Acetylcholine

Paul M Salvaterra, Beckman Research Institute, Duarte, California, USA

Acetylcholine is a neurotransmitter found in the nervous systems of all animals. It is involved in the control of functions as diverse as locomotion, digestion, cardiac rate, fight and flight responses, secretion, learning and memory. Cholinergic dysfunction is associated with neuromuscular diseases such as myasthenia gravis and neurodegenerative disorders such as Alzheimer disease.

Introductory article
Article Contents
. Introduction . Major Neurotransmitter in Vertebrate Nervous System: Neuromuscular Junction, Ganglia, Brain . Major Neurotransmitter in Many Invertebrate Nervous Systems . Biosynthesis of Acetylcholine . Properties and Localization of Choline Acetyltransferase . Degradation of Acetylcholine . Properties and Localization of Acetylcholinesterase

Introduction
Studies of acetylcholine and cholinergic neurotransmission have played a key role in the development of nearly all aspects of our current understanding of chemical synaptic transmission. In the early part of the twentieth century, pioneering physiological and neurochemical experiments resulted in establishing the principle that release of neuroactive compounds, such as acetylcholine, on to eector cells or other neurons forms the basis of most types of intercellular communication. In these early studies, application of acetylcholine could mimic the eects of nerve stimulation on muscle contraction, the rate of heart beating, etc., and the compound was thus identied as the rst neurotransmitter substance. It was also noted that not all nerves released acetylcholine when stimulated, thus indicating specicity for the type of neurotransmitter substances present in particular neurons. Pharmacological work identied compounds, extracted primarily from plants, which dierentially blocked the action of acetylcholine on particular types of eector cells, leading to the concept of receptor specicity. The quantal nature of neurotransmitter release was also rst appreciated at cholinergic neuromuscular junctions. Finally, the nicotinic acetylcholine receptor was the rst ligand-gated ion channel to have its amino acid sequence established. Acetylcholine is a simple ester of the quaternary amino alcohol choline and acetic acid. Acetylcholine is positively charged at physiological pH, is freely soluble in water (usually supplied as a bromide or chloride salt) and is subject to rapid hydrolysis in solution by heat or alkali. Nuclear magnetic resonance studies indicate considerable exibility of the molecule in solution, and dierent conformations are thought to bind to dierent types of acetylcholine receptor. Acetylcholine functions primarily as a chemical neurotransmitter in the nervous systems of all animals. When a cholinergic neuron is excited, it releases transmitter into the synaptic cleft where it can bind to a number of dierent receptor proteins. The receptors for acetylcholine can be classied into two general categories based primarily on the actions of dierent plant alkaloids that aect their function: nicotinic (nicotine binding) or muscarinic (muscarine binding). Several dierent subtypes for each of these

. Packaging Acetylcholine into Synaptic Vesicles . Properties and Localization of the Vesicular Acetylcholine Transporter . Diseases Associated with Cholinergic Dysfunction . Acetylcholinesterase Inhibitors: Use as Insecticides, in Chemical Warfare and as Therapeutic Agents . Summary

general receptor classes have been characterized. The receptor binding event can be transduced into opening of cationic or anionic ion channels or coupled to some other metabolic signal such as phospholipid turnover rates or activation of second-messenger systems. Both inhibitory or, more commonly, excitatory responses are induced in the neurons or eector cells which receive the neurotransmitter signal, making acetylcholine-mediated neurotransmission particularly versatile. In addition to the ubiquitous presence of acetylcholine in the nervous systems of all animals, it is also found in a limited number of plants, bacteria, fungi and protozoa. This widespread distribution in a variety of species most likely indicates the appearance of acetylcholine-metabolizing proteins fairly early in evolutionary history. In vertebrates, acetylcholine is also found in non-neuronal tissues such as primate placenta and sperm where its functional role, if any, remains unknown.

Major Neurotransmitter in Vertebrate Nervous System: Neuromuscular Junction, Ganglia, Brain

Acetylcholine is found in, and used by, a wide variety of vertebrate neurons to carry out diverse functional roles. All vertebrate motor nuclei and spinal cord motor neurons make and use acetylcholine as a neurotransmitter for signalling skeletal muscles to contract. Motor activities such as locomotion, eye blinking, facial expressions, etc. are thus critically dependent on functioning cholinergic
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Acetylcholine

synaptic neurotransmission. Involuntary smooth muscle action also depends on acetylcholine neurotransmission. Activity such as diaphragm contraction (i.e. breathing) and gut contraction as well as excretory functions rely at least in part on cholinergic neurotransmission. Heart muscle responds to acetylcholine by slowing the rate of heart beating. Acetylcholine is also used as a transmitter in the preganglionic neurons of the sympathetic branch of the autonomic nervous system and for parasympathetic postganglionic neurons. Cholinergic neurotransmission thus plays an important role in regulating responses to stress or adverse environmental conditions, as well as maintaining internal homeostasis, in most animals. Temperature regulation through sweating and salivary secretion is regulated by acetylcholine. In the brain, acetylcholine-containing neurons are found to be broadly distributed with especially high concentrations in the basal forebrain nuclei (basal nucleus, the diagonal band of Broca and the medial septum), the caudate nucleus and prominent cholinergic inputs to the hippocampus. Cortical cholinergic neurons have been observed in certain species. While the specic functions of most central nervous system (CNS) cholinergic neurons are not known precisely, many pharmacological studies argue for a role in certain aspects of learning and memory or other higher-ordered thought processes. In addition to these primarily synaptic functions of acetylcholine, the transmitter may also play a role in blood pressure regulation by binding to nonsynaptic receptors in blood vessels or may control some aspects of early development by interacting with cholinergic receptors on fertilized eggs. The electric organs of Electrophorus and Torpedo are embryologically related to skeletal muscle tissue and are innervated primarily by a specialized CNS structure called the electric organ. The innervation is essentially purely cholinergic and, as a result, the brains of electric sh contain especially high levels of cholinergic macromolecules and acetylcholine relative to other types of vertebrates.

cholinergic neurotransmission in other species. Molluscs such as the sea slug Aplysia have a variety of identied cholinergic neurons in their CNS which have been studied extensively. At least three types of ionic channel-mediated responses have been characterized following activation of dierent types of acetylcholine receptors. In nematodes and annelids, acetylcholine can function as an excitatory neuromuscular transmitter in contrast to most other invertebrates which apparently use glutamate for this purpose.

Biosynthesis of Acetylcholine
Acetylcholine biosynthesis is accomplished by esterication of an activated acetyl group from coenzyme A with the tertiary amino alcohol choline. The reaction, shown in Figure 1, is catalysed by the enzyme acetyl-coenzyme Acholine-O-acetyltransferase (choline acetyltransferase) and in nervous system is thought to take place specically in cholinergic neurons. The acetyl-coenzyme A used for acetylcholine biosynthesis is derived from mitochondrial metabolism, whereas choline is derived from a variety of sources such as phospholipid turnover or reuptake of choline from the extracellular uid following breakdown of acetylcholine by hydrolysis. A specic Na 1 -dependent high-anity choline transporter is present on the plasma membranes of cholinergic neurons. Several studies have indicated that choline availability may be a rate-limiting step for acetylcholine production. It may even be possible to inuence the levels of acetylcholine by ingesting high levels of choline. One interesting facet of acetylcholine production is that the newly synthesized pool of neurotransmitter appears to be used preferentially for synaptic transmission, indicating possible coupling between transmitter biosynthesis and release.

Major Neurotransmitter in Many Invertebrate Nervous Systems

Acetylcholine is also used as a major neurotransmitter in a variety of invertebrate neurons with diverse functional roles. In most arthropods such as insects it is thought to be a primary sensory neurotransmitter for many types of peripheral neurons innervating a variety of chemosensory or mechanosensory specializations. Insects also contain relatively high concentrations of acetylcholine in their CNS, they have high levels of cholinergic macromolecules, and are particularly sensitive to application of anticholinergic compounds, which include a variety of organophosphate insecticides and alkaloids known to eect
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Properties and Localization of Choline Acetyltransferase

Choline acetyltransferase has been puried and characterized from a variety of species including Drosophila, nematodes, electric sh, rodents and humans. The enzyme is a single-subunit soluble globular protein with an Mr of approximately 68 000. Multiple isoelectric forms of choline acetyltransferase have been observed but their signicance is unknown and some may be generated artefactually

Choline acetyltransferase

Acetyl-CoA + choline

Acetylcholine + CoA-SH

Figure 1 Biosynthesis of acetylcholine is catalysed by choline acetyltransferase. CoA, coenzyme A.

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Acetylcholine

during purication by proteolysis or other posttranslational modications. The vast majority of choline acetyltransferase protein ( 4 95%) is found in an aqueous soluble form. A small proportion, however, appears to require detergent treatment for solubilization. It is not clear how the enzyme activity is associated with the particulate cellular fractions or what its functional signicance is, although it has been suggested to participate directly in ensuring that the transmitter is packaged into synaptic vesicles. Complementary deoxyribonucleic acid (cDNA) for choline acetyltransferase has been cloned from a variety of vertebrate and invertebrate species including Drosophila, Caenorhabditis elegans, porcine, rodent and human. In all species examined so far, there appears to be only a single gene for choline acetyltransferase termed Cha. In humans the genetic locus has been mapped to 10q11.2 by a variety of approaches. In rodents and humans choline acetyltransferase messenger ribonucleic acids (mRNAs) appear to be polymorphic as a result of a combination of alternative mRNA splicing and/or the use of alternative promoters to initiate transcription. In mice, for example, three dierent alternative 5 exons termed R, N and M appear to be alternatively spliced to a common second exon which contains the predicted codon for initiation of protein translation. At least two dierent forms of choline acetyltransferase mRNA have also been described in humans. The functional signicance of these alternative forms of mRNA is not known and only a single form of choline acetyltransferase mRNA has been observed in invertebrates. No vertebrate mutations in choline acetyltransferase have been described. Genetic mutations have been isolated and characterized in Drosophila and C. elegans. A number of lethal alleles are also available for the invertebrate Cha gene. Cha is an essential gene in Drosophila which shows late embryonic lethality in animals null for Cha function. Several temperature-sensitive conditional alleles have also been described for Drosophila Cha, which result in a number of interesting phenotypes such as paralysis, reduced motor activity and abnormal courtship behaviour. The nematode phenotype for hypomorphic Cha alleles is motor uncoordination. The localization of choline acetyltransferase has been accomplished by enzymatic assay of microdissected nervous system tissue as well as extensive immunocytochemical and in situ hybridization studies. Choline acetyltransferase is considered to be a reliable and specic marker for cholinergic neurons and the presence of the enzyme in a particular neuron is often taken as convincing evidence that that particular cell makes and uses acetylcholine as a neurotransmitter. Biochemical assays of choline acetyltransferase activity indicate that the highest levels are found in insect nervous system, the electromotor nucleus of electric sh and spinal cord ventral horn motor neurons.

In vertebrate nervous system immunocytochemical localization of choline acetyltransferase protein using a variety of monoclonal and polyclonal antibodies has revealed that the enzyme is present in all parts of cholinergic neurons. Positive reaction product has been observed in both symmetrical and asymmetrical synaptic buttons, with the major type being symmetrical. Certain groups of large projection neurons as well as local circuit neurons appear to be cholinergic. In spinal cord large a motor neurons as well as smaller g motor neurons, preganglionic projections of the autonomic nervous system, all stain with anticholine acetyltransferase antibody. In the brain choline acetyltransferase-positive neurons and synaptic buttons are widely distributed and include both local circuit as well as projection neurons. The cholinergic projection neurons of the midbrain and forebrain have been divided into eight groups (Ch1Ch8) based on their locations and projection patterns. They are often interspersed with choline acetyltransferase-negative neurons. Prominent cholinergic neurons have been found in the medial septum, the vertical and horizontal limbs of the diagonal band, the nucleus basalis, the postmesencephalic reticular formation, the medial habenula, the parabigeminal nucleus and the motor neurons of cranial nerve nuclei. Terminal elds containing choline acetyltransferase-positive synaptic buttons have been described in the thalamus, interpeduncular nucleus, superior colliculus, hippocampus and cerebral cortex. Regions of the CNS containing choline acetyltransferase-positive intrinsic neurons include the spinal cord, cerebral cortex, amygdaloid complex, neostriatum, ventral striatum, olfactory bulb, retina and hypothalamus. In invertebrates, such as Drosophila, choline acetyltransferase-positive neurons have been localized in many regions of the central and peripheral nervous system, and in general the localization is consistent with functions related to acquisition or processing of primary sensory information. Surprisingly, mutant Drosophila which have a wild-type cDNA transgene as their only source of choline acetyltransferase appear to have fairly normal behavioural functions, even when the pattern of transgene expression is quite dierent from the normal choline acetyltransferase expression pattern.

Degradation of Acetylcholine
Acetylcholine action is terminated by hydrolysis of the transmitter into free choline and acetate. The reaction at synapses is catalysed by the enzyme acetylcholinesterase, as shown in Figure 2. Other esterases that hydrolyse acetylcholine, such as butyryl or pseudocholinesterases, are also present at nonsynaptic sites, but their contribution to transmitter inactivation in vivo is unknown. Inactivation of transmitter by hydrolysis is rather unusual for small3

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Acetylcholine

Acetylcholinesterase

Acetylcholine

Choline + acetate

Figure 2 Hydrolysis of acetylcholine is catalysed by acetylcholinesterase.

molecule neurotransmitters since the actions of most others are terminated by removal of transmitter from the synaptic cleft by specic high-anity uptake systems located in the plasma membrane of releasing neurons and/or nearby cells. Most cholinergic neurons do, however, contain a specic plasma membrane Na 1 -dependent high-anity choline uptake system, which is believed to recover approximately 50% of the choline used for acetylcholine synthesis.

Properties and Localization of Acetylcholinesterase

Acetylcholinesterase exists in a number of molecular forms which appear to convert the enzyme between soluble and insoluble forms, but have no major eect on substrate specicity or catalytic activity. One class of molecular forms is made up of either monomers or homologous combinations of dimeric or tetrameric subunits. Varying degrees of posttranslational modication aect the hydrophobicity by adding a glycophospholipid to the C-terminal carboxyl group of the protein(s). The lipid allows the enzyme to be tethered to plasma membranes. The second major class of acetylcholinesterases is made up of combinations of heterologous subunits. These are composed of up to 12 catalytic subunits linked by disulde bonds to collagen tail-containing subunits, giving the nal protein assembly a pronounced axial asymmetry. The collagenous tail of asymmetrical acetylcholinesterase is thought to localize this form of the enzyme to the basil lamina in synaptic clefts of the neuromuscular junction. Another type of acetylcholinesterase found in brain has a phospholipid tail-containing subunit which facilitates association with membranes. Acetylcholinesterase has no obvious membrane-spanning domains and the protein is usually synthesized as a secreted protein with a hydrophobic leader sequence. The various forms of the enzyme have been puried from a variety of vertebrate and invertebrate species, and cDNAs have been isolated and characterized from several of these species. The dierent molecular forms arise via alternative splicing of a single gene transcript. Exons 1 and 2 are identical for symmetrical and asymmetrical forms of the protein. Alternative splicing to two dierent third exons gives rise to distinct C-terminal proteins. The specic attachment points for posttranslational modications and heterologous subunit assembly are contained within the divergent third exon sequences. The three-dimensional structure of acetylcholinesterase isolated from Torpedo
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californica has been determined. The catalytic mechanism of acetylcholinesterase is analogous to that of the serine proteases, even though there is no obvious primary structural homology between the cholinesterases and proteases. Several lines of evidence have mapped the human acetylcholinesterase gene to 7q22, while the pseudocholinesterase gene is at 3q25.2. Other genetic studies have established that acetylcholinesterase carries the YT blood group antigenic determinant. Genetic studies in avians have established that the multiple forms of acetylcholinesterase are coded for by a single genetic locus. In Drosophila several lethal and conditional alleles of the single acetylcholinesterase locus have been described. Complete absence of the enzyme results in late embryo lethality. Temperature-sensitive mutants exhibit a number of phenotypes which are similar to temperature-sensitive choline acetyltransferase alleles. Nematodes have two dierent acetylcholinesterase genes, ace-1 and ace-2. The distribution of acetylcholinesterase is signicantly broader than that of choline acetyltransferase, making it dicult to use immunocytochemical localization of this protein as a specic marker for cholinergic neurons. The enzyme is found in nearly all regions of the nervous system that exhibit choline acetyltransferase activity and in many places that do not. One interesting aspect of acetylcholinesterase concerns its localization at neuromuscular junctions. The greatest concentration of protein appears to be situated at the tips of junctional folds. Transmitter would thus be exposed to degradative enzyme activity, which could inactivate it before binding to cholinergic receptors located within the junctional folds. Apparently, the acetylcholinesterase is rapidly saturated with substrate during release of substantial quantities of transmitter from activated neuromuscular junctions.

Packaging Acetylcholine into Synaptic Vesicles

Acetylcholine is released at cholinergic synaptic sites in response to invasion of a depolarizing current into the presynaptic nerve terminal. The release process is Ca2 1 dependent and appears to involve synaptic vesicle fusion with the presynaptic plasma membrane. Spontaneous release of transmitter also occurs and has been statistically characterized as quantal in nature. The physical picture of a quantal unit of acetylcholine is thought to be the contents of a single synaptic vesicle. Since synaptic vesicles are ordinarily impermeable to acetylcholine, they depend on the action of a specic vesicular transport protein to move acetylcholine from its site of synthesis in the cytoplasm into the internal lumen of synaptic vesicles. The protein that accomplishes this process is called the vesicular acetylcholine transporter. The transporter functions as

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Acetylcholine

an acetylcholineH 1 antiporter by exchanging protons within the vesicle with cytoplasmic acetylcholine. The ratio of the acetylcholine concentration inside a vesicle relative to the cytosol has been estimated to be about 100 : 1. The high concentration of H 1 within vesicles, which serves as the driving force for the transporter, is established by an ionmotive adenosine triphosphatase.

The conservation and organization of these two distinct but related genetic functions may be related to coordinate control of expression at the transcriptional level. The cholinergic gene locus, shown schematically in Figure 3, thus has properties of a eukaryotic operon, ensuring that both choline acetyltransferase and the vesicular acetylcholine transporter are expressed in the same cells at the same time.

Properties and Localization of the Vesicular Acetylcholine Transporter

The vesicular acetylcholine transporter belongs to a fairly large gene family composed of a number of transporters which contain a general structure of 12 transmembranespanning domains. Other members of this family include the vesicular monoamine neurotransmitter transporters and a variety of plasma membrane neurotransmitter transporters. More distant relatives include sugar transporters and some bacterial drug resistance genes. The size of the vesicular acetylcholine transporter is approximately 60 kDa. The protein contains hydrophilic amino acid residues at the N and C termini and a large potentially glycosylated luminal loop between the rst and second transmembrane domains. The activity of the vesicular acetylcholine transporter can be blocked specically by the drug vesamicol in isolated intact synaptic vesicles. One of the major surprises in cholinergic neurobiology was the discovery of the genomic relationship between choline acetyltransferase and the vesicular acetylcholine transporter. Both genetic functions are arranged as a gene complex in all species so far examined (i.e. nematodes, Drosophila, rats, mice and humans). In nematodes and Drosophila, the coding regions of the vesicular acetylcholine transporter are nested entirely within the rst intron of the choline acetyltransferase gene and a common sequence is contained in each specic transcript. The specic transcripts are thought to arise by posttranscriptional RNA processing. Vertebrates appear to have specic transcripts with both shared and unique sequences. The latter are thought to arise from alternative promoter usage.

Diseases Associated with Cholinergic Dysfunction

The best understood human diseases of cholinergic dysfunction are disorders of skeletal neuromuscular transmission. Myasthenia gravis is characterized by episodic weakness in muscles primarily innervated by the cholinergic cranial motor neurons. The disease is caused by an autoimmune attack on postsynaptic cholinergic receptors, thus resulting in an eective block of neurotransmission. Current treatments include use of acetylcholinesterase inhibitors or procedures to reduce the level of antireceptor antibodies. Amyotrophic lateral sclerosis is characterized by degenerating spinal cord anterior horn cell motor neurons, leading to progressive muscular weakness and eventual atrophy. A familial form of this condition has recently been proposed to be due to defects in the superoxide dismutase gene. EatonLambert syndrome, a presynaptic neuromuscular disorder, results from impaired acetylcholine release at nerve terminals. Several CNS neurodegenerative diseases also have cholinergic dysfunction. Alzheimer disease is a common age-related progressive degenerative disorder of the CNS that results in impaired thinking, memory and behaviour. The number of cholinergic neurons in the basal forebrain (basal nucleus, diagonal band of Broca and medial septum) is substantially lower in patients with Alzheimer disease and the loss of these neurons may be related to the impaired cognitive functions. Several other types of CNS neurodegenerative disorders, such as Parkinson disease and Huntington disease, have aspects of cholinergic dysfunction. Although cholinergic neurons are not the primary type of degenerating cells in these diseases, the cholinergic symptoms are believed to result from an imbalance in cholinergic neurotransmission thought to be caused by a loss of inhibitory control. Although not technically a disease, smoking is a primary public health problem with a strong cholinergic component. Nicotine in tobacco smoke is believed to be an addictive stimulus for certain types of central cholinergic receptors, which reinforces the pleasurable aspects of smoking in spite of the obvious health-threatening consequences. Other cholinergic-related health problems involve the deliberate or accidental exposure to
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Common regulatory region

Transferase-specific exons

Shared exon

Transporter-specific exon

Figure 3 Organization of the cholinergic gene locus. The vesicular transporter and choline acetyltransferase share a common exon and transcriptional regulatory region.

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anticholinergic drugs or chemicals primarily used as insecticides.

Acetylcholinesterase Inhibitors: Use as Insecticides, in Chemical Warfare and as Therapeutic Agents

Inhibitors of acetylcholinesterase activity are often termed anticholinergics and include a variety of compounds that reversibly or irreversibly inhibit enzyme activity. Some representative structures are shown in Figure 4. Since acetylcholine is ordinarily removed rapidly from cholinergic synapses by esterase hydrolysis, the actions of these inhibitors can all be related to the persistence of neurotransmitter at the various eector organs, neuromuscular junctions or in the CNS. The action of most naturally occurring plant alkaloid inhibitors of acetylcholinesterase is easily reversed by high concentrations of acetylcholine; these inhibitors include such prototypical structures as physostigmine (eserine) or neostigmine which are loosely bound to the active site of the enzyme. Another class of inhibitors includes the highly toxic irreversible organophosphates such as diisopropyl uorophosphate. These irreversible inhibitors were rst used as eective insecticides (i.e. parathion and malathion) and are thought to work essentially by preventing termination of cholinergic neurotransmission (i.e. continual stimulation of cholinergic receptors). Certain members of this class of compounds have also been developed as chemical warfare agents (i.e. Tabun, Sarin, Soman and VX). The organophosphate inhibitors irreversibly phosphorylate a serine residue within

the active site of acetylcholinesterase that is essential for enzyme activity. The more toxic members of the organophosphates class of inhibitors undergo secondary chemical reactions when covalently attached to acetylcholinesterase, termed ageing. Aged enzyme is impossible to reactivate and thus recovery from organophosphate poisoning often requires removal of the organophosphate and synthesis of new enzyme. Symptoms of acute organophosphate poisoning can include excessive salivation and lacrimation, digestive system disturbances, muscle fasciculation and weakness, constriction of the pupils and depressed levels of consciousness or seizures. Treatment involves respiratory management and administration of atropine, which acts primarily by blocking cholinergic transmission at muscarinic type acetylcholine receptors. Therapeutic uses of acetylcholine esterase inhibitors include treatment for certain ocular conditions (i.e. to reduce intraocular pressure), enhancement of gastric contractions and/or intestinal motility, and enhancement of skeletal neuromuscular transmission (i.e. in the treatment of hypofunction of cholinergic neuromuscular junctions in conditions such as myasthenia gravis). More recent experimental studies are employing reversible inhibitors that cross the bloodbrain barrier to enhance central cholinergic transmission in Alzheimer disease.

Summary
Acetylcholine is a versatile neurotransmitter substance in the nervous systems of all animals. The actions of acetylcholine can be both inhibitory and excitatory, and examples of cholinergic synaptic transmission are seen in both the central and peripheral nervous system. The gene products that synthesize (choline acetyltransferase), package (the vesicular acetylcholine transporter), inactivate (acetylcholinesterase) and receive (muscarinic and nicotinic receptors) cholinergic signals have all been cloned and characterized in a variety of dierent species. Precise molecular information is not yet available for the plasma membrane choline transporter, which also participates in the cholinergic cycle. A number of human diseases are known which have various presynaptic and/or postsynaptic defects in cholinergic neurotransmission. Acetylcholinesterase is a primary target of clinically useful drugs as well as toxic insecticides and chemical warfare agents. Drugs that block the action of acetylcholine at receptors are also used clinically and several toxins have similar actions.

Reversible inhibitors
CH3 O C O N CH3 N CH3 N H CH3 CH3 CH3 N CH3 O O C N CH3 CH3

Physostigmine Irreversible inhibitors

CH3 CH3 CH3 CH CH CH3 O P O F O H3 H3 CH2 P CH2 O NO2 S

Neostigmine

CH3 CH CH3 O P CH3 F O

Diisopropyl fluorophosphate

Parathion [O,O-diethyl O-(4nitrophenyl)-phosphate]; insecticide

Sarin (isopropyl methylphosphonofluoridate); nerve gas

Further Reading
Bardin PG, van Eeden SF, Moolman JA, Foden AP and Joubert JR (1994) Organophosphate and carbamate poisoning. Archives of Internal Medicine 154: 14331441.

Figure 4 Some common types of acetylcholinesterase inhibitors.

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Birks RI and MacIntosh FC (1957) Acetylcholine metabolism at nerveendings. British Medical Bulletin 13: 157161. Dale HH (1954) The beginnings and the prospects of neurohumoral transmission. Pharmacological Reviews 6: 713. Parsons SM, Prior C and Marshall IG (1993) Acetylcholine transport, storage, and release. International Review of Neurobiology 35: 279 390. Salvaterra PM and Vaughn JE (1989) Regulation of choline acetyltransferase. International Review of Neurobiology 31: 81143. Taylor P (1996) Anticholinesterase agents. In: Hardman JG, Limbird LE, Molino PB, Ruddon RW and Goodman AG (eds) Goodman &

Gillmans The Pharmacological Basis of Therapeutics, 9th edn, pp. 161176. New York: McGraw-Hill. Taylor P (1996) Agents acting at the neuromuscular junction and autonomic ganglia. In: Hardman JG, Limbird LE, Molino PB, Ruddon RW and Goodman AG (eds) Goodman & Gillmans The Pharmacological Basis of Therapeutics, 9th edn, pp. 177197. New York: McGraw-Hill. Usdin TB, Eiden LE, Bonner TI and Erickson JD (1995) Molecular biology of the vesicular ACh transporter. Trends in Neurosciences 18: 218224. Wu D and Hersh LB (1994) Choline acetyltransferase: celebrating its ftieth year. Journal of Neurochemistry 62: 16531663.

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