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Histology lecture #7

Connective tissue #2

Done By Yazeed Al-Ajlouni

Last lecture we started talking about the connective tissue and in this lecture we will
finish this chapter.

So I mentioned there were 3 main types of fibers in the connective tissue which are:

1-collagen 2- reticular 3- elastic fibers

The first two are formed by a collagen molecule or protein whereas the 3rd one is
composed mainly of elastin, and I showed you last time this electron micrograph, but
just to see the 3 fibers simultaneously (at the same time) so that you can know the
differences between them.

So this is an electron micrograph of collagen fibrils, we said that there was a class of
collagen where the molecules aggregate forming collagen fibers.

When you look at collagen fibers in the EM you see the collagen fibrous that form the
fibrous because you know you can see the finest or the most detailed structures of
the cell in the EM, so here you have collagen fibrous (in slides photo).

These are longitudinally cut whereas these ones are transversely cut.

In the longitudinally cut section of collagen fibrous, you can see alternating light and
dark bands, whereas here it is the transversely cut collagen molecules and then you
can see the cut ends of the collagen molecules that formed the collagen fibrous at
that level.

This is a H&E stain section under the light microscope , you can say that collagen is
the only fiber that you can see in the H&E section and it's very eosinophilic, and you
can see those collagen bundles looking pink because of the eosin stain.

Of course the other structures here, the elongated structures are the fibroblast
which produced subunits for collagen fibers formation.

The 2nd type of fibers I told you about last time was reticular fibers, remember
reticular fibers are agrayophilic which means that they love silver; they cannot be

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seen using H&E stain. However, they are seen in sections which are stained with
silver using impregnation method).

Remember that reticular fibers appear as thin or fine threads, they form networks
that provide attachment sites for the cells, both of these here and here are just
silver-stained sections, showing those fine threads of reticular fibers that form the
network.

The 3rd type is the elastic fibers, reticular fibers are thinner than collagen and also are
the elastic fibers, the elastic fibers add resiliency to connective tissue by forming
sparse networks in organs that are subject to much bending or stretching.

They occur in fenestrated sheets forming elastic laminalae in large arteries such as
the aorta (laminalae means layers, laminalae is the plural form of laminala {layer}).

Elastic fibers are composed of 2 structures, the 1 st one is the elastin protein, the 2nd
one is fibrillin , fibrillin is a glycoprotein that aggregates forming microfibrils, so in the
completely formed elastic fibers you can see a deposit of elastin in the center with
fibrillin microfibrils at the surface, as you will see in the Electron micrograph.

You have to know that fibrillin microfibrils predominate over the elastin in
developing elastic fibers and the opposite is true in adult fibers were the elastin
predominate.

Elastin is secreted by fibroblast in the connective tissue and smooth muscle cells in
the walls of large arteries.

As I said before that procollagen matures in the extracellular matrix is the same case
for elastin, so elastin matures in the extracellular matrix.

Although elastin is resistant to digestion by most proteases, it's easily hydrolyzed by

pancreatic elastase, elastase it lyses (it breaks down elastin).

These are Electron micrographs illustrating the process of elastic fibers formation.

Here it's just like those fibrillin micro fibrils that are forming the elementary or the
developing of fibrillin fibers before the deposition of elastin.

Now in the 2nd one you can see those deposits, the amorphous deposits of elastin
with the fibrillin microfibrils.

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 Here in the last stage the elastin accumulates in the center and then you have
fibrillin microfibrils at the surface.

In order to view or see elastic fibers under the light Microscope it has to be stained
or the sections have to be stained with the special stains, here in this image on the
left you can see those dark purple colored fibers, those are the elastic fiber among
the collagen bundles, the collagen bundles are still eosinophilic so it's special stain in
addition to H&E.

In the image on the right you can see those fenestrated sheets or lamillae of elastic
fibers, so those things here the brownish structures are the elastic fibers forming
lamillae.

You can see other structures here, elongated grayish in color these are smooth
muscle cells .because these are lamellae, they are in the wall of the large arteries so
the cells producing those elastic fibers subunits are the smooth muscle cells, you will
take that later, don’t be concerned whether these smooth muscle or fibroblast,{ they
are just the cells secreting the subunits for elastic fibers} .

The 3rd component of the extracellular matrix of the connective tissue is ground
substance, the ground substance is highly hydrated transparent complex mixture of
macromolecules which are principally glycolsaminoglycans refer to as GAGs ,
proteoglycans ,and multi adhesive glycoproteins.

Due to the viscosity of the ground substance it acts as a lubricant and barrier to the
penetration of invaders, this is an Electron Microghraph of part of connective tissue
you can see this is the border of the cytoplasm of fibroblast, this is the nucleus and
this is the cytoplasm with the braff cells. here those circular structure are the
collagen fibrils forming the fibers and this is the elastin protein forming the elastic
fibers.

In between intervening like the cells the fibers both collagen and elastic fibers you
have this granular material filling the spaces ,now this represents the ground
substance, the ground substance can be relieved in the TEM as either spaces filled
With granular substance or empty spaces.

Glycoseaminoglycans are sulfated polysaccharided composed by repeating

disaccharidea units.

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examples of glycoseaminoglycans are :

-dermatan sulfate , condraytan sulfate ,keratan sulfate –heparan sulfate .

Hyaluronic acid is one of the glycoseaminoglycans that is considered the largest and
most ubiquitous, it's very unique because it's the only one that it's not sulfated. all
the other glycoseaminoglycans are sulfated.

Hyaluronic acid is synthesized in extracellular matrix by the hyaluronate synthase

( synthase end with ase but it is not for the lysis, not for the break down because the
prefix perceiving ase is not a substrate for break down ,synthase for synthesis ) it's
for hyaluronic acid synthesis.

Proteoglycans are macromolecules composed of a core protein with sulfated

glycoseaminoglycans attached to it. you can say that a Proteoglycan is composed of
core protein and GAGs which are polysaccharides, so what is the different between
Proteoglycans and glycoproteins? (remember when I talked about membrane there
were the glycoprotein which were the peripheral proteins and carbohydrates with
polysaccharide attached to them ), So the difference between the Proteoglycan and
glycoprotein is the proportion of polysaccharide and proteins or which predominates,
so in Proteoglycan there are more carbohydrates than in the glycoprotein, where as
in the glycoprotein the proteins predominate where as the polysaccharide are just
additional.

so Proteoglycan similar to the glycoprotein are synthesized in the rough ER, they
mature in Golgi apparatus and they are secreted by cells by exocytosis, now one of
the Proteoglycan that you need to know is the agricane which is the dominant
Proteoglycan present in the cartilages.

Proteoglycan serves as structural component, they anchor cells to the matrix and
they bind unsequestive surface signaling proteins which are important, for example
for growth and differentiation of fibroblast.

Multi adhesive glycoproteins are macromolecules where the protein moiety be

predominate so they are still glycoprotein,

They function in the adhesion of cells to their substrate, ex of the multi adhesive
glycoproteins include:-

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1-Laminine, you need to know this because it's abundant in the basal lamina, it
adheres the epithelial cells to the lamina

2- fibronectin which is synthesized by fibroblasts and some epithelial cells.

There are the matrix receptors which are the cells surface molecule that mediate the
interaction of cells to the extracellular matrix by binding to: collagen, fibronectin, and
laminine, they are called integins, these matrix receptors are the integrins and you
know that the integrins are Transmembrane linker proteins.

Types of the connective tissue:

- The embryonic and the adult connective tissue.

The embryonic is the mesenchyme and the mucus, the mesenchyme which has
mesenchymal cells which are the undifferentiated cells that give rise to all types of
connective tissue as well as other cell types.

Adult connective tissue can be loose, dense, or specialized; loose is also called areolar
connective tissue.

The difference between loose and dense is the proportion of the components, so in
loose connective tissue you have equal parts of the 3 components cells, fibers,
ground substance, whereas in dense connective tissue you have more collagen fibers,
you can see the bundles very conspicuously.

The dense connective tissue can be either irregular or regular based on the
orientation of the collagen fiber bundles, so if the collagen fiber bundles are in
certain orientation, parallel to each other then it is regular.

Whenever they are in each direction, with no definite orientation it's irregular.

Specialized could be reticular or adipose (adipose= fat cells from adipocytes.)

Mucous tissue is an embryonic tissue, it's a jelly like tissue, it's rich in ground
substance with much hyaluronic acid,

It has scattered fibroblast with few collagen fibers.

You can see mucous tissue in the umbilical cord where it's refer to as wartans jelly,
you can also see it in fetal tissues in the embryonic tissue, and you can see it in the
pulp cavity in young teeth.

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The 1st type of the adult connective tissue is loose or areolar connective tissue, the
loose connective tissue supports epithelial tissue, so expect to see it adjacent to the
epithelial tissue, that’s why whenever you see epithelium the layer immediately
subjacent (underlying the epithelium) should be loose areolar connective tissue, it
also forms a layer around small blood and lymphatic vessels and it fills the space
between muscle and nerve fiber.

Fibroblast and macrophages are the most numerous cells but there are other cells
present as well, known that all types of fibers are present in the loose connective
tissue: reticular, elastic, and collagen fibers.

So the dense irregular connective tissue has much collagen fibers which predominate
over the ground substance, these collagen fibers are arranged in bundles in all
direction without definite orientation and therefore they provide resistance to a
stress which can come from all the directions.

the dense irregular connective tissue is often closely associated with loose
connective tissue where one can grade into the other, so for ex in the skin you can
see the epithelium, underlying it is loose connective tissue and then the dense
connective tissue.(as you can see in the slides images)

Ex of a dense connective tissue are tendons and ligaments which hold the
components of the muscular skeletal system, tendons attach muscle to bones, and
ligaments attach bones to each other.

This is an Electron micrograph of one of the fibrocytes. How you can tell this is
fibrocyte? Look there's very sparse cytoplasm and you have those process, you can
see this nucleus here and you can see it somewhat heterochromatic, because you
know that the fibrocytes are the quiescent fibroblast so they are the inactive stage,
so when they are inactive they do not need to synthesize anything, that’s why they
do not have much rough ER or well-developed Golgi apparatus, even the nucleus is
not euchromatic, because if the nucleus is euchromatic this indicate high synthetic
activity of the cell.

Reticular tissue consists of reticular fibers, and you know that the reticular fibers are
formed by type III collagen and they are produced by specialized fibroblast called
reticular cells.

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Reticular tissue is present in hematopoietic and lymphoid organs, we well learn it
later.

Reticular tissue contains cells of mononuclear phagocyte system such as

macrophages.

This is a schematic diagram or drawing of the reticular tissue, you can see those fine
threads, the reticular fibers forming network and providing framework for the cell
attachment.

Here of course in this network the blood circulate.

Here is a silver stain section, and you can see those dark looking or black-colored
threads forming the network and then there are the cells attaching at this network.

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Done by: yazeed al-ajlouni

Forgive me if there is any mistake, I couldn't get the slides.