PH 121: GROSS & MICROSCOPIC ANATOMY

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CONNECTIVE TISSUE
Dr. Lorenz Alvarez | August 20, 2018

OUTLINE
I. Connective Tissue III. Classification of
II. Components of Connective Tissues
Connective Tissue IV. Laboratory Slides
A. Ground Substance V. References
B. Extracellular Fibers
C. Cells

I. CONNECTIVE TISSUE
 Connective Tissue consists of widely separated cells in an
abundant extracellular matrix
 One of the four major types of tissues
 Epithelial tissue
 Connective tissue
 Muscular tissue
 Nervous tissue Figure 1. Major Glycosaminoglycans of Connective Tissue
 Serves as the medium of diffusion
 Origin: 2. PROTEOGLYCANS
 Mesenchyme derived from mesodermal cells (except for the  Consist of a core protein to which are covalently attached
connective tissue in the nervous system which is derived from to various numbers and combinations of the GAGs (except
the ectoderm) hyaluronic acid)
 Mesenchyme is formed by invading mesodermal cells that  Like glycoproteins, they are synthesized on RER, mature in
surround developing organs the Golgi apparatus, where the GAG side-chains are
 Functions: added, and secreted from cells by exocytosis.
1. STRUCTURAL - functions as tissue glue, covering (capsule  Unlike glycoproteins, proteoglycans have attached GAGs
of organs), and framework which often comprise a greater mass than the polypeptide
 Stroma – supporting tissues found in glands core
 Parenchyma – secretory/ active portion of glands  Examples of Proteoglycans:
2. DEFENSE – has immunologic properties  PERLECAN: key proteoglycan in basal lamina
3. NUTRITION – metabolic support  AGGRECAN:
 Fat tissues often provide the metabolic support in o Having a core protein heavily bound with chondroitin
connective tissues and keratan sulfate chains. A link protein joins
 H2O – most abundant in the body; enables substances to aggrecan to hyaluronan. Abundant in cartilage,
diffuse in the body aggrecan-hyaluronan complexes the space between
collagen fibers and cells and contribute greatly to the
physical properties of this tissue
II. COMPONENTS OF CONNECTIVE TISSUE  DECORIN: with very few GAG side chains that binds the
surface of type I collagen fibrils
A. Ground Substance  SYNDECAN: an integral membrane core protein
 Gel-like matrix: occupying space between the cells and fibers of providing an additional attachment of ECM to cell
connective tissue membranes.
 Highly hydrated, transparent, and complex
 Abundance of ground substance varies on type of tissue 3. MULTIADHESIVE GLYCOPROTEINS. (Mescher, 2013)
 Viscous and amorphous is due to much bound water (most  Have more protein than carbohydrates
important part of ground substance)  More of communicational in function
- Accountable for the properties of connective tissue  FIBRONECTIN
 Stains with detection of carbohydrates o Dimer (made of 2 molecules); has binding domains
 INTERSTITIAL FLUID - the water in the connective tissue and for cells, collagen fibers, and glycosaminoglycans
has ion composition similar to blood plasma o These domains are for connecting cells to the fibrous
 Mixture of three major kinds of macromolecules: and amorphous components of the connective tissue
1. GLYCOSAMINOGLYCANS (GAGs) o Fibronectin substrate provides specific binding sites
 Long polymers of repeating disaccharide units, usually for integrins (transmembrane receptors that facilitate
hexosamine and uronic acid. cell-extracellular matrix adhesion) and is important
 The major glycosaminoglycans of connective tissue are: both for cell adhesion and cellular migration through
 Chondroitin-4-sulphate the ECM
 Chondroitin-6-sulphate  LAMININ
 Dermatan sulphate o Trimer; cross-shaped
 Keratan sulphate o Type IV collagen and specific proteoglycans,
 Heparin sulphate o Provides adhesion to cell membranes, collagen, and
 Heparan sulphate some proteoglycans;
 HYALURONIC ACID o With binding sites for integrins
o Not sulfated; only GAG that is not associated with o All basal and external laminae are rich in laminin,
proteins which is essential for the assembly and maintenance
o Largest and most ubiquitous GAGs; a very long of these structures
polymer of the disaccharide glucosamine-  THROMBOSPONDIN
glucuronate o Binds to cells, collagen fibers and fibronectins
o Substance found in blood
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  FIBRILLIN
o Essential for the formation of elastic fibers
B. EXTRACELLULAR FIBERS
 Extracellular fibers are made of proteins
 There are three types of fibers:
1. COLLAGEN FIBERS
 Collagen
 Most abundant protein synthesized by fibroblast
 Procollagen  microfibrils (tropocollagen)  fibrils 
collagen fiber
 3 peptide chains: glycine (33.5%), proline (12%),
hydroxyproline (10%)
 Collagen is trihelical in structure
 Responsible for the firmness of connective tissue
 Has many cross striations
 Heterogenous - many different kinds of collagen 24
types
 Has 4 focal main types:
 TYPE 1: Cross striated fibrils; found in skin, bone, and
tendon and capsules of organs
 TYPE 2: very thin fibers; found in hyaline and elastic
cartilage, intervertebral disk and vitreous body of the
eye
 TYPE 3: slender fibers, reticular fibers; found in the
muscles, spleen, kidney and uterus, and the wall of
blood vessels
 TYPE 4: Under electron microscope, found in the basal
lamina, Forms close network between heparin sulfate
for epithelium
* Types 1, 2 and 3 interstitial collagen are visible under
the light microscope
 Stains for collagen:
 Hematoxylin and Eosin – pink (eosinophilic bundles)
 Mallory’s trichome stain – blue
 Masson’s stain – green
 Sirius red – red
 1-20 μ in diameter, each made up of fibrils (75 nm in
diameter)
 Makes up 30% of dry body weight; predominant fibrous
component of connective tissues
 Non-branching, elongated, tortuous, and cylindrical
 Abundant in bone, dermis, and cartilage
 Extremely strong and resistant to forces
Figure 2. Three Stages of Elastic Tissue Formation” Leftmost: Oxytalan, Center:
3. RETICULAR FIBERS
Figure 2. Collagen Fibers
2. ELASTIC FIBERS
 Made of elastin protein allows the connective tissue to
stretch
 0.2-1.0μ in diameter; made up of fibrils 10nm in diameter
 Do not branch but form sheets/layers/laminae
 Stains:
o Dark blue/Black with Orcein stain
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 Connective tissue can stretch only so much before its
thick, ropelike collagen fibers become taut. Then, when
the tension lets up, elastic fibers snap the connective
tissue back to its normal length and shape. (Marieb &
Hoehn, 2013)
 Can expand up to 200% its original length and return to
original size
 Abundant in large blood vessels (especially the aorta) or
tissues which need elastic properties (such as the lungs,
bladder)
 Shape can be flat and stellate, fusiform cells
 FIBROBLASTS - dominant cell type of the connective
tissue, responsible for the production of fibrous tissues
 Stages of elastic tissue formation:
1. OXYTALAN
 First stage in the formation of elastic
fibers, large concentrations are found
in the ligaments and gingivae
 A developing fiber consists of many
10nm in diameter fibrillin microfibrils
composed of molecular subunits
secreted by fibroblasts.
2. ELAUNIN
 Elastin proteins are found
interspersed in oxytalan fibers, they
look like globs with microfilaments
 Elastin is deposited on the scaffold of
microfibrils, forming and growing into
amorphous structures.
3. ELASTIC FIBER
 Long curly filaments filaments has
Desmosine
 Elastin accumulates and ultimately
occupies the electron-dense center of
the elastic fiber
 Elastin, the protein of elastic fibers, contains 2 unusual amino
acids:
 DESMOSINE
 ISODESMOSINE
 responsible for the elastic recoil of the elastic fibers.
Elaunin, Rightmost: Mature Elastic Fiber
 Type III Collagen + associated glycoproteins
 0.5-2.0μ in diameter; made up of fibrils 35nm in diameter
 Continuous with collagen fibers
 Form an extensive branching network
 Stains:
 Black with Silver stain (argyrophilic)
 Red/orange with Periodic Acid-Schiff (PAS)
 Due to high content of sugar chains bound to type III
collagen
 Abundant in smooth muscles covering the spleen and in the
liver, lymphoid
 Also form a delicate framework for endocrine glands, lymph
nodes, and the bone marrow
 Found in the basement of the epithelial tissue
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Figure 3. Reticular Fibers in the Adrenal Cortex
C. Cells
I. FIXED CELLS
A. FIBROBLASTS
- Produces matrix fusiform cells rarely divide within normal
connective tissues
- Most abundant and ubiquitous in the CT
- Fibroblasts produce the matrix of CT
- Fusiform/spindle-shaped (with tapered ends)
- Deployed along collagen fibers (fibroblasts also produce
collagen fibers)
- ‘Sessile’: they rarely divide in normal CT, they are only
activated and divided when there is an injury in the CT
for rebuilding of matrix
- Secreted ECM components undergo further modification
outside the cell
B. ADIPOSE CELLS
- nucleus flattened to one side
- found in blood vessels
- Accumulates lipid in the cytoplasm thus, nucleus flattens to
one side
- Modified support cells specialized in the storage and
metabolism of fat (Young, et al., 2014)
- Collectively form adipose tissues
- Signet ring appearance
- Lipid is removed during slide preparation so fat cells appear
empty when viewed in LM
C. MESENCHYMAL CELLS
- Multipotent; can produce more than one type of
specialized cell of the body, but not all (Nogués & Creane, 2015)
- Fusiform or stellate; hard to distinguish from fibroblasts
- Undifferentiated and have large nuclei, with prominent
nucleoli and fine chromatin(Mescher, 2013)
II. FREE CELLS
 Mostly blood cells
 Motile cells, basically migratory
1. Leukocytes are transient cells of CT for they originate in bone
marrow, stay in blood and move to CT when only needed
2. Plasma and mast cells originate in bone marrow then reside
in CT where they function (Mescher, 2009)
A. MONOCYTES
- Round agranular cell with diameter of 9-17 μ
- Foamy grayish blue cytoplasm with eccentric kidney/bean-
shaped nucleus
- Precursor of macrophage
- Type of white blood cells
- Has a bean-shaped nucleus
- Produced in the bone marrows
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B. LYMPHOCYTES
- Spherical agranular cell with diameter of 7-9 μ
- Thin rim/halo of cytoplasm encircling a prominent round
nucleus
o Large intensely staining nucleus
- High count in viral infections
- Matures into:
o Killer T-cell
o B-cell (antibody production, parent of plasma
cells)
o Natural Killer (NK) cells
C. PLASMA CELLS
 Ovoid cell with eccentric round nucleus, basophilic
cytoplasm due to numerous RER and easily seen negative
Golgi apparatus
 “Clock-face appearance” as its distinct feature is due to
peripheral clumps of heterochromatin
 Arises from B-lymphocyte after it is ‘selected’ by an
antigen
 Larger than lymphocytes due to more abundant cytoplasm
 Secretes antibodies for immune response
D. EOSINOPHILS
 Round cell with bilobed nucleus and pink coarse
cytoplasmic granules that do not obscure the nucleus
(opposite condition in basophil)
 Migrates to sites of allergic reactions and parasitic
infections but does not phagocytose
 Granules contain hydrolytic enzymes as well as aryl
sulfatase and histaminase for controlling allergic
responses
 Few if normal, increased in mentioned circumstances
E. MAST CELLS
 Largest of free cells
 Typically oval (sometimes irregularly-shaped) cell with
nucleus centrally located (Mescher, 2013)
 Possesses abundant cytoplasmic granules (mast = stuffed
full of granules) that contain heparin, histamine, proteases
and cytokines
 Responsible for inflammatory response and severe
allergic reactions
 Has high content of acidic radicals in sulfated GAGs
leading to Metachromasia (Mescher, 2013)
o change in color of the stain due to dye
aggregation
o ex. Toluidine Blue (blue to purplish red),
Methylene Blue (blue to red-purple)
F. NEUTROPHILS
 Nucleus commonly has 3-4 lobes
 Has granules that are fine and staining faintly because
the granules do not attract either acidic or basic stain for
those are neutrophilic (neutral-loving) (Tortora, 2010)
 Not seen in normal connective tissue but only found
during an inflammation when it invades
 A phagocytic cell and the most abundant leukocyte
 Raised count is especially seen in bacterial infections
G. BASOPHILS
 release histamine and heparin
 Make up 0.5-1% of white blood cells
_____________________________________________________
*MACROPHAGES
Cannot be confined under either Fixed or Free cells
-shaped nucleus and
cytoplasm having irregular surface with protrusions
and pleats
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 Abundant lysosomes, vacuoles and debris in cytoplasm  Variation in: (i) predominance of cell type (ii)
Acts as phagocytic cell and secretory cell (releases prominence of fiber (iii) consistency
cytokines that activate and specify lymphocytes) a. MUCOUS
 Gives positive test for mucin
 Has mesenchymal CT
 RESIDENT
o Endemic’; seen in CT  Abundant ground substance, few cells and
 ELICITED fibers (Hallare, n.d.)
o Recruited from other areas; macrophages  Primary component of Wharton’s jelly of
going to area of inflamed, infected, or injured umbilical cord
CT b. ADIPOSE
 ACTIVATED  White Adipose for structural fill while highly
o Resident or elicited stimulated by foreign metabolic
material or infection to change morphology  Brown Adipose for heat regulation
and properties for increased activity c. RETICULAR
Mononuclear Phagocyte System (MPS) is the family of  Few collagenous and elastic fibers
monocyte-derived phagocytic cells distributed in various
 Support for motile cells and hematopoietic
organs.
tissues (lymphatic and myeloid)
d. ELASTIC
 For flexible support
e. PIGMENT found in some LCT)
f. LAMINA PROPRIA
 Its general characteristic is presence of many
lymphocytes

B. Connective Tissue with Special Properties

o ELASTIC TISSUE
 For flexible support
o HEMATOPOIETIC (lymphatic and myeloid)
 Includes macrophages, neutrophil, eosinophils,
lymphocytes, plasma cells and mast cells
o MUCOUS TISSUE
 ground substance rich in hyaluronic acid
 big portion is made up of macrophages and
Figure 4. Genealogy of blood cells
fibroblast, while collagenous and reticular fibers
make up a very small part of it
III. CLASSIFICATION OF CONNECTIVE TISSUE  major component of umbilical cord and found in
A. Connective Tissue Proper pulp of developing teeth
o LOOSE CONNECTIVE TISSUE
C. Supporting Connective Tissues
 Most widespread of connective tissues
 Loose randomly organized network of fibers in
o CARTILAGE
ground substance
o BONE
 Usually found in areas where little resistance to
stress is required; Occupies spaces around and
between muscles, underlies mesothelium IV. LABORATORY SLIDES
 Supports major internal organs and where 1. Ordinary Loose CT
blood vessels are distributed
o DENSE CONNECTIVE TISSUE
 Tightly packed fibers
 Little space occupied by cells and ground
substance
- REGULAR
 Fibers are parallel with one another
 Resist mechanical stress, (e.g tendon)
 Made up of parallel type I collagen fibers
- IRREGULAR
 Collagen fiber bundles are randomly
oriented
 Primarily collagenous fibers
 Found in dermis of skin, capsules of
spleen, liver and lymph nodes, sheath of
the large nerves
o MODIFIED LOOSE CONNECTIVE TISSUE
 Fibers are far apart but tissues have varying
histologic characteristics making it distinct from
ordinary areolar tissue. Figure 5. Ordinary Loose Connective Tissue

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2. Modified Loose CT - Lamina Propria
- Mucous CT  Slide: core of an intestinal villus
 Slide: umbilical cord

Figure 10. Lamina Propia found in the Intestinal Villus

Figure 6. Mucous Connective Tissue in the Umbilical Cord
3. Dense CT
- Reticular CT
- Regular Dense CT
 Slide: lymph node
 Slide: tendon

Figure 11. Regular Dense Connective Tissue in the Tendon

Figure 7. Reticular Connective Tissue in the Lymph Node - Irregular Dense CT

 Slide: pig skin
- Adipose Tissue

Figure 12. Irregular Dense Connective Tissue in the Pig Skin

Figure 8. Adipose Tissue

COMPONENTS OF CONNECTIVE TISSUE PROPER
- Pigment Tissue
1. Intercellular Fibers
 Collagenous Fiber
 Slide: Tendon Sheath

Figure 13. Collagenous Fiber in the Tendon Sheath

Figure 9. Pigment Tissue

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  Elastic Fiber  Macrophage

Figure 14. Elastic Fiber

Figure 18. Macrophage
2. Cells
 Fibroblast  Plasma Cell
 Slide: Protean cell

Figure 19. Plasma Cell

Figure 15. Fibroblast in the Protean Cell
 Mast Cell
 Adipocyte
 Slide: Omentum

Figure 20. Mast Cell

Figure 16. Adipocyte in Omentum  Neutrophil

 Mesenchymal cell

Figure 21. Neutrophil

Figure 17. Mesenchymal Cell

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  Lymphocyte  Pigment Cell

Figure 22. Lymphocyte

Figure 26. Pigment Cell
 Monocyte

V. References
Alvarez, MD, L. (2018). Connective tissue. Powerpoint presentation.
Britannica, T. E. (2017, December 13). Mast cell. Retrieved from
https://www.britannica.com/science/mast-cell
Lodish, H. (1970, January 01). Collagen: The Fibrous Proteins of the Matrix.
Retrieved from https://www.ncbi.nlm.nih.gov/books/NBK21582/
Marieb, E. & Hoehn, K. (2013). Human anatomy and physiology (9th ed.). USA:
Pearson Education, Inc.
Mescher, A. (2013). Junqueira's basic histology text and atlas (13th ed.). USA:
McGraw-Hill Education.
Wheeless' Textbook of Orthopaedics. (n.d.). Retrieved from
http://www.wheelessonline.com/ortho/chondroitin_and_keratin_sulfat
e

Figure 23. Monocyte

 Eosinophil

Figure 24. Eosinophil

 Basophil

Figure 25. Basophil

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