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12/16/10

Debuque, Isulat
LABORATORY ACTIVITY
CONNECTIVE TISSUE PROPER

Draw your observations of the following CT types. Caption your drawings


with the appropriate tissue types, and source of specimen. Label all
structures seen (fixed and wandering cells, matrix components, blood
vessels, nerves, membrane coverings).

a. Loose (areolar) CT b. Dense regular white fibrous CT

c. Dense regular yellow elastic CT d. Dense irregular CT

e. Reticular CT f. Mucous CT

g. Embryonic CT h. Adipose tissue


STUDY-FOCUSING QUESTIONS:

1. Compare and contrast the following by tabulating the required information:


a. CT proper types and their: predominant cell types, predominant matrix
component, ground substance present, function, location

Loose (areolar) CT Dense regular CT Dense irregular CT

Predominant Varied: Sparse cellular component: Macrophages, mesenchymal


cell types Resident fibroblasts, immigrant Fibroblasts, tendon cells in cells.
macrophages, mast cells, tendons
formed elements of blood
(except RBC and platelets).

Predominant Watery extracellular matrix, Dense masses of fibers Dense random arrays of fibers.
matrix contains relatively few running in parallel/orthogonally
extracellular fibers. arranged layers.
component

Ground Amorphous, contains Amorphous, contains Amorphous, contains


substance proteoglycans and proteoglycans and proteoglycans and
glycoproteins glycoproteins glycoproteins
present

Function

Location Lamina propria, mesenteries Ligaments, tendons, stroma of Dermis of skin, capsules of
the cornea. organs e.g. liver and spleen,
periosteum surrounding bones.

b. CT fiber types and their: protein composition, arrangement of protein


subunits, arrangement and appearance of fibers in tissues, physical properties
and function, staining properties, distribution

Collagen Fibers Yellow or Elastic Fibers Argyrophyl or


Reticular Fibers

Protein 3 polypeptide chains held Amorphous elastin, elaunin and Identical to collagen.
composition together by hydrogen bonds oxytalan. Micorofibrils. (Precursors of type I
and III collagen.)

Arrangement Occur in thick bundles or as Elastin molecules exist as cross- Not branched. Not
of protein individual fibrils. linked random coils. When cross- wavy as the
linked, they form a rubber-like collagenous fibers
subunits
network of molecules that returns to when released from
its original state after distortion.The tension.
microfibrillar protein assembles into
anastomosing networks of fibrous
proteins surrounded by amorphous
elastin.

Arrangement Running in all directions in a Appears yellowish, highly refractile, Unlike collagen, they
and wavy course; dull and opaque in homogeneous, and are not made up are thinner and form
appearance. Fibers bundled of fibrillar subunits that are visible delicate networks
appearance
together branch and anastomose; with the light microscope. instead of thick bundles
of fibers in individual fibers do not branch.
tissues
Collagen Fibers Yellow or Elastic Fibers Argyrophyl or
Reticular Fibers

Physical Supports most tissues and gives Distend when stressed and then The fibers form a soft
properties cells structure from the outside. return to their original shape when skeleton (stroma) to
main component of fascia, the stress is released. Major support the lymphoid
and function
cartilage, ligaments, tendons, component of many blood vessel organs (lymph nodes,
bone and teeth. Along with soft walls. In the wall of the aorta, elastic red bone marrow, and
keratin, it is responsible for skin fibers form numerous concentric spleen). (The thymus is
strength and elasticity, and its fenestrated elastic laminae. Exist in the only lymphoid
degradation leads to wrinkles that elastic cartilage, the lungs and organ that does not
accompany aging. It strengthens pleural membranes, the vocal cords, contain reticular
blood vessels and plays a role in and elastic ligaments. connective tissue.)
tissue development. It is present Adipose tissue is held
in the cornea and lens of the eye together by reticular
in crystalline form. fibers.

Staining Easily stained by ordinary dyes. Stains poorly with standard ionic They react with silver
properties Exhibit acidophilic properties in H dyes since it contains a few charged stains and with periodic
& E stained sections. In sections amino acids; as such it requires the acid-Schiff reagent but
stained with Masson’s trichome, special Verhoeff’s (Weigert’s are not demonstrated
they stain green. WIht Mallory’s resorcin fuschin) stain or Halmi’s with ordinary
trichome stain, they stain blue. aldehyde-fuchsin method. histological stains such
Thin fibers (type III) stain darkly as those using
with silver stains, thicker bundles hematoxylin.
do not.

Distribution Most numerous in all connective Found in the skin, lungs, arteries, Abundant in regions
tissues. veins, connective tissue proper, around blood vessels,
elastic cartilage, periodontal muscle fibers, fat cells,
ligament, fetal tissue and other basement membrane of
structures. epithelia, endoneurium,
lymphoid organs and
red bone marrow.

c. Collagen types and their: distribution, site of synthesis, main function,


molecular organization (fibril-forming, etc.)
Distribution Site of Synthesis Main Function Molecular Organization

Type I Found in tendons, Intracellular. Fibril-forming. 3 polypeptide chains held


ligaments, bone, together by hydrogen
dermis, organ bonds
capsules, and loose
connective tissue.

Type II Found in embryonic Intracellular. Fibril-forming. 3 polypeptide chains held


notochord; in adults, together by hydrogen
only in the nucleus bonds
pulposus of the
intervertebral disk,
vitreous body of the
eye, and in the
hyaline and elastic
cartilage matrix.

Type III Abundant in loose CT, Intracellular. Fibril-forming. 3 polypeptide chains held
in soft and compliant together by hydrogen
organs such as the bonds
walls of blood
vessels, in the stroma
of spleen, kidney and
uterus, and compose
the reticular laminae
underlying epithelial
basal laminae.

Type IV Found in basal intracellular. Network-forming. 3 polypeptide chains held


laminae. together by hydrogen
bonds

Type V Present in external Intracellular. Fibril-forming. 3 polypeptide chains held


lamina of muscle together by hydrogen
fibers, epithelial basal bonds
lamina, blood vessels
and in small amounts
elsewhere.

Type VI Present where types I Intracellular. Unknown. 3 polypeptide chains held


and III are found; together by hydrogen
kidney, uterus, bonds
cornea.

Type VII Form anchoring fibrils Intracellular. Anchoring. 3 polypeptide chains held
of epithelial basal together by hydrogen
lamina, serving to bonds
stabilize and firmly
anchor the epithelium
to the dermis.

Type VIII Major component of Intracellular. Unknown. 3 polypeptide chains held


Descemet’s together by hydrogen
membrane, the bonds
corneal epithelium
basal lamina.

Type IX Found mainly in Intracellular. Fibril-associated. 3 polypeptide chains held


cartilage. together by hydrogen
bonds

Type X Found in the matrix Intracellular. Unknown. 3 polypeptide chains held


surrounding together by hydrogen
hypertrophic bonds
chondrocytes of
degenerating growth
plate cartilages in
sites of future bone
formation.

Type XI Found in cartilage. Intracellular. Fibril-forming. 3 polypeptide chains held


together by hydrogen
bonds
Type XII Found in tendon Intracellular. Fibril-associated. 3 polypeptide chains held
fibroblasts. together by hydrogen
bonds

d. CT cell types and: morphological characteristics, synthetic activity, mitotic


activity, main function

Morphological Synthetic activity Mitotic Main function


characteristics activity

Fibroblasts Varies from fusiform to Secretions include Have a single Secrete extracellular
stellate, depending on its collagen, fibronectin, nucleus with matrix, bind
packing arrangement glycoproteins, and several extracellular matrix
relative to extracellular proteoglycans. nucleoli. constituents to form
fibrils. Extracellular tissue, and facilitate
glycoconjugates. wound healing.

Macrophages Highly variable shape due Monokines, IL-1, Have a single Dedicated phagocytes
to movement throughout enzymes, nucleus with and are distributed
CT. Diameter of approx. complement proteins, several throughtout the body.
20 µm and a single regulatory factors. nucleoli. They are derived from
irregularly shaped nucleus bone marrow stem
with one or two prominent cells and can assume
indentations and a several different forms.
conspicuous mass of Phagocyte at sites of
euchromatin. infection.

Mast Cells Similar to basophils in Releases histamine, have a single Involved in


peripheral blood. slow reacting nucleus with inflammatory reactions
substance, and several and immediate
heparin. (Preformed nucleoli. hypersensitivity allergic
mediators, newly reactions.
formed lipid
mediators, and
cytokines.)

e. Cell types of Mononuclear phagocyte system and their: morphological


characteristics, contents of cytoplasmic granules (if any), major secretory
product (if any), role in immunity, location
Morphological Contents of Major Role in immunity Location
characteristics cytoplasmic secretory
granules (if any). product (if
any).

Bone marrow Small cell body with N/A N/A Capability of giving Bone
stem cells few cell processes that rise to indefinitely marrow
are long and thin.. more cells of the
Large round nucleus, same type and
prominent nucleolus. other cells by
differentiation.

Monocyte Large phagocytic Internal vesicles. Cytokine. Replenish Bone


white blood cell with macrophages and marrow/s
simple oval nucleus dendrites. pleen.
and clear, grayish
cytoplasm.

Stellate cells of Stellate cells. N/A Inflammatory Early ethanol- Liver


Kupffer cytokines, induced liver injury sinusoids
collagen. in chronic
alcoholics.

Alveolar/free 15-25 µm in diameter, Particulate carbon. Pulmonary Destory foreign Serous


macrophages euchromatic nucleus, surfactant. material. cavities
irregular surface that
bear filopodia.

Microglia May be amemboid or N/A Myelin. Constantly CNS


ramified. Changing scavenge the CNS
shapes with oblong for damaged
nuclei. neurons, plaques,
and infectious
agents.

Osteoclasts Large multinucleate Vesicles and Cathepsin K Absorbs bone Bone


bone cell. 40 µm in vacuoles. tissue during growth
diameter. and healing.

f. Specialized types of CT and their: morphological characteristics, predominant


cell types, distribution, function
Morphological Predominant cell types Distribution Function
characteristics

Reticular CT Stellate reticular Reticular cells, resident Stroma of liver, bone Form a soft
cells have long macrophages adhering marrow, spleen, skeleton to support
cytoplasmic to fibers, few plasma lymph nodes, thymus. the lymphoid
extensions that join cells, RBC and WBC. organs. Adipose
other cells. tissue is held
together by reticular
fibers.

Embryonic CT Very cellular. Highly Fibroblasts of the Fetal life, dental pulp, Support the
vascular. spindle and stellate tunica propria developing fetus.
variety. Undifferentiated
mesenchymal. Some
blood cells and
phagocytes.

Mucous CT Soft jelly-like Fibroblasts, few Embryo, especially Forms the umbilical
homogeneous macrophages, lymphoid under the skin. cord.
ground substance is wandering cells. Limited to the dermis
very abundant. and hypodermis in
Contains granules adult animals.
and fibrillar
precipitates.

Adipose Presence of Adipocytes. Widely distributed in Stores fat, provide


rounded clear the body as fat insulation against
spaces which are deposits. heat loss, and
closely spaced, mechanical support
separated by the in certain regions of
thin fibrous strands the body.
of both collagenous
and elastic fibers.

g. Unilocular and multilocular adipose tissue and their: basis for coloration, lipid
distribution, precursor cell type, predominant organelles, vascular supply,
innervation, distribution in the body, functional capabilities

Unilocular adipose tissue Multiocular adipose tissue

Basis for coloration Contains mostly of lipids. Rich vascularization and large quantity
of mitochondria.

Lipid distribution Approximately 60-85% in one big fat Distributed in several small fat vacuoles
vacuole. embedded in mitochondria and other
organelles.

Precursor cell type Fibroblast-like precursor cells Fibroblast-like precursor cells

Predominant organelles Single central fat vacuole. The nucleus is Rich vascularization and large quantity
flat and unremarkable. of mitochondria. Small fat vacuoles.

Vascular supply Minimal vascularization. Rich vascularization.

Innervation Minimal innervation. Richly innervated.

Distribution in the body Bulk of body fat. Prevalent over the Hibernating species and newborn
abdomen and around the hips and human infants.
buttocks.

Functional capabilities Heat insulation, mechanical cushion, Generates heat in infants.


source of energy.

2. Describe the role played by the following enzymes in CT fiber synthesis,


assembly, and turnover:

a. Collagenase - Break the peptide bonds in collagen

b. Elastase - Determines the mechanical properties of CT.


c. Signal peptidase - Allows for the signal peptide to be cut off.

d. Lysyl oxidase - Catalyzes formation of aldehydes from lysine residues in


collagen and elastin precursors, this results in cross-linking collagen and
elastin for stabilzation of collagen fibrils for integrity and elasticity of mature
elastin.

e. Procollagen peptidase - Involved in the intracellular processing of collagen


where procollagen are deposited in the cisternae.

f. Proline hydroxylase - Aids in the production of hydroxyproline, an essential


element of collagen, a necessary element of connective tissue.

3. List the effects of the following on CT structure and function:

a. hydrocortisone
1. Aids in fat, protein, and carbohydrate metabolism.
2. Stimulates gluconeogeneis, the breakdown of protein and fat.

b. ACTH
1. Increases the bio-availability of cholesterol in the cells of the
adrenal cortex.
2. Increases the transport of cholesterol in the mitochondria and
activates its hydrolysis.

c. hypothyroidism
1. Low activity of the thyroid gland causes retardation of growth, thus
low production of connective tissue.
2. The connective tissue gains less nutrients for it to develop properly
due to low level of metabolism.

d. Ascorbic acid
1. Ascorbic acid is involved in the hydroxylation of proline to
hydroxyproline, an important step Ascorbic acid is involved in the
hydroxylation of proline to hydroxyproline, an important step in the
synthesis of collagenin the synthesis of collagen.
2. Without ascorbic acid, underhydroxylated molecules are produced,
thus producing poor collagen fibers. Usually results in scurvy.

4. List the factors that lead to lipogenesis, lipolysis.

Lipogenesis: Acetyl-CoA is added with two carbon atoms in the mitonchodria of liver
cells.

Lipolysis: Triglycerides hydrolyze into three fatty acids followed by further


degradation in to acetyl units by beta oxidation.