Epithelium

Epithelium

Connective Tissue ridge (peg)

papilla

Connective Tisue

Muscle

CONNECTIVE TISSUE
To Discuss:
• General Structure and Function of Connective Tissue
• Embryonic Connective Tissue
• Ground Substance and ECM
• Connective Tissue Fibers
• Cells of Connective Tissue
• Connective tissue proper
– Types of Connective Tissues
 Connective Tissue
- provide a matrix that supports and physically connects
other tissues and cells together in organs. The interstitial
fluid of connective tissue gives metabolic support to
cells as the medium for diffusion of nutrients and waste
products.
- It is one of the most abundant and widely distributed
tissues in the body.
 General Structure
• One of the four basic types of tissues (epithelium, connective tissue,
muscle, and nervous tissue)
• Found throughout the body; most abundant and widely distributed in
primary tissues
• Connective tissues have:
– Mesenchyme Varying degrees of vascularity
– Nonliving extracellular matrix, consisting of ground substance and fibers
– cells (fibroblasts and others),
– fibers and ground substance (extracellular matrix)

Note: The amount of each of these components defines what type of

connective tissue is.
 Structural Elements
• All connective tissues share similar structural elements
• Composition:
Ground substance – unstructured material that fills the space
between cells
– Fibers – collagen, elastic, or reticular
– Cells – fibroblasts, chondroblasts, osteoblasts, and
hematopoietic stem cells
 Functions of Connective Tissue
• Protection by binding together and provide mechanical
support as architectural framework of the body (bone &
cartilage) and for other tissue (metabolic, defense,
transport, storage)
• Insulation (adipose tissue)
• Transportation
• Wound repair / inflammatory response
Connective tissue serves a variety of functions throughout the body.

Bacterium Fat cell

Macrophage
Connective tissue resists tension and compression

Collagen Elastin

Glycosamino- glycans
 Classification of Connective Tissue
1. Embryonic connective tissue
2. Connective tissue proper
3. Specialized connective tissue
 Embryonic Connective Tissue
• Embryonic messenchyme a gel-like ground substance with
fibers and star-shaped mesenchymal cells which gives rise
to the various connective tissue of the body.
– Found in the embryo
• Mesoderm- embryonic middle germ layer that particularly
gives rise to all connective tissue in the body.
Connective Tissue: Embryonic
 Ground Substance
• Gel-like, viscous, clear and slippery; high water content
• Part of the ECM that occupies space between cells and fibers.
Participates in binding cells to fibers.
• Consists predominantly of 3 groups of molecules:
– Proteoglycans, GAGs and multiadhesive glycoproteins
(fibronectin and laminin) 
• Interstitial (tissue) fluid functions as a molecular sieve through
which nutrients diffuse between blood capillaries and cells

Nice to know: it is lost in normal tissue preparation (extracted

during fixation and dehydration)
Ground Substance: Proteoglycan Structure
1. Glycosaminoglycans (GAG)
• linear (unbranched) polysaccharides, e.g. heparan sulfate, chondroitin
sulfate, keratan sulfate, hyaluronic acid
• very hydrophilic due to abundant negative charges (e.g. SO 4- groups).
• except for hyaluronic acid, are usually bound covalently to protein core as
part of a proteoglycan

2. Proteoglycans
• core protein + GAG side chains (like a bottle brush)
• bind cells, other proteins, and/or ECM components

3. Multiadhesive glycoproteins
• small glycosylated proteins containing NUMEROUS binding sites to cells,
signaling molecules, and other ECM components
• e.g. fibronectin and laminin: important for adhesion of epithelial cells to
the basal lamina via transmembrane integrin receptors.
 Ground
Substance
 Connective Tissue Fibers
• Collagen Fibers
• Elastic Fibers
• Reticular Fibers
 COLLAGEN
– is the most abundant class of proteins and pound for
pound some are as strong as steel (up to 30% dry weight)

• Collagens are a large family of proteins that form fibers or

networks.
• tough; provides high tensile strength

– multiple types:
a. fibril-forming or fibril-associated (in skin, tendon,
cartilage, bone, dentin, blood vessels);
b. cross-linked networks (in all basement membranes)
Fibrillar: type I, type II, type III

Network: type IV
 Major Collagen Fiber Types (out of at least 28)
Collagen Type Tissues Function

Fibril-forming collagens (these are visible)

I
Skin, tendon, bone, dentin Resistance to tension
(most abundant)

II Cartilage, vitreous of eye Resistance to pressure

III
Skin, muscle, blood vessels, liver, etc. Structural framework and stability
(reticulin)

Network-forming collagens

IV All basement membranes Support and filtration

Fibril-associated collagens with interrupted triple helices (FACIT)

VI, IX Assoc. w/ type I and II fibrils Fibril-fibril / fibril-ECM binding

Anchoring filament collagens

VII Epithelia Epidermis to basal lamina

Three collagens polypeptides associate to form rope-like
structures.
• Each of these polypeptides can be over 1000 amino
acids giving the trimer an overall length of 300 nm and a
width of 1.5 nm.
• The polypeptides wrap around each other and form
coiled coil interactions.
• The sequence is a repeat of 3 amino acids: glycine and
usually proline and lysine.
• Glycine which is the smallest amino acid allows for tight
packing of the polypeptides.
Fibroblasts synthesize and process collagen.
• Collagen is synthesize on ER-bound ribosomes and crosses the
ER membrane during translation similar to other secreted
proteins.
• In the ER two important modifications take place.
a. First, certain prolines and lysines are hydroxylated. These
modifications will allow for assembly into trimers and covalent
crosslinks between collagen trimers outside the cell.
b. Second, disulfide bonds between collagen polypeptides
mediates their assembly into trimers by facilitating interaction
between correct collagen proteins. One other feature of
intracellular collagen is that it contains extra sequence at its N
and C-termini called prodomains. These prevent collagen
trimers from assembling into fibrils inside the cell which would
be catastrophic for the cell.
Removal of prodomains allows collagens to assemble into
fibrils.
Lysyl oxidase crosslinks collagen trimers in fibrils.
 Collagen fibrils aggregate to form fibers.

All collagens are trimers but fibrous collagens assemble into parallel arrays to increase
mechanical strength. Fibrils then aggregate into large bundles called fibers -> type I.

This gives type I collagen three levels of interactions:

Trimer -> assembles inside fibroblasts.
Fibrils -> aggregation of trimers and crosslinking; outside fibroblasts.
Fibers -> aggregation of fibrils.
Collagen fibers viewed by light microscopy

H&E Trichrome
 Collagen fibers
viewed by
TEM*
Longitudinal

Transverse

*TEM, transmission electron

microscopy
 Clinical disorders resulting from defects in
collagen synthesis
Type Disease Symptoms
I Osteogenesis Spontaneous fractures, progressive
imperfecta hearing loss, cardiac insufficiency

III Ehlers-Danlos Hypermobility of digits, early

(type IV) morbidity/mortality from rupture of
aorta or intestine

multiple Scurvy (lack of vit. C, a Ulceration of gums, hemorrhages

cofactor for prolyl and
lysyl hydroxylase)
Ehlers-Danlos Syndromes
• A series of genetic diseases
with faulty assembly of
collagens (lysyl hydroxylase
deficiency).
• Hyperextensible skin and
hypermobile joints
• In some forms (e.g., type IV),
weakness in blood vessels
or intestines are life
threatening.
Type collagen Disease
I Osteogenesis imperfecta
II Kniest dysplasia
III Ehlers-Danlos type IV
IV Alport’s syndrome
VII Kindler’s syndrome
IX Multiple epiphyseal dysplasia (MED)
X Schmid metaphyseal chondrodysplasia

XI Stickler’s syndrome type II

XVII Generalized atrophic benign epidermolysis
bullosa (GABE B)
ELASTIC FIBERS
 Elastic Fibers
LM: Visualized by selectively staining with Weigert’s, resorcin-fuchsin, or
aldehyde-fuchsin

EM: Consist of amorphous core of elastin surrounded by microfibrillar

glycoprotein, fibrillin (8-10nm).

Elastin: is rich in glycine and proline, but it contains little or no hydroxyproline

and hydroxylysine . uniquely contains desmosine and isodesmosine, which
are thought to cross-link the molecules into a network of randomly coiled
chains. This cross-linking is responsible for its rubber-like properties.

Confers elasticity: present in large amounts in ligaments, lung, skin, bladder,

and walls of blood vessels.

Marfan Syndrome: defect in elastic fiber synthesis; reduced elasticity in skin

and lungs, skeletal defects (bones are longer and thinner than usual),
cardiovascular complications (aneurism, valve prolapse)
Elastic fibers are a composite of elastin and fibrillin
• Elastic fibers are a composite material composed of two
primary components: elastin and fibrillin.
• Both are synthesized by fibroblasts and secreted into the
surrounding tissue where they assemble into elastic fibers.
• Fibrillin fibers are thin and arranged in more or less parallel
arrays. They are required for correct assembly of elastic
fibers.
• Elastin appears as a amorphous substance. Elastin is what
gives elastic fibers its characteristic mechanical properties
of stretching and recoiling.
Network of elastin molecules can stretch and
recoil like a rubber band
Reticular (Reticulin) Fibers

• Form a delicate supporting framework for

highly cellular tissues (endocrine glands,
lymph nodes, liver, bone marrow, spleen,
smooth muscle).
• Composed mainly of Type III collagen,
with a carbohydrate moiety that reduces
Ag+ to metallic sliver = argyrophilic.
• Special stain: silver impregnation to
visualize.
• Thinner than type I collagen (Type III
fibrils are 30-40 nm diameter; type I fibrils
are ~200 nm diameter)
 Reticular Fibers (type III collagen)
• made by reticular cells (specialized fibroblasts) and vascular smooth muscle cells
 Cells in Connective Tissue
• Each Type of connective tissue has to be made by a certain
cell type. In an immature stage each cell below secrets the
fibers needed for its connective tissue.
• Fibroblasts – connective tissue proper
• Chondroblasts – cartilage
• Osteoblasts – bone
• Hematopoietic stem cells – blood
• White blood cells, plasma cells, macrophages, and mast
cells
 Resident

Pollard et al. Cell Biology 2nd Edition Transient

 Cells in Connective Tissue
1. Fibroblasts
Fixed 2. Adipose (fat) cells
(permanent
residents) 3. Tissue Macrophages**
4. Mast cells**

5. Lymphocytes & Plasma Cells

Free (differentiated B-cells) **
(transient
residents) 6. “Leukocytes”**
(specifically, neutrophils, eosinophils, & basophils)
** derived from hematopoietic stem cells and
involved in immune function and inflammation
Connective Cell Lineages
Transient cells digest connective tissue to aid motility and migration.

• Because many cells in connective tissue come from blood,

they must cross several barriers and migrate through
connective tissue. First they must cross the endothelial
cells. They must also cross the basement membrane and
migrate through the connective tissue. Most immune cells
will digest components of connective tissue to open gaps
through which cell can move. Metastatic cells develop a
similar ability to escape a tumor and find their way to
lymphatic or blood vessel.
Matrix metalloproteinases are a family of proteins that digest components
of connective tissue.

• Cells that navigate across basement membrane and

through connective tissue utilize set of proteases to digest
components called matrix metalloproteases (MMPs).
MMPs require zinc as a cofactor and can be membrane-
bound or secreted outside cell as soluble proteins. There
are different types of MMPs that digest different types of
collagen in connective tissue. Cells also secrete elastase
which digests elastin.
Fibroblasts are the most common cells in connective
tissue
• Synthesize and secrete components of the ECM: fibers
and ground substance.
• Active and quiescent stages (when quiescent sometimes
called fibrocytes or mature fibroblasts).
• Synthesize growth factors.
• Rarely undergo cell division unless tissue is injured,
which activates the quiescent cells.
• Play a major role in the process of wound healing and
respond to an injury by proliferating and enhanced fiber
formation.
Active and inactive fibroblasts
Adipocytes predominate in adipose tissue
• Very active cells with many functions:
• Triglyceride storage and glucose metabolism (insulin
and glucagon receptors)
• Secretion of many bioactive molecules:
 leptin (regulates satiety)
 angiotensinogen (blood pressure)
 steroids (glucocorticoids & sex hormones)
 growth factors (e.g. insulin-like growth factor, tumor necrosis
factor )
 cytokines (e.g. interleukin-6)
Single, large
lipid droplet

White (common, yellow, unilocular) adipose tissue stained with Masson’s trichrome
 Adipocytes

Lipid (fat)
droplet

Nucleus

Capillaries
 Brown (Multilocular) Adipose Tissue
• Large mesenchymally derived cell specialized for cytoplasmic
storage of lipids.
• The cells are polygonal and generally smaller than white
adipose tissue. Their nuclei are often centrally located.
• The brown colour is mainly due to very abundant mitochondria
(containing cytochrome) and the large no of capillaries.
• The many lipid droplets, mitochondria and capillaries all help
mediate the principal function of brown adipose tissue, which
is thermogenesis in non shivering mammals such as newborns,
who have a substantial amount of brown adipose tissue.
 Brown (Multilocular) Adipose Tissue

• Present in newborns (and hibernating

mammals) and involved in thermoregulation
• Mitochondria of brown fat cells express uncoupling
protein which “short circuits” the electron transport
chain producing HEAT rather than ATP.
 Uniocular/white adipocyte
• Large mesenchymally derived cell specialized for cytoplasmic
storage of lipids. Termed uniocular because the triglycerides are
contained within a large single lipid droplet which takes up the
majority of the cell.
• Because of its large size, the lipid droplet displaces and flattens the
nucleus against the cell membrane, giving it a signet ring
appearance with a thin rim of cytoplasm also associated with the
nucleus.
• Most of the cytoplasm surrounds the nucleus and contains small
golgi, RER etc. The membranous layer surrounding the lipid
droplet contains cisternae of SER and pinocytotic vesicles.
white

brown
 Macrophage
• Structure:
– they are part of the “mononuclear phagocyte system“(MPS/RES)
– 30 micrometer, ruffled membrane (irregular shapes), acidophilic
lysosomes in the cytoplasm; can have various heterogeneous
“inclusions” ingested material
– Round, oval or kidney-shaped, eccentric nucleus; can have
nucleoli
– Derived from the peripheral blood monocytes, involved in
phagocytosis and inflammatory response
– Has various shapes, localizations and names
• Function:
– Mainly phagocytosis
– Triggered by a specific interaction between membrane receptors
and ligands
– Consequences:
a. cell movement towards target particle
b. pseudopodia formation – engulfment
c. respiratory burst
d. secretion: cytokines, interferon, completement and
e. coagulation factors
f. production of matrix metalloproteinases
Monocytes escape from blood vessels into connective tissue where they
differentiate into Macrophages

Primary function: phagocytosis and antigen presentation

 PLASMA CELLS
• Structure:
– Ovoid/pear-shaped cells
– Basophilic cytoplasm (due to abundant RER), with a perinuclear pale area
(Golgi apparatus); can contain acidophilic Russel bodies (secretory granules)
– Eccentric nucleus, with hetero and euchromatin in a characteristic pattern:
“cart wheel” or “clockface” (the heterochromatin resembling the spokes of
the wheel or the numbers on a clock), visible nucleolus.

Function:
– Found in lymphoid organs (lymph nodes, spleen, bone marrow) and
connective tissues associated to the respiratory and digestive mucosae 
– Originate in B lymphocytes, that are terminally differentiated as a response to
antigen challenge 
• Secrete immunoglobulins (antibodies): IgM, IgG, IgA, IgE,IgD
 Plasma Cells are mature B lymphocytes
that constitutively secrete antibodies

Black arrows indicate several plasma cells White arrows = Golgi regions
 Mast Cells
• Principal function is storage in
secretory granules and REGULATED
release (degranulation) of histamine
and other vasoactive mediators of
inflammation.
• Responsible for the immediate
hypersensitivity response
characteristic of allergies, asthma
and anaphylactic shock.
• Connective tissue mast cells are
found in skin (dermis) and peritoneal
cavity; mucosal mast cells are in the
Metachromasia – when stained with toluidine blue, the mucosa of the digestive and
granules bind the dye and change its color to red. respiratory tracts.
 Important molecules released by mast cell:
• Heparin
• Histamine
• Serine proteases
• Eosinophil and neutrophil chemotactic factor
• Cytokines
• phospholipids
 Neutrophils
• Enter connective tissue from blood vessels as the “first
wave” in acute inflammatory responses
• Small cells with multi-lobed, heterochromatic nuclei (aka
“polymorphonuclear neutrophils”, “PMNs”, “polys”)
• Primary function: anti-bacterial (are phagocytic like
mphages, but SHORT-lived and NOT antigen presenting)
 Connective tissue proper
• Besides bone, cartilage and blood all mature connective
tissues belong to the Connective Tissue Proper class.
• We can break these into loose connective or dense
connective.
 The density and organization of fibers determines the strength of
connective tissue.

Dense Regular Dense Irregular Loose

 Connective Tissue Proper: Loose
• Areolar connective tissue
– Gel-like matrix with all three connective tissue fibers
– Fibroblasts, macrophages, mast cells, and some white blood
cells
– Wraps and cushions organs
– Widely distributed throughout the body
 Connective Tissue Proper: Loose
• Adipose connective tissue
– Matrix similar to areolar connective tissue with closely packed
adipocytes
– Reserves food stores, insulates against heat loss, and supports
and protects
– Found under skin, around kidneys, within abdomen, and in
breasts
– Local fat deposits serve nutrient needs of highly active organs
 Connective Tissue Proper: Loose
• Reticular connective tissue
– Loose ground substance with reticular fibers
– Reticular cells lie in a fiber network
– Forms a soft internal skeleton, or stroma, that supports other
cell types
– Found in lymph nodes, bone marrow, and the spleen
 Connective Tissue Proper: Dense Regular
• Parallel collagen fibers with a few elastic fibers
• Major cell type is fibroblasts
• Attaches muscles to bone or to other muscles, and bone to
bone
• Found in tendons, ligaments, and aponeuroses
Connective Tissue Proper: Dense Regular
 Connective Tissue Proper: Dense Irregular
• Irregularly arranged collagen fibers with some elastic fibers
• Major cell type is fibroblasts
• Withstands tension in many directions providing structural
strength
• Found in the dermis, submucosa of the digestive tract, and
fibrous organ capsules
 Connective Tissue: Cartilage
• Hyaline cartilage
– Amorphous, firm matrix with imperceptible network of collagen
fibers
– Chondrocytes lie in lacunae
– Supports, reinforces, cushions, and resists compression
– Forms the costal cartilage
– Found in embryonic skeleton, the end of long bones, nose,
trachea, and larynx
 Connective Tissue: Elastic Cartilage
• Similar to hyaline cartilage but with more elastic fibers
• Maintains shape and structure while allowing flexibility
• Supports external ear (pinna) and the epiglottis
Connective Tissue: Fibrocartilage Cartilage
• Matrix similar to hyaline cartilage but less firm with thick
collagen fibers
• Provides tensile strength and absorbs compression shock
• Found in intervertebral discs, the pubic symphysis, and in
discs of the knee joint
 Connective Tissue: Bone (Osseous Tissue)
• Hard, calcified matrix with collagen fibers found in bone
• Osteocytes are found in lacunae and are well vascularized
• Supports, protects, and provides levers for muscular action
• Stores calcium, minerals, and fat
• Marrow inside bones is the site of hematopoiesis
 Connective Tissue: Blood
• Red and white cells in a fluid matrix (plasma)
• Contained within blood vessels
• Functions in the transport of respiratory gases, nutrients,
and wastes
 Classification of connective tissue
• Embryonic Connective Tissue
-Messenchyme
-Mucous Connective Tissue
• Connective Tissue Proper
-Loose Connective Tissue
-Dense Connective Tissue
Regular
Irregular
• Specialized connective Tissue
-Adipose Tissue
-Blood
-Bone
-Cartilage
-Hematopoietic tissue
-Lymphatic tissue
 LIBRARY WORK
• IN A YELLOW PAPER:
• List down (and familiarize yourself for future reference):
a. Cells of the mononuclear phagocytotic system
b. Most common collagenopathies in human
-Collagen type
-Disease
-Symptom
Deadline: Friday 1pm sharp
THANK YOU!