EMBRYONIC CONNECTIVE TISSUE

CONNECTIVE TISSUE PROPER

ADIPOSE TISSUE
 Categories of connective tissues
q Embryonic connective tissues
ØMesenchymal
ØMucous

q Adult connective tissues q Specialized connective tissues

ØConnective tissue proper ØSupporting tissues
§ Loose (alveolar) § Cartilage
§ Reticular § Bone
§ Dense irregular § Blood
§ Dense regular
ü Collagenous
ü Elastic
§ Adipose
 CONNECTIVE TISSUES
Ø They have the same origin – mesenchyme and the same structure
(cells and extracellular matrix),
Ø Unlike epithelial cells, connective tissue cells are widely seperated
by components of extracellular matrix.

Epithelial
tissue
Closely aggregated polyhedral cells
and very little extracellular matrix
Connective Epithelia are derived from all germinal
tissue layers

Cells of connective tissue + extracellular matrix (ECM)

 Connective tissue is composed of two elements:
ØCells
ØExtracelullar matrix [ECM]
ü Fibers -collagen fibers, elastic fibers, reticular fibers
ü Ground substance

Extracellular matrix = Fibers + ground substance

Cells of connective tissues (CTs)

v Connective tissue proper - fibroblasts, fibrocytes
v Adipose tissue – adipoblasts, adipocytes
v Cartilage – chondroblasts, chondrocytes
v Bone – osteoblasts, osteocytes
v Blood – formed elements (erytrocytes, leukocytes)
 Cells of Connective Tissue (CT) proper

ØThe cells spend all their live in the tissue

§ Fibroblasts – originate from undifferentiated mesenchymal cells,

produce ECM components, can proliferate

§ Fibrocytes – quiescent fibroblasts

Two stages of activity

The active cells The quiescent cells

¯ ¯
Fibroblasts Fibrocytes
ü Fibroblasts Fibroblasts _ Fibrocytes
§ the most common cells
in connective tissue ü Fibrocytes
§ branched cytoplasm § smaller than
§ ovoid, large and pale fibroblasts
staining nucleus with § fewer processes
nucleolus § smaller, darker,
§ rich in RER and well elongated nucleus
developed Golgi § small amount of
complex RER
§ responsible for the
synthesis of ECM
components
§produce the growth
factors
§ proliferate when the
additional fibroblasts are
required
 The function of connective tissues

Ø Providing structural support

Ø Serving as a medium for exchange
Ø Aiding in the defense and protection of the body
Ø Forming a site for storage of fat

The function of connective tissue proper

Ø Forms the capsules of organs and membranes that enveloped the

central nervous system (meninges)
Ø Forms trabeculae and walls inside several organs – components
of the stroma within organs
Ø Medium for exchange of metabolic waste, nutrients and oxygen
between the blood and many of cells in the body
The cells which reside in the tissue

§ Macrophages
§ Mast cells
§ Plasma cells
§ Leukocytes
f 10 – 30 mm Macrophages ® the Mononuclear Phagocyte System
ØMorphological features reflect functional activity of macrophages:
§ they have pseudopodia, only in macrophages
§ irregular surface with and protrusions – pinocytotic and
phagocytic activities
§ oval or kidney-shaped nucleus located centrally
§ well-developed Golgi complex, many lysosomes, prominent
rough endoplasmic reticulum (RER)

ü Macrophages derive from monocytes of blood

ØMacrophages are long-living cells, can proliferate locally. The cells are distributed
throughout the body, and in certain region have special names:
§ histocytes – the connective tissue proper
§ Kupffer cells – the liver
§ microglia cells – the central nervous system
§ osteoclasts – the bone
 Macrophages functions
Ø phagocytosis of foreign substances and bacteria
Ø antigen processing and presentation to other cells
(Antigen Presenting Cells – APC)
Ø secretion of cytokines and other molecules
§ enzymes, eg. collagenase
Ø removing cell debris and damaged extracellular components

Ø Macrophages when stimulated

§ may increase in size and are arrangement in clusters forming epithelioid
cells;
§ may fuse to form multinuclear giant cells
Ø Epithelioid cells and giant cells are found only in pathological conditions.

Multinuclear giant cells

 Mast cells
derive from cells of the bone marrow
f 10 – 13 mm

Ø Oval to round connective tissue cells

Ø The cytoplasm is filled with basophilic secretory granules
Ø Small, spherical nucleus, situated centrally

v Secretory granules of mast cells

§ they contain biological active substances:
ü histamine - promotes an increase in vascular permeability
ü heparin - (sulfated glycosaminoglycan is blood anticoagulant)
ü neutral proteases
ü chemotactic factor for eosinophils

§ substances not stored in the granules:

ü leukotriens (C4, D4, E4)
 Mast cells
q The connective tissue mast cells
§ skin and peritoneal cavity

q The mucosal mast cells

§ the connective tissue of the intestinal
mucosa and lungs
vThe surface of mast cells contains specific receptors for immunoglobulin E
(IgE), a type of immunoglobulin produced by plasma cells
 Leukocytes
or white blood cells
Øthe wandering cells of connective
tissue
Øthey migrate through the walls of
capillaries and postcapillary
venules from the blood to
connective tissue (diapedesis) Diapedesis
v Granulocytes
v B and T Lymphocytes
ØPlasma cells Adipose cells
Ø Adipose cells, adipocytes, fat cells
§ the connective tissue cells for
storage of neutral fats or for
Ø B lymphocyte ® plasma cell ® synthesis the production of heat
of immunoglobulins
Ø large, ovoid cells with basophilic cytoplasm,
very well developed RER, The Golgi complex
Ø spherical nucleus, placed eccentrically
Ø average of live is short, 10 – 20 days.
 The extracellular matrix

ØFibers ØGround substance

§ Collagen fibers § Glycosaminoglycans (GAGs)
§ Elastic fibers § Proteoglicans
§ Reticular fibers § Multiadhesive glycoproteins

Fibers

v Collagen fibers
ü are formed by protein collagen
ü the collagen is the most abundance protein in the human body (30% of dry weight)
ü the collagens belong to a family of more than 25 members, produced by several
cells type
üare acidophilic; stain pink with eosin, blue with Mallory’s stain

eosin Mallory’s
 ØThe cell type responsible for collagen synthesis
§ fibroblasts, chondrocytes, osteoblasts
§ odontoblasts
§ endothelial cells
§ vascular smooth muscle cells
ØThe collagen profile
• the principal amino acids – glycine (33.5%), proline (12%), hydroxyproline (10%)
• amino acids that are characteristic of the collagen – hydroxyproline and hydroxylysine

Collagen types
v Type I - the abundant: skin, tendon, bone, dentin
v Type II - cartilage, vitreous body
v Type III - skin, muscle, blood vessels
v Type IV - all basement membranes
v Type V - fetal tissue, skin, bone, placenta
v Type VII – epithelia – anchors skin epidermal basal lamina to underlying stroma
(anchoring fibrils)
Collagen is a protein polymer composed of monomeric units of the protein
Tropocollagen
Tropocollagen Three a chains

§ elongated protein 280 nm in length and 1.5 nm in width

Structural arrangement of collagen:

1) Collagen is arranged into microfibrils.
2) Microfibrils are arranged into fibrils.
3) Fibrils are grouped into a fiber.
4) Fibers are grouped into a collagen bundle
 Collagen synthesis
v Procollagen synthesis
§ Registration peptides on both
amino-terminal and carboxy-
terminal ends
§ precursor of tropocollagen

v Hydroxylation of proline and lysine

§ peptidyl proline hydroxylase
§ peptidyl lysine hydroxylase
Co-factors: O2, Fe, vit. C

v Glycosylation of hydroxylysine
§ transferases
Mn

v Removing of registration peptides

§ procollagen peptidases
§ tropocollagen

v Formation of covalent cross-links

between tropocollagen molecules
§ lysyl oxidase
Cu and O2 ions
Vitamin C (ascorbic acid) deficiency leads to the scurvy, disease characterized
by the degeneration of connective tissue. Scurvy characterized by bledding
gums and loose teeth among other symptoms
Vitamin C deficiency ® abnormal hydroxylation of procollagen ® synthesis of
defective collagen
 Reticular fibers

Øconsist mainly of type III collagen

Øextremely thin
Østain black by impregnation with silver salts – agyrophylic fibers

ØLocalization
§ are particularly abundant in smooth muscle tissue
§ endoneurium
§ spleen, lymph nodes, bone marrow
§ constitute a network around the cells of parenchymal organ, eg. liver,
endocrine glands
§ the wall of arteries

Reticular fibers
 Elastic fibers
Ø Elastin – the main component + microfibrills
Ø Proelastin - precursor of elastin
Ø Elastin is rich in glycin and proline
Ø Contains two unusual amino acids – desmosine and
isodesmosine formed between four lysines
Ø has rubberlike qualities
Ø stains brown with orcein and violet with
resorcine-fuchsin

They are composed of an amorphous elastin

surrounded by a microfibrillar component
consisting of fibrillin

v Elastoblasts
§ Fibroblasts
§ Chondrocytes
§ Vascular SMC
§ Endothelial cells
 The elastic fibers system

ØOxytalan fibers ØElaunin fibers ØElastic fibers

§ zonule fibers of the eye
§ dermis
q do not contain elastin
q consist of a bundle of 10nm
microfibrils composed of
glycoproteins:
fibromodulin I and II,
and fibrillin.
§ around sweet glands
§ dermis
q irregular deposits of
elastin between
the microfibrils
§ the wall of large arteries
§ connective tissues
q elastin located
centrally and thin
sheath of microfibrils
q the most numerous
component of
the system

Microfibrils

Elastin
 Ground substance
Øhighly hydrated, colorless and transparent complex mixture of macromolecules
Øit fills the space between cells and fibers
Øit is viscous and acts as lubricant and barrier to the penetration of invaders

v Glycosaminoglycans (GAGs)
v Proteoglycans
v Multiadhesive glycoproteins
 Glycosaminoglycans (GAGs)

Øcalled mucopolysaccharides

- dermatan sulfate
- chondroitin sulfate
- keratan sulfate
- heparan sulfate
- hyaluronic acid

Øwith the exception of hyaluronic acid GAGs are bound covalently to a protein
core, forming proteoglycans
Øwith the exception of hyaluronic acid all other GAGs are sulfated
ØGAGs are intensely hydrophilic and act as polyanions
 Proteoglycans
Ø are composed of a core protein associated with the four main GAGs (without
hyaluronic acid)
Ø proteoglycan (monomer) is three-dimensional structure (can be pictured as test
tube brush)
q Proteoglycans of extracellular matrix (ECM)
§ aggrecan – the most important, the
dominant in cartilage
§ syndecan, fibroglycan

Proteoglycan Functions:
monomer ü structural component of ECM
ü anchoring cells to the ECM
Core protein ü as extracellular and surface
proteoglycans bind many protein
growth factors (TGF-b
® transforming growth factor)
Proteoglycan aggregates
v Proteoglycans are degradated by several
cell types (lysosomal enzymes).
Lysosomal enzymes deficiency causes
several disorders in humans
Schematic representation
(eg. Mucopolysaccharidoses )
of the proteoglycan aggrecan
 Multiadhesive glycoproteins

Ø play an important role in the interaction between neighboring

adult and embryonic cells
Ø play role in the adhesion of cells to their substrate

v Fibronectin
§ the product of fibroblasts and epithelial cells
Protein
§ has sites to bind cells, collagen and GAGs

v Laminin
§ participates in the adhesion of epithelial cells to basal lamina

v Matrix receptors are cell-surface molecules. Cells bind to collagen, fibronectin,

laminin
§ Integrins – transmembrane linker proteins, interact with the cytoskeleton
v Intracellular proteins – the interaction between integrins, ECM, cytoskeleton
elements
§ Paxilin
§ Vinculin
§ Talin
 Embryonic connective tissues
Mesenchyme

Ø The precursor embryonic tissue for all types of connective tissue

Ø Stellate undifferentiated cells and ground substance
Ø The lack of fibers
Ø Under specific stimuli the cells differentiate into the cells of connective tissue -
fibroblasts, adipoblasts, chondroblasts, osteoblasts, blood cells
 Embryonic connective tissues
Mucous
Ø Mucous connective tissue
§ has a abundance of ground substance rich in hyaluronic acid
§ is jellylike tissue containing very few collagen fibers type I and type III
§ the cells – mainly fibroblasts
§ is found in umbilical cord and is referred to as Wharton’s jelly
§ is found in the pulp of young teeth

Umbilical cord
 Adult connective tissues
The connective tissue proper
Loose connective tissue

ØLoose connective tissue

§ the very common type of connective tissue
§ fills spaces between groups of muscle cells, supports epithelial tissue, forms
layer around lymphatic and blood vessels.
§ is found in dermis, hypodermis, mucous membranes
§ has delicate consistency, well vascularized, not very resistant to
stress
Ø Reticular connective tissue
§ consists of reticular fibers (type III collagen) and specialized fibroblasts named
reticular cells
§ reticular tissue creates the special microenvironment for hematopoietic organs
and lymphoid organs (bone marrow, lymph nodes, spleen), and envelops
certain cells (eg. hepatocytes)

Reticular cell

Reticular
fibers
 Dense connective tissue (CT)
Øis adapted to offer resistance and protection
Øthere are fewer cells than loose connective tissue and high amount of collagen fibers

ØDense irregular CT
§ the collagen fibers are arranged in bundles without a definite orientation
§ provide resistance to stress from all directions
§ is found in dermis

Collagen fibers Fibrocytes

ØDense regular CT (collagenous)
§ the collagen bundles are arranged in the definite pattern
§ it is found in tendons

Tendon
§ the collagen fibers have parallel, closely packed bundles of
collagen separated by a small quantity of intracellular
ground substance
§ fibrocytes have elongated nuclei parallel to the fibers
§ the cytoplasm of fibroblasts stains the same color as the
fibers

Nuclei of fibrocytes

Collagen fibers
Ø Dense regular CT (elastic)
§ is composed of bundles of thick parallel elastic fibers
§ the spaces between the fibers are occupied by thin collagen fibers and flattened
fibroblasts
§ is found in yellow ligaments of the vertebral column, suspensory ligament of the penis

Aorta

fibrocytes

Elastic laminae beetwen them

vascular smooth muscle cells
Adipose tissue
 ØSpecial type of the connective tissue with predomination of adipose cells
(adipocytes, fat cells)
ü Adipocytes
§ as isolated cells, in small aggregates (adipose tissue)

Adipose tissue is found in a variety of places Functions:

such as: 1] stores energy
1] the hypodermis 2] contributes to the thermal
2] surrounding and protecting certain organs insulation of the body
3] the medullary cavity of long bones 3] protects delicate organs from
mechanical trauma
Adipose tissue

v Unilocular adipose tissue v Multilocular adipose tissue

(common or yelow) (brown)
 Unilocular (common or yelow) adipose tissue

ØThe color varies from white to dark yelow (depends on diet – carotenoids)
ØThe distribution is determined by age and sex
§ in the newborn has uniform thickness in the body
§ its distribution is partly regulated by sex hormones and adrenocortical hormones
(different distribution in male and female body)

Spherical, isolated cell

Signet ring cells Polyhedral
cells in
adipose tissue
Lipid droplet Sudan III

Lipids are removed

in routine
Nucleus histological
techniques
Cytoplasm
 Unilocular (common or yelow) adipose tissue

ØCytoplasm
f 50 – 150 mm § Golgi complex
§ mitochondria
§ poorly developed RER
§ SER
§ free polyribosomes
§ pinocytotic vesicles

Basal lamina

Vimentin intermediate filament

v Adipose tissue
ü is richly vascularized
 Unilocular (common or yelow) adipose tissue

ØLarge depot of energy for organism

Ø Metabolism of adipose tissue is regulated by hormones

Adipose tissue as a secretory organ

Ø enzymes – lipoprotein lipase
Ø leptin
Ø adiponectin
Ø adipsin
ØTNFa (Tumor Necrosis Factor a)
Ø source of estrogen (estrone) in postmenopausal women
Multilocular adipose tissue (brown fat)
ØMultilocular tissue cells are polygonal and smaller than
unilocular adipose cells
ØThe cytoplasm contains
§ great number of lipid droplets of various sizes
§ spherical and central nucleus
§ numerous mitochondria with abundant long cristae
(containing colored cytochromes)
ØThe cells receive direct sympathetic innervation
ØThe tissue is rich in capillaries
 Distribution of multilocular adipose tissue

Ø In humans the tissue is important mainly in the first postnatal life (produces
heat and protects the newborn against cold)
ØIt is greatly reduced in adulthood
ØThe tissue is more abundant in hibernating animals – hibernating gland

In human newborn the multilocular adipose tissue constitutes 2 - 5% of the body weight

v Mainly around the shoulder blades and kidneys

 Function of the multilocular adipose tissue

ØThe main function is to produce heat

ØThe production is stimulated when human newborn or animals are exposed to
the cold environment
ØMitochondria of multilocular adipocytes contain transmembrane protein –
thermogenin ® heat production (temperature of tissue is elevated)
 Slides

v Mesenchyme

Embryo head
ØMucous tissue

The umbilical cord - Warthon’s jelly

Vein

Arteries
 ü Nuclei of fibrocytes and others cells
ØLoose connective tissue ü Collagen fibers
ü Elastic fibers

Collagen fibers

Elastic fibers
ØDense connective tissue

v Dense regular CT v Dense irregular CT

Tendon Skin
ØAdipose tissue

Sudan III

Signet ring cells