequent loss of maternal chromosome 15, Angelman syndrome results. These two mechanisms, deletion and uniparental disomy, account for virtually all of the Prader-Willi syndrome patients. Angelman syndrome can also result from mutations within the gene (or genes) that produce nonfunctional gene products.

Although it is a rare event, uniparental disomy has been documented for other autosomal chromosomes. Except for the unexpected expression of an autosomal recessive disease (heterozygote x homozygous normal mating), no detectable phenotypic effect has been found.

MULTIFACTORIAL INHERITANCE

IMPORTANCE
ultifactorial inheritance is responsible for the greatest number of individuals that will need special care or hospitalization because of genetic diseases. Up to 10% of newborn children will express a multifactorial disease at some time in their life. Atopic reactions, diabetes, cancer, spina bifida/anencephaly, pyloric stenosis, cleft lip, cleft palate, congenital hip dysplasia, club foot, and a host of other diseases all result from multifactorial inheritance. Some of these diseases occur more frequently in males. Others occur more frequently in females. Environmental factors as well as genetic factors are involved.

REGRESSION TO THE MEAN


Multifactorial inheritance was first studied by Galton, a close relative of Darwin and a contemporary of Mendel. Galton established the principle of what he termed "regression to mediocrity." Mendel studied discontinuous characters, green peas vs. yellow peas, tall vs. dwarf, etc. There was no overlap of phenotype in Mendel's studies. Characters fit into one of two classes. There was no blending in the heterozygote. On the other hand, Galton studied the inheritance of continuous characters, height in humans, intelligence in humans, etc. Galton noticed that extremely tall fathers tended to have sons shorter than themselves, and extremely short fathers tended to have sons taller than themselves. "Tallness" or "shortness" didn't breed true like they did in Mendel's pea experiments. The offspring seemed to regress to the median, or "mediocrity." Figure 12 shows the correlation between the father's height and the height of the son.

Figure 12. A representation of Galtons studies on the inheritance of height. If the sons height were determined only by the fathers height, the correlation should be that of the solid line. The dashed line is what is observed. Galton called this "regression to mediocrity."
If the son's height were completely determined by the father's height, the correlation would be as shown by the solid blue line. What is observed is shown by the dashed red line. The height of the father and the average height of the son are related, but the average height of the son always regresses toward the mean. That is understandable if there is no dominance. The son only gets half of his father's genes; the other half comes from his mother. When comparing height differences between men and women, women are, on average, 3 inches shorter. A woman with a certain number of "tall" genes will be, on average, 3 inches shorter than a man with the same number. When that difference is taken into account, there is no selective bias in matings for tallness in human populations. It is true than men tend to marry women who are shorter than themselves, but that is a phenotypic difference, not a genotypic difference. Since the wives of taller than average men tend to represent the general population of women, they will not have, on the average, as many "tall" genes to pass on to their offspring as their husbands. Hence, the son will receive half of the father's "tall" genes, on average, and half of the

mother's "tall" genes, on average, but his total genes for "tallness," on average, will be less than his father's. Shorter than average males have fewer "tall" genes than average, but they are still as tall as an average female, even though the average female has more "tall" genes. Their sons, on average, will be taller than their fathers because their mothers have, on average, more "tall" genes to give to their sons than their husbands have. On average, the son will have more "tall" genes than his father. What holds true for height also holds true for other quantitative traits, such as intelligence. This is what worried Galton. He was a very intelligent member of British aristocracy who was interested in genetics as a way to maintain intelligence in his family. He was really the founder of the eugenics movement. His findings must have been very discouraging for him.

POLYGENIC INHERITANCE
For many years the argument raged between the "Mendelians" and the "Galtonians" as to which of the two paradigms was the correct one for human inheritance. There was no question that Mendelian inheritance was correct for some diseases, but these were rare, affecting only a small portion of the population. They were considered trivial by the Galtonians. On the other hand, the inheritance of quantitative traits could not be used to predict outcomes, only average estimates measured in large population studies. Mendelians considered the study of quantitative traits to be trivial because they had no predictive value. R. A. Fisher resolved the dispute by showing that the inheritance of quantitative traits can be reduced to Mendelian inheritance at many loci. Fisher's argument went as follows: Consider the following: One locus for height, with three alleles. Allele h2 adds 2 inches to the average 68-inch height. Allele h0 neither adds nor subtracts from the average height of 68 inches. And allele h- subtracts 2 inches from the average height. Suppose h0 is twice as frequent as either h2or h-. The Punnett square for the population would be as follows:
FATHER'S GAMETES h2 2h0 hh2,h2 2(h2,h0) h2,h72" 70" 68" 2(h2,h0) 4(h0,h0) 2(h-,h0) 70" 68" 66" h2,h2(h-,h0) h-,h68" 66" 64"

h2 MOTHER'S 2h0 GAMETES h-

When this is expressed in tabular form, it looks like the histogram in Figure 13.

Figure 13. The distribution of height in a population if it were determined by one locus with three alleles as described in the text.
If a second locus, called the tall locus, or t, is also involved in height, with three alleles as above, one adding two inches, one neither adding nor subtracting from the phenotype, and one subtracting 2 inches, with the neutral allele occurring twice as frequently as the either of the others, the histogram becomes that of Figure 14.

Figure 14. The distribution of height in a population if were determined by two loci, each with three alleles as described in the text.

THE MULTIFACTORIAL MODEL


As more loci are included, this binomial distribution quickly approaches the Gaussian distribution, or the bell-shaped normal curve, observed with human quantitative traits. Three loci, each with three alleles, are enough to produce population frequencies indistinguishable from a normal curve. The multifactorial model is then: 1. Several, but not an unlimited number, loci are involved in the expression of the trait. 2. There is no dominance or recessivity at each of these loci. 3. The loci act in concert in an additive fashion, each adding or detracting a small amount from the phenotype. 4. The environment interacts with the genotype to produce the final phenotype. As an example of 4. above, women are, on average, three inches shorter than men with the same genome. Environmental factors (hormones) affect the final phenotype. Not all human traits that show a continuous distribution in the population are multifactorial traits. Any bimodal distribution is not controlled by

multifactorial expression. It is more likely to be under the control of a single dominant/recessive gene with modifying environmental factors. Multifactorial traits all show a unimodal bell-shaped distribution.

CONCORDANCE
Twin studies, although limited by complicating factors, provide the best source for separating genetic contributions to the trait being studied from environmental influences. Monozygous (identical) twins have the same genome, but not the exact environmental factors, especially if they were raised apart. The concordance rate in monozygotic twins can be compared to the concordance rate in dizygotic (fraternal) twins to estimate the genetic component (heritability) of the trait. If the trait is truly 100% genetic, as it is for total fingerprint ridge count in humans, monozygotic twins will be 100% concordant while dizygotic twins, having, on average, only half their genes in common, will have a lower concordance rate. If the trait under study is 100% environmental, monozygotic twins and dizygotic twins will have the same concordance rate. The concordance rate for a disease is calculated as follows: Concordance Rate = [Both Affected / (One Affected + Both Affected)] x 100 For quantitative traits, means and variances have to be substituted and the calculations are beyond the scope of this introductory course.

THRESHOLD MODEL OF DISEASE


If multifactorial traits are quantitative traits with continuous distribution, how can they control diseases, such as cleft lip or spina bifida? One either has the disease or doesn't. There is no intermediate. I'm glad you asked that question. Multifactorial diseases are best explained by the threshold model shown in Figure 15

Figure 15. The threshold model for multifactorial traits. Below the threshold the trait is not expressed. Individuals above the threshold have the disease.
As the number of multifactorial genes for the trait increases, the liability for the disease increases. When it reaches a threshold, the liability is so great that abnormality, what we call disease, results. For example, consider the development of the cleft palate. Early in embryonic development the palatal arches are in a vertical position. Through embryonic and fetal development the head grows larger, making the arches farther apart, the tongue increases in size, making it more difficult to move, and the arches themselves are growing and turning horizontal. There is a critical stage in development by which the two arches must meet and fuse. Head growth, tongue growth, and palatal arch growth are all subject to many genetic and environmental factors. If the two arches start to grow in time, grow at the proper rate, and begin to move soon enough to the horizontal they will meet and fuse in spite of head size and tongue growth. The result is no cleft palate. They may fuse well ahead of the critical developmental stage or just barely make it in time, we have no way of telling. However, if they don't meet by the critical stage a cleft palate results. If they are close together at the critical stage, a small cleft will result, perhaps only a bifurcated uvula. If they are far apart, a more severe cleft will result. We call that critical difference in liability the threshold. Beyond the threshold, disease results. Below the threshold, normal development is observed. But the underlying liability is distributed as the normal curve shown in Figure 15.

DEGREE OF RELATIONSHIP AND GENES IN COMMON

Since one is not following a single locus with dominance or recessivity but is following several loci that act in concert, counseling for multifactorial inheritance diseases requires a different approach from that taken for Mendelian inheritance diseases. One has to calculate the number of genes in common. The easiest way to do that is to change the way we construct pedigrees. Instead of the familiar sibship method we use the pathway to common ancestor method. It is shown in Figure 16.

Figure 16. Conversion of a standard pedigree to a path coefficient pedigree for determining the fraction of genes in common.
In Figure 16, Pedigree A represents the standard method of pedigree construction. Pedigree B represents the pathway system of pedigree construction. It is much easier to see how genes flow from generation to generation in Pedigree B. In Figure 16, II-2 and II-3 are brother and sister. They have two common ancestors, I-1 and I-2. To determine the fraction of genes II-2 and II-3 have in common one simply counts all of the pathways and their connecting lines through the common ancestors. There is one line from II-2 to I-1, and a line from I-1 to II-3. That is one pathway with two lines of descent. There is another line from II-2 to I-2, and a line from I-2 to II-3. That is a second pathway with two lines of descent. These are the only pathways from II-2 to II-3. The fraction 1/2 is then raised to the power of the number of lines of descent and summed for each possible pathway, (1/2) 2 for the pathway through I-1, and (1/2)2 for the pathway through I-2, making a total of 1/2. Brothers and sisters have, on average, 1/2 of their genes in common.

A parent and offspring, say I-1 and II-2 also have 1/2 of their genes in common. There is only one pathway between them and only one line in that pathway, (1/2)1. Other relationships follow in the same manner. In Figure 16, III-1 and III3 are first cousins. There are two pathways connecting the two individuals, one through I-1 and the other through I-2, each with four lines. Their fraction of genes in common is then (1/2)4 + (1/2)4 or 1/8. First cousins have 1/8 of their genes in common. A grandparent and grandchild have 1/4 of their genes in common. There is a single pathway with two lines of descent. III-1 and IV1 are first cousins once removed. Again there are two pathways, one through I-1 and the other through I-2, each with 5 lines, (1/2)5 + (1/2)5 or 1/16 of their genes in common. The degree of relationship is often used rather than the fraction of genes in common. The degree of relationship is simply the power to which (1/2) is raised to reach the fraction of genes in common. First degree relatives have (1/2) of their genes in common. Second degree relatives have 1/4, (1/2) 2, of their genes in common, etc.

Figure 17. Method of calculating the recurrence risk of a multifactorial trait to first degree relatives.

If one returns to the normal curve for liability shown in Figure 15, one can now see where various relatives of affected lie in relationship to the threshold. Figure 17 shows these calculations for first degree relatives. The mean of affected can be calculated by dividing the affected rate by 2 and plotting that area under the normal curve. If a multifactorial disease affects 1/2000 offspring of normal x normal matings, then the mean of affected is an area of 1/4000. The number of standard deviations this area is from the mean can be found in statistical tables. Since first degree relatives have 1/2 their genes in common with their affected relative, first degree relatives of affected will be 1/2 of the way between the mean for affected and the population mean. This can also be calculated. A new normal curve with the same variance is then plotted using the mean for first degree relatives. The threshold does not move, so the overlap of the threshold will give the probability of recurrence to first degree relatives of affected. This is shown in Figure 18. Don't worry, you won't be asked to do this on an examination. This was done solely for the purpose of demonstrating the method for calculating the recurrence risk to relatives of affected. In practice you will simply look up the disease in the proper atlas and find the recurrence risks listed.

Figure 18. Recurrence risk to first degree relatives of affected individuals.

TWO THRESHOLD DISEASES


In many multifactorial diseases the two sexes have different probabilities of being affected. For example, pyloric stenosis occurs in about 1/200 newborn males but only in about 1/1000 newborn females. This means that there is a double threshold, one for females and one for males, with the female threshold farther from the mean than that for the male. However, since it takes more deleterious genes to create an affected female, she has more genes to pass on to the next generation. Her male offspring are at a relative high risk of being affected when compared to the population risk.

SEVERITY OF DISEASE AND RECURRENCE RISK


Unlike Mendelian traits with variable expressivity, where the recurrence risk is the same no matter how severely the individual is affected, multifactorial traits have a higher recurrence risk if the relative is more severely affected. In multifactorial traits, the more severely affected the individual, the more genes he/she has to transmit, and the higher the recurrence risk.

MULTIPLE AFFECTED OFFSPRING AND RECURRENCE RISK


Another difference is the presence of multiple affected individuals within a sibship. In Mendelian traits the number of affected in a family did not change the recurrence risks. But multiple affected children does change the recurrence risk for multifactorial traits. The presence of one affected child means the parents probably are midway between the mean for affected and the mean of the normal population, but the presence of a second affected child means they probably are closer to the threshold, and hence, have a higher recurrence risk should they choose to have another child.

CONSANGUINITY
Consanguinity also increases the probability of an affected child for a multifactorial trait, but only slightly when compared to rare autosomal recessive diseases. First cousin matings may increase the risk for two normal

individuals to have a child with a multifactorial disease by about two fold when compared to the risk for unrelated individuals.

HALLMARKS OF MULTIFACTORIAL INHERITANCE


In summary, the hallmarks for multifactorial inheritance are: 1. Most affected children have normal parents. This is true of diseases and quantitative traits. Most geniuses come from normal parents, most mentally challenged come from normal parents. 2. Recurrence risk increases with the number of affected children in a family. 3. Recurrence risk increases with severity of the defect. A more severely affected parent is more likely to produce an affected child. 4. Consanguinity slightly increases the risk for an affected child. 5. Risk of affected relatives falls off very quickly with the degree of relationship. Contrast this with autosomal dominant inheritance with invomplete penetrance, where the recurrence risk falls off proportionately with the degree of relationship. 6. If the two sexes have a different probability of being affected, the least likely sex, if affected, is the most likely sex to produce an affected offspring.

LINKAGE AND MAPPING

INTRODUCTION
hen the loci of two genes are on the same chromosome, they are said to be syntenic. When they are on the same chromosome, and are close enough together that they do not segregate independently, they are said to be linked. Linkage is a powerful tool in modern genetic counseling. In many families it allows predictions of an outcome rather than probabilities of an outcome. The human genome project now occupies a major effort of the human genetics community. Its first goal is to establish the precise chromosomal locus for each known Mendelian genetic trait, as well as any other "marker" loci. These "marker" loci may help to understand the inheritance of multifactorial traits, or they may be helpful in refining recurrence risk estimates in some families. This first goal is to be completed in two steps: first establish a genetic map and second convert the genetic map into a physical map of precise chromosomal position for each gene. This goal is supported by

a vast majority of human geneticists. The second, and more controversial goal of the human genome project is to establish a complete DNA sequence for each chromosome. Linkage studies in experimental organisms quickly led to the accumulation of rather complete maps of loci on individual chromosomes. The constraints of small family size, the lack of available polymorphic loci for normal variation, and random mating patterns has delayed the establishment of linkage maps in humans until new techniques beyond those of classical Mendelian genetics were developed. These techniques are now available. However, human linkage studies, no matter how complex, like those in experimental organisms are based upon the consequences of the chromosomal movement and recombination in meiosis. The student would do well to review the events of the first meiotic division and their consequences.

X-LINKAGE
Mendelian inheritance patterns are sufficiently powerful to establish the fact that a gene lies on the X chromosome. We discussed X-linked dominant and X-linked recessive inheritance in Mendelian genetics. It is not difficult to establish that two traits are syntenic, that is, both traits show X-linked inheritance. In developing chromosomal maps the genetic distance between the various loci is the first thing that needs to be established. If the two loci are far apart, so that a chiasma always occurs between them during first meiotic division, the genetic distance between them cannot be established even though they are syntenic. If, however, the two loci are sufficiently close together that recombination occurs, but not all the time, the genetic distance can be estimated. Figure 19 reviews the chromosomal movements and recombination of the two X chromosomes of the female in first meiotic division.

Figure 19. Recombination of hemophilia A and glucose 6-phosphate dehydrogenase deficiency during first meiotic division of oogenesis, showing the four possible resulting gametes.
In A, the female is a double heterozygote for hemophilia B and glucose 6phosphate dehydrogenase deficiency, two X-linked recessive traits. Only one pair of homologous chromosomes is shown. In B, the chromosomes have paired and two of the chromatids have exchanged parts, the result of a chiasma between the two loci. Only one chiasma is shown. At anaphase of first meiotic division the two centromeres will move to opposite poles. In C, the four resulting possible gametes of second meiotic division are shown. Gametes 2 and 3 are recombinants, gametes 1 and 4 are nonrecombinants, sometimes called parental types. Linkage distances are assigned on the basis that chiasma formation is random, that is, the farther apart the two loci, the more likely a chiasma will occur between them. With this assumption, which has proven to be approximately true, the ratio of recombinants to nonrecombinants gives an indication of the genetic distance between the two loci. If 5% of the gametes are like chromosome 2 and 5% are like chromosome 3, then there is 10% recombination. The two loci are said to be linked at a distance of 10 centimorgans. (1% recombination equals 1 centimorgan, named for T.H. Morgan, the first to discover linkage.) The theoretical maximum distance that can be measured in genetic studies is 50 centimorgans, or 50% recombination. At this distance all four of the gametic

products of a double heterozygote would have an equal chance of appearing, just as they would have if they were on different chromosomes with independent assortment. In actual practice, with the relatively small size of human families, greater distances than 25 centimorgans are extremely difficult to measure. When the two recessive alleles are on different homologs in the double heterozygote, as they are in Figure 19, linkage is said to be in repulsion, or trans. When they are on the same homologous chromosome in the double heterozygote, linkage is said to be in coupling, or cis. Unless the two loci are extremely close together, linkage in cis is equally likely as linkage in trans in the general population. There is no a priori reason to choose one over the other. X-linkage distances in a pedigree are most easily estimated when a grandfather expresses both recessive genes. Since he has only one X chromosome, linkage must be in coupling. His daughters' sons should be doubly affected or normal. Any grandson expressing only one trait must be a recombinant resulting from crossing over. A pedigree demonstrating the grandfather method of mapping is shown in Pedigree 10.

Pedigree 10. The grandfather method of estimating linkage distances for X-linked traits. I-1 has hemophilia B and is color blind.
In Pedigree 10, I-1 is a color blind hemophiliac. He has the recessive allele for each trait. His sons will all be normal, since they receive his Y chromosome and their mother's X chromosome. However, all of his daughters will be obligate carriers for both recessive alleles, hemophilia and color blindness. By chance, half of their sons should be normal and half should be doubly affected, if there is no recombination. I-1 has 8 grandsons through his daughters, two are normal, 5 are doubly affected, and one is color blind. III-4, the color blind child, is a recombinant. In this pedigree the best estimate of linkage distance is 12.5 centimorgans. Of course, several other families will have to be added to this sample for meaningfully accurate estimates to be made.

AUTOSOMAL LINKAGE
X-linkage is greatly simplified because there can be no recombination in the male. He has only one X chromosome and it is passed on intact to his daughters. In autosomal linkage studies, this does not hold. Recombination can, and does, occur in both oogenesis and spermatogenesis. Useful pedigrees for linkage studies using classical methods are quite limited. Other methods must be employed. These are discussed quite well in Gelehrter, Collins, and Ginsburg, 2nd ed., Chapters 9 and 10, and need not be replicated here. The student is referred to these chapters and is responsible for all of the material they contain. Sample questions from this material are included.

POPULATION GENETICS

INTRODUCTION
he applications of Mendelian genetics, chromosomal abnormalities, and multifactorial inheritance to medical practice are quite evident. Physicians work mostly with patients and families. However, as important as they may be, genes affect populations, and in the long run their effects in populations have a far more important impact on medicine than the relatively few families each physician may serve. It is important that certain polymorphisms are maintained so that the species may survive, even at the expense of individuals. Genetic polymorphisms often are detrimental to the homozygote, but they allow others of the species to survive. Before medical intervention was possible, populations that lacked the sickle cell anemia allele could not survive in the malaria regions of West Africa. Those that had the sickle cell anemia allele survived, and the gene remains in the population at

high frequency today, even though the homozygous recessive phenotype was at a severe disadvantage in the past. The high rate of thalassemia in people of Mediterranean origin, the high rate of sickle cell anemia in people of West African descent, the high rate of cystic fibrosis in people from Western Europe, and the high rate of Tay-Sachs disease in ethnic groups from Eastern Europe may all owe their origin to environmental factors that cause changes in gene frequencies in large populations by giving some advantage to heterozygotes who carry a deleterious allele. Although one may never use the calculations of population genetics in medical practice, the underlying principles should be understood. Population genetics is also the most widely misused area of human genetics, sometimes bordering on "vigilante genetics," a term coined by Newton Morton. Persons have mistakenly applied population genetics to "prove" race superiority for intelligence and aptitudes, and have misused it in eugenics. As an educated and, I hope, a respected member of your community you must be alert to "vigilante genetics." Population genetics is concerned with gene and genotype frequencies, the factors that tend to keep them constant, and the factors that tend to change them in populations. It is largely concerned with the study of polymorphisms. It directly impacts counseling, forensic medicine, and genetic screening.

GENE AND GENOTYPE FREQUENCIES


CODOMINANT ALLELES Consider a population of 1000 individuals all typed for the simplest test at the MN blood group locus. At its most simplistic form this locus can be reduced to a codominant system with two alleles M and N. (In reality it is considerably more complex than this but this simple form will suffice for our examples.) Every individual in the population will be either M (having two M alleles), MN (heterozygous), or N (having two N alleles). Suppose the blood typing results were as follows: 300 M individuals, 600MN individuals, and 100 N individuals. You probably want to ask, "What is the gene frequency of the M allele in the above population of 1000 individuals?" I'm glad you're interested! 1000 individuals each have two alleles at the MN locus = 2000 genes Each M individual has 2 M alleles 300 x 2 = 600 M alleles Each MN individual has 1 M allele 600 x 1 = 600 M alleles

There is a total of 1200 M genes in a population of 2000 genes. The gene frequency of the M allele is 1200/2000 = 0.6 I'll bet you want to know, "What is the gene frequency of the N allele?" Well, I'll show you how to find out. Each MN individual has 1 N allele 600 x 1 = 600 N genes Each N individual has 2 N genes 100 x 2 = 200 N genes Again, there is a total of 2000 genes in the population for the MN locus. The gene frequency of the N allele is 800/2000 = 0.4 Notice that when there are only two alleles in the population, their gene frequencies must add to 1. If they don't, you've done something wrong. This counting method of calculating the gene frequency must be used whenever the heterozygote can be detected. Gene frequency = (2 x homozygote + heterozygote) / 2 x population Gene frequency for one allele = 1 - gene frequency of the other allele These two general formulas assume nothing of the population, only that it is a single interbreeding group. All other methods make some assumptions of the population in order to simplify calculations. HARDY-WEINBERG EQUILIBRIUM For many human autosomal recessive traits the heterozygote cannot be distinguished from the normal homozygote. When this occurs the HardyWeinberg equilibrium is assumed to apply. These authors, Hardy in England and Weinberg in Germany, used different approaches but came to the same conclusions in 1908. They made several assumptions of the population: 1. 2. 3. 4. 5. 6. Large population Random mating No effect of recurrent mutation No selection against any phenotype No migration in or out of the population Autosomal locus

Under these assumptions, Hardy and Weinberg found that the gene frequency and the genotype frequency in the population do not change from generation to generation. Furthermore, if the frequency of the dominant allele A in the founding population was p , and the frequency of the recessive allele a in the founding population was q, then after one generation of random

mating the genotype frequencies would remain fixed and would be in the ratio:
p2 (AA) 2pq (Aa) q2 (aa) Since there are only two alleles in the population, p+q = 1 and p2 + 2pq + q2 = 1

If you want to see evidence that this is true, see Figure 20. If, on the other hand, you believe everything you read, and only want to study what will be covered on the examination, continue on.
Consider a population of two types of individuals, 50% AA and 50% aa (p andq = 0.5). Then with random mating one should have the following: Mother AA AA aa aa Father Frequency AA aa AA aa 0.5 x 0.5 0.5 x 0.5 0.5 x 0.5 0.5 x 0.5 Total AA 0.25 Offspring Aa 0.25 0.25 0.25 0.50 0.25 0.25 aa

Even though there were no heterozygotes in the founding population, after one generation of random mating, the genotype frequencies are in the ratio of p2(AA), 2pq (Aa), and q2 (aa), and the gene frequencies remain p = 0.5 and q = 0.5. Both remain fixed in those frequencies for future generations. Using a table similar to the above, the student can easily calculate the frequencies in the next generation.

Figure 20.
Hopefully, someone will ask the question, "Is there any evidence that the human population meets the requirements of Hardy-Weinberg equilibrium, or is this just a mental exercise?" Of course there is evidence! Consider the following: In my experience, one may use several criteria for selecting a person to mate with, but one usually doesn't select a mate based on blood types at the MN blood group locus. Therefore, we might assume that this locus would be a good test of random mating. All of the other Hardy-Weinberg criteria also seem to be met. Mutations at this autosomal locus are rare. We know of no selective advantage or disadvantage in the present environment. And migration wouldn't be much of a factor if we took the sample at one short interval of time. This locus should provide a good test. We have already seen that gene frequencies and genotype frequencies for this locus can be determined without using assumptions of Hardy-Weinberg

equilibrium. Let's see if a real population sample is distributed as p2 (M), 2pq (MN), q2 (N). In 1975, Race and Sanger reported the typing results from 1279 individuals in London. They were not collecting these data for the purpose of testing for Hardy-Weinberg equilibrium, so they could not be accused of typing individuals until a certain distribution was achieved, a question that has always remained about Mendel's original studies. Race and Sanger found 363 persons were M, 634 were MN, and 282 were N. Using our original method of calculating gene frequencies, the frequency of the M allele (p) would be: p = (2 x 363) + 634 / (2 x 1279) = 0.53167 The frequency of the N allele (q) would be: q = (2 x 282) + 634 / (2 x 1279) = 0.46833 If the population were in Hardy-Weinberg equilibrium, then the number of M individuals should be p2 x 1279, the number of MN individuals should be 2pqx 1279, and the number of N individuals should be q2 x 1279, or
Observed Expected M 363 361.54 MN 634 636.93 N 282 280.53

For the MN blood group locus there can be little doubt that the conditions for Hardy-Weinberg equilibrium are met in the human population, at least the population in London where the sample was taken. The observed frequencies closely approximate what would be expected if the population were in HardyWeinberg equilibrium. This gives us the assurance that we can use Hardy-Weinberg as a method when the heterozygote cannot be detected. An example of the use of the Hardy-Weinberg principle in medical genetics is given below. Suppose there is an autosomal recessive disease where the frequency of affected in the population is 1/10,000. If the population is in Hardy-Weinberg equilibrium, this frequency would equal q2. The gene frequency of the recessive allele (q) would then be the square root of q2, or the square root of 1/10,000 which equals 1/100. The carrier (heterozygote) frequency (2pq) is usually approximated as 2q since p (0.99) is so close to 1. The carrier frequency is then 1/50. For an autosomal recessive disease with a population frequency of 1/10,000, the carrier frequency is 1/50. Put another way, on average, as many as 3 or 4 first year medical students at UIC are carriers of such a disease.

From time to time, certain groups have suggested that the way to eliminate a deleterious disease from the population is to not allow affected individuals to mate. The above example should provide some evidence that this will have little effect on gene frequencies in the population. Although the frequency of the disease is only 1/10,000, (we should have one affected first year medical student at UIC every 50 years) the carrier frequency is 1/50 (we should have 3 or 4 carriers at UIC in every incoming class). These phenotypically normal carriers will keep the gene in the population. If, by chance, a student in the first year class has a sibling with an autosomal recessive disease that is present at birth, the student would have a 2/3 chance of being a carrier. If that student were to have a child with an unrelated partner selected at random from the general population, and the disease frequency in the general population is 1/10,000, the probability of their child being affected is: 2/3 (prob. of student being a carrier) x 1/50(prob. of a random individual being a carrier) x 1/4 (prob. of two carriers producing an affected child) = 2/3 x 1/50 x 1/4 = 1/300 Compare that to the probability that two unrelated individuals, with no history of the disease in their families would have an affected child, when the carrier frequency is 1/50: 1/50 x 1/50 x 1/4 = 1/10,000 Since it is a stated goal of medicine to do what is best for the patient, what happens to genes in populations when exceptions to Hardy-Weinberg occur?

RECURRENT MUTATION
Although mutation rates are usually very low, geneticists have long been concerned about environmental factors that will lead to even slight increases. There are two general types of mutation, a mutation that changes a gene that makes a functional product into a gene that makes a nonfunctional product (forward mutation) and a mutation that changes a gene that makes a nonfunctional product into a gene that makes a functional product (reverse mutation). Several events can lead to a forward mutation, base change, base insertion, base deletion, etc., but a reverse mutation must correct the specific change that produced the original forward mutation. For example if a single base deletion caused the original forward mutation, then that base must be reinserted in exactly the same place for a reverse mutation to occur. In general, forward mutations occur at a frequency that is at least 10 times that of reverse mutations. A method of estimating forward mutation rates is given in

Gelehrter, Collins, and Ginsburg, 2nd ed., Chapter 4. Students will be well advised to read this chapter carefully. If is the forward mutation rate from a functional to a nonfunctional allele, and is v the reverse mutation rate from a nonfunctional allele to a functional allele at the same locus, an equilibrium will be established between these two mutation rates that determines q, the gene frequency of the nonfunctional allele. At equilibrium, q = /(+v) If v is truly one tenth the frequency of , then we can assign the value 1 for v and 10 as the value for . The above equation reduces to qequil = 10/(10+1) or 10/11 =0.90909090909 Gene frequencies for nonfunctional alleles tend to increase in the population because of recurrent mutation. They will not entirely eliminate functional alleles but they tend to replace them, and can, if no other factors are involved, reach very high frequencies. As a possible human example of the effects of recurrent mutation consider the following. In the ABO blood group system, there are two functional alleles, Aand B. Alleles A and B control transferase enzymes that connect the proper sugar molecule (glucosamine or n-acetyl glucosamine) to a common precursor substance. Most likely, B was the result of a rare mutation of the A allele. O is a nonfunctional allele that recognizes no substrate, and no sugar molecule is transferred, leaving the precursor unchanged. In the ABO system, O is now the most frequent allele. If there is no selective advantage, O should continue to increase at the expense of A and B. The derivations of the equations used to calculate the effects of recurrent mutation are shown in Figure 21. Again, if you are interested only in studying for possible test questions, this material is not required.
Recurrent Mutation Assume a population of N individuals with two alleles at a locus, D with a frequency of pand d with a frequency of q. At generation 0 there will be 2Np D alleles , or 2N(1q) D alleles, and 2Nq d alleles. Assume D mutates to d at a frequency of and that d mutates to D at a frequency of v. Assume that is 10 times as frequent as v. Then at generation 1 the number of dalleles (2Nq1) would be: 2Nq1 = 2Nq (from gen. 0) + 2N (1-q) (mutations from D to d) - 2Nqv (mutations from dto D)

This reduces to: q1 = q + (1-q) - qv Or the change in q = q1 - q or the change in q = q + (1-q) - qv - q At equilibrium the change in q = 0, so at equilibrium 0 = q +(1-q) -qv -q, or, qv = (1q),or, qv = - q This reduces to q (at equilibrium) = /(+v)

Figure 21.
SELECTION AGAINST THE RECESSIVE PHENOTYPE One factor assumed in the discussion of recurrent mutation was that the nonfunctional allele and the functional allele have the same selective advantage. This may be true of the ABO blood group system, but it is not usually true of autosomal recessive diseases. The disease state, by definition, is always a deleterious phenotype. In autosomal recessive diseases the phenotype is almost always the result of nonfunctional alleles in the homozygous state. If left untreated the recessive phenotype for a disease would be less fit than the heterozygote or normal homozygote. How does selection against the homozygous recessive individual affect gene frequencies in the population? Fitness, to a geneticist, is not the same as fitness to a movie director or a sports columnist. Fitness is not measured by physical attributes, it is measured by the number of offspring produced in the next generation that survive and reproduce. In a hunting-gathering society, the most fit person may have been the near sighted male who could not go on the hunt because he would stumble and make too much noise. If he were left behind to gather fruit and berries with the women, he may have become the most fit person in the tribe. Grandchildren, great-grandchildren, etc., are the best measures of the fitness of an individual. This has alway been my favorite explanation of why so many of us are near sighted, and why society changed from hunting-gathering to agriculture. It's all population genetics! The most fit phenotype in the population is assigned a fitness of 1. If there are two equally fit phenotypes, each is assigned a fitness of 1. Those less fit must be assigned a fitness of less than 1. The difference between 1 and the fitness value is called the selection coefficient. The relationship between fitness, w, and the selection coefficient, s, is given by the equation, w = 1-s. The textbook uses f as the symbol for fitness, although historically most geneticists reserve f as the symbol for the inbreeding coefficient and use w as the symbol for fitness.

The effect of selection against the recessive phenotype is that, no matter how little the selection coefficient, as long as s is not 0, recessive alleles will be lost at each generation until no more remain in the population. Selection tends to reduce nonfunctional recessive alleles from the population; recurrent mutation tends to create nonfunctional recessive alleles in the population. The derivations of the effects of selection against the recessive phenotype are shown if Figure 22. Again, the material in Figure 22 will not be examined in this course.
Selection Against the Recessive Phenotype Generation 0 Fitness Generation 1 DD p2 1 p2 Dd 2pq 1 2pq dd q2 (1-s) (1-s)q2 Total 1 1-sq2

The frequency of q in generation 1, q1, = (2 x homozygote + heterozygote)/ 2 x total q1 = [2(1-s)q2 + 2pq]/ 2(1-sq2) , and q, the change in q, = q1 - q q = [(1-s)q2 + (1-q)q]/ (1-sq2) , which reduces to q = [-spq2]/ (1-sq2) q = 0 only when q = 0. There will be no equilibrium until the recessive allele is eliminated.

Figure 22.
BALANCE BETWEEN RECURRENT MUTATION AND SELECTION Since mutation tends to increase nonfunctional alleles in the population, and selection against the recessive phenotype tends to remove them, is there a point where these two will reach an equilibrium where gene frequencies remain stable from generation to generation? Again, if is the mutation rate, and s is the selection coefficient, an equilibrium will be reached when = sq2 If the fitness of the homozygous recessive individual is 0, that is, the individual with that phenotype cannot reproduce, then s equals 1 and the above equation reduces to = q2 The disease frequency cannot go lower than the recurrent mutation rate, even if affected individuals cannot reproduce. The derivations of these equations are shown in Figure 23.

Balance Between Selection Against the Recessive Phenotype and Recurrent Mutation For mutation, the change in q = - q -qv. For selection, the change in q = [-spq2]/ [1-sq2]. If they balance at an equilibrium, the net effect is that they should sum to 0. - q - qv + ([-spq2]/[1-sq2]) = 0 To simplify calculations, we will get rid of second order variables (qv) is only 1/10 of (q) and can be eliminated. Similarly, sq2 is very small in the denominator when compared to 1, and can be eliminated. This reduces the equation to -q - spq2 = 0 to first order magnitude. This reduces to - q = s(1-q) q2 or (1 - q) = (1-q)sq2 At equilibrium, = sq2 to first order magnitude.

Figure 23.
BALANCED POLYMORPHISM Some genes exist at a rather high frequency in the population because the heterozygote is more fit than either homozygote. The only documented example of this is sickle cell anemia in Western Africa. There are three major genotypes for the sickle cell locus, each producing a different phenotype, in West Africans, AA, or normal individuals, AS or heterozygote individuals (often called carriers), and SS individuals who will have sickle cell anemia. Without medical intervention, SS individuals will have a fitness less than 1. In the falciparum malarial environment of West Africa, AA and AS individuals get malaria, but AS individuals usually have much milder cases of the disease and usually survive while AA individuals are less likely to do so. The heterozygote is the most fit phenotype of the three. If the selection coefficient against the homozygous normal AA individual is t, and the selection coefficient against the homozygous SS individual is s, and if p is the frequency of the A allele and q the frequency of the S allele then an equilibrium will be reached in which p = s/(s + t) and q = t/(s + t). The gene frequencies at equilibrium are determined only by the relative sizes of the selection coefficients, not by their absolute magnitudes. The derivations of these formulas are shown in Figure 24. Again, you are not responsible for knowing how to derive these formulas.

Balanced Polymorphism Generation 0 Fitness Generation 1 DD p2 (1-t) p2(1-t) Dd 2pq 1 2pq dd q2 (1-s) q2(1-s) Total 1 1-tp2-sq2

The gene frequency of the q allele at generation 1, q1 = [2pq + 2q2(1s)]/2[1- tp2 - sq2] Again the change in q, q, = q1 - q and at equilibrium, q = 0 0 = [pq + (1-s)q2/ [1-tp2-sq2] Substituting (1- q) for p, this equation will reduce to: 0 = -spq + tp2 or sq = tp When (1-q) is substituted for p or (1-p) is substituted for q, this reduces to: q = t/(s + t) and p = q/(s + t).

Figure 24.
NON-RANDOM (ASSORTIVE) MATING Assortive mating in humans may occur to a limited degree for traits such as intelligence. In some studies, married couples have higher correlation coefficients for intelligence than do siblings. In modern western culture, we tend to marry someone who is about our own intelligence, although this is probably an over simplification. If intelligence were controlled by a single genetic locus with two alleles, S for smart and D for dumb, then three phenotypes would be possible, SSfor smart persons, SD for persons with average intelligence, and DD for persons who are mentally challenged. Of course, we know that intelligence is a multifactorial trait and not a single gene trait, but it is interesting to see what happens if it were a single gene trait with assortive mating where smart persons were only allowed to mate with smart persons, average persons with average persons, and mentally challenged only with mentally challenged. Strangely enough the gene frequencies do not change, only the genotype frequencies. The results are shown in Figure 25.
Assortive Mating Suppose a population started out as all heterozygotes, but heterozygotes mate only with other heterozygotes, homozygous dominant with homozygous dominant, and homozygous recessive with homozygous recessive. Then, over time, the following would result:

At each generation, half of the heterozygotes are lost with no change in gene frequency.

Figure 25.
Two different populations result, one smart, the other mentally challenged. Average gets lost. Assortive mating eventually results in two species being formed from one. SMALL POPULATIONS Gene frequencies in small isolate populations do not reflect those of the larger founding population from which they were derived because of two factors, founder effect and random genetic drift. Founder effect occurs when the population grew from a few founding individuals. A few individuals cannot represent all of the genomes of the founding population. As we discussed before, each of us is carrying from 1 to 8 mutant genes in the heterozygous state, even though we are normal. When the founding population is small, intermarriage must result even though steps are taken to avoid it. The mutations carried by the founders are in higher frequency than they would be in the general population from which the founders came. Island populations founded by pirates or shipwreck, that were isolated for several generations tend to have different gene and genotype frequencies because of founder effect. Similarly, religious isolates, where marriage outside the religion is forbidden, also have founder effects. Even if the founders of small isolate populations had exactly the same genotypes and gene frequencies of the original parent population, gene and genotype frequencies would change because of random genetic drift. Random genetic drift occurs because a small population cannot maintain randomness. Consider a population with 10 individuals with only two alleles at a locus, D with a frequency of 0.5 and d with a frequency of 0.5. By chance alone one would expect to find 10 D and 10 d gametes being passed to the next generation. But one may find 11 D and only 9 d gametes. The next generation, one could find 10 and 10 again, or could find 12 and 8. But suppose after drifting to 12 D and 8 d, by chance a really skewed sampling occurred and one got 15 D and 5 d. It would be difficult, if not impossible to

get back to the original 10 D and 10 d. Sampling errors in small populations are always going to occur if given enough opportunities. These errors assure that random genetic drift will always occur. Isolate populations never have the same gene and genotype frequencies as their founding populations. X-LINKED It is obvious that the major difference between autosomal loci and Xlinked loci in populations is that the males (usually half the population) have only one X. Males cannot have the distribution, p2, 2pq, and q2 because they have only one X, they have either the normal allele p, or the recessive allele, q. In males, gene and genotype frequencies are the same. Thus, the genotype frequencies in the male and female can never be the same. In addition, there can be no heterozygote x heterozygote mating class since there are no male heterozygotes, and as of this date females cannot mate and produce a child. X-linked traits can reach stable gene frequencies in males and females, but cannot reach Hardy-Weinberg equilibrium.