Connective Tissue

Okay,we will start with connective tissue. I mentioned things about connective tissue when I
talked about the stroma,right? I said that it is usually the connective tissue. The connective tissue
supports and binds other tissue and it is composed of cells and extracellular matrix. The extracellular
matrix is composed of protein fibres and ground substance. The protein fibres can be
collagen,reticular and elastic. The connective tissue develops from an embryonic multipotential
mesenchymal cells. The mesenchyme is formed by cells called mesenchymal cells which are
elongated undifferentiated cells. The mesenchymal cells give rise to all types of connective tissue as
well as other types of structures like the endothelial cells, blood cells and muscle cells.Of course,
remember that the mesenchyme arises from the middle layer of embryo which is called mesoderm.
Do you know the 3 layers of the embryo:ectoderm, mesoderm and endoderm?

Ok, remember that the embryo is composed of 3 layers, ectoderm (ecto is outside so its the
external), mesoderm(it’s the middle layer), and than the endoderm(it’s the innermost layer). So the
mesenchyme arises from the middle layer which is the mesoderm.

The connective tissue contains variety of cells with different origins and functions. There are too
many groups rising from 2 different origins. The first one arises from the mesenchymal cells which
are present in the mesenchyme which is an embryonic tissue. The other one arises from the
hematopoietic stem cells of the bone marrow.

So this diagram represent the cells that arises from the mesenchymal cells. You have first the
fibroblast, the adipocytes or fat cells, chondroplast, chondrocytes and osteoplast and
osteocytes.This cells here smooth muscle cells and endothelial cells and mesophilial cells arise from
undifferentiated mesenchymal cells but they are not connective tissue cells.

So,the other groups arises from the precursor cells of the bone marrow called the stem cells. You
have the plasma cells, the macrophage, langerhan cells, microphilia and osteoblast which are
macrophage-like cells and you have the mast cells and leucocytes. Of course the red blood cells and
megakaryocytes(you know that megakaryocytes is produced in the platelets), both of these are not
connective tissue cells.

The fibroblast are those connective tissue cells which are involve in the synthesis of the component
of extracellular matrix. They can exist in two forms. The first one is the active,the second one is the
quiescent. Quiescent means inactive/inert. Now, the active fibroblast have large, ovoid eukromatic
nucleus and prominent nucleoli with abundant RER and well developed Golgi Apparatus in the
cytoplasm. All of these things are indicating the active synthetic activity of those fibroblast. So
compared to the active fibroblast you have the quiescent fibroblast with smaller, fewer processes
and the nuclei are smaller, darker and elongated.

So this is a stained section of the connective tissue.Its from the dermis. The quiescent fibroblast are
referred to as fibrocyte.Now, fibroblast and fibrocyte can be distinguished morphologically.So based
on their morphology, you can see for example these cell, it’s a spindle shaped, with large, oval
eukromatic nucleus and here is the prominent nucleolus. This is another one.

This are fibroblast. Notice that the cytoplasm in the fibroblast is somewhat basophilic due to the
abundant of RER. If you compare the cytoplasm of the fibroblast with fibrocytes, its more acidophilic
in the fibrocytes due to the presence of less RER. And then the nucleus of the fibrocytes is
elongated,darker and smaller. Adipocytes are also presents in the connective tissue and they are the
fat-storing cells and here in this section you can see adipocytes(fat cells). You can see how the lipid is
stored in the adipocytes as a single droplet compressing all the organelles and the nucleus to the
periphery of the cells.

Macrophages arises from hematopoietic cell of the bone marrow which give rise to monocytes. The
monocytes circulates in the blood vessels and then penetrate the ore crust (p/s:not sure with this
‘ore crust’) through the walls of the venules or capillaries to penetrate the connective tissue where
they transform into the macrophages by increasing the cells size, increasing protein synthesis and
the number of Golgi Apparatus and lysosomes. Macrophages are sometimes referred to as
histiocytes.They are members in the mononuclear phagocytes system. So they are long living cells
which are involved in the uptake processing and presenting antigens for lymphocytes activation.

There are macrophage-like cells in different organs such as copper cells in the liver, microglia in the
CNS , langerhan cells in the skin, osteoblast in bone tissue.

This is electron micrograph of the macrophages. First you can see the protrusion here and
endingtations at the surface of the macrophages indicating its pinocytotic and phagocytotic activity.
You can see many lysosomes which are these black spots here, spherical in shape. These are the
lysosomes. Then you can see this nucleus which are oval or kidney in shape. So this is the
macrophages. Of course, there are many Golgi complexes or apparatus but you can barely see any
because of the low magnification. However I can tell you about this one here.

Mast cells originate from hemapoeitic stem cells of the bone marrow which circulate in the blood
and then cross the wall of the blood vessels penetrating the connective tissue. Now, according to
their abundance, or the area where they are most abundant,there are 2 populations of
macrophages. The first one is perivascular and the second one is mucocells. Peri means around.

So this population is the one around the blood vessels.And then the mucocells so it is in the mucosa
in the organ. Mast cells are involved in the release of bioactive substances that mediate the innate
immunity, inflammatory respons and tissue repairs.

Examples of these bioactive substances are here.

 Heparin,which act as anticoagulant.

 Histamine which promotes increase vascular permeability and the smooth muscle
contraction.

 Serine proteases which activates vias mediator of inflammation.

 Eusinophils and the neutrophils chemotactic factors.When you say chemotactic so its
attracting.So these are the factors that attract eusinophils and neutrophils.

 Leukotriene ,C4,D4 and E4 which trigger smooth muscle contractions.

This is a stained preparations or sections showing mast cells. All of these are mast cells.

You can see the basophilic secretory granules. Mast cells have secretory granules which are strongly
basopihillic in their cytoplasm and these secretory granules are the one that contain the bioactive
substances which I mentioned in the previous slide. So you have round or spherical nucleus which
can be sometimes abscured by this granules-like in this cell. This an electron micrograph of the mast
cells. This is the plasma membrane of the mast cells.You can see the electron dense granules which
are heterogenous. They are heterogeneous because they contain different substances.

So whenever you see a cell in electron micrograph containing these granules,this should be the mast
cells.

You know that the plasma cells are the cells that produce immunoglobulins. They arise from B-
lymphocytes. So since they have synthetic activity, you expect them to have RER, abundant RER
therefore their cytoplasm is basophilic and they have well developed golgi apparatus. Thats why
they have this pale, just a nuclear area here so its occupied by the golgi apparatus as well as the
centrioles. The last characteristic of the plasma cells is its nucleus.It is essentrically located. When
you’re saying essentrically located, its means that its peripheral in its location. So its situated close
to one periphery of the cells. And here this an EM of the plasma cells. You can see the essentrically
located nucleus and here are RER, see how abundant the RER is. This is a unique appearance of
nucleus of the plasma cell. You can see alternating areas of dark heterokromatin with lighter
eukromatin giving a clock phase appearance.

Leucocytes are the WBC . They migrate from the blood vessels by diappetices. This process is called
diappetices. So diappetices is the outward migration of the WBC from the blood vessels and this
migration is greatly increased during inflammation. Leucocytes do not return to the blood except the
lymphocytes.

Now, connective tissues contain fibres and these fibres are composed by protein molecules which
aggregate to form this fibres. And there are 3 main types.

1.collagen

2.reticular

3.elastic

Both collagen and reticular fibres are formed by collagen proteins whereas the elastic fibres are
composed mainly of elastin protein. There are more than 20 types of collagens. They are classified
into 4 types:

1.collagen which form long fibrils

2.fibrils associated collagen

3.collagen which form anchoring fibrils

4.collagen which form networks

So regarding the first class which is the long fibrils forming collagen you have those collagen
molecules which assemble to form elongated structures called fibrils and example of long fibrils
forming collagen is collagen type 1. The second class which is the fibrils associated collagen exist like
short structure which binds the fibrils to each other as well as to other components in the
extracellular matrix.

In the 3rd class, you have the collagen molecules forming the anchoring fibrils which binds or anchor
the basal lamina to the reticular fibres in the underlying connective tissue. So this should be type 7
collagen. Do you remember in the basement membrane,where you have those 2 layers (the basal
lamina and reticular lamina)? you need the anchoring fibrils formed by type 7 collagen to anchor or
bind the epithelium basal lamina to the underlying connective tissue.

Now the last one is the collagen molecules which form network.And this is found in the basal lamina
of the epithelium.And an example of this is type 4 collagen.

This is an EM of collagen fibrils.You have 2 cut collagen. The 1st one is longitudinally cut fibrils and
the 2nd one is transversly cut. For the longitudinally cut fibrils, you can see alternating light and dark
area and from the transversely cut, you can see the end of one collagen molecule. This is a stained
section under LM. You can see the elongated cells here with nucleus and the cytoplasm, this are the
fibroblast and this are the collagen bundles formed by the collagen fibres which are formed by the
collagen fibrils, which are in turn formed by the collagen molecules. Now remember that collagen
fibres are eusinophillic and so they look pink.

Now, we are going to look into collagen synthesis (this is for collagen 1) or the long fibrils forming
synthesis.

Don’t pay attention to the details in the book, just an overall summary of it as I will mention. You will
be asked about this not about its synthesis in the book. So, the first information is provided by mRNA
for the synthesis of polypeptide chain required for synthesis of collagen 1 in the RER. Then, this
polypeptide chains form triple helix. The triple helix is called tropocollagen which moves to the Golgi
apparatus and eventually out of the cells forming the tropocollagen molecules which aggregates to
form the collagen fibrils. Of course collagen turnover is very common in periodontal ligaments which
are the ligaments surrounding the teeth or the root of the teeth. For collagen turnover, the collagen
have to be degraded by collagenase. Now ,maybe you know that whenever you see an enzyme
ending with ‘ase’, so its breaking things down, according to the first part of the enzyme or the word.

So collagenase is an enzyme that breaks down collagen and then after that there is further
degradation by nonspecific protease so that then, more new collagen molecules are synthesised.
Reticular fibres are formed mainly by type 3 collagen and they contain more sugars than do the
collagen fibres, thats why they are PAS-positive and agyrophillic. Agyrophillic means, love silver. So
they are stained by silver impregnation appearing as black or dark brown lines. The reticular fibres
are formed by fibroblast-like cells which are called reticular cells. These are 2 sections which are
stained by the silver impregnation. You can see those fine lines. These are the reticular fibres.The
dark spots here are just the nuclei. The same as here. These are threads, fine reticular fibres. Those
are just the nuclei.

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p/s:I’m really sorry for this not well done lecture writing because of absence of Histology slides which
I can refer to. While reading, you should really look at the slides because a lot of things that was
explained is referring to micrograph, diagram, pictures and they are all in the slides. Supposed that i
want to put the diagram/pictures/micrograph images from the slide according to explanation in this
lecture notes but the slides are not there. So, try your best by referring to the slides by yourself.
About the spelling of scientific terms, again, refer to the slides. Just to be sure. Thanks a lot.

Done By: Siti Fatima