Neurobiology 104                                 Read G&H chaps. 4,6.

September 25,28 2003                              including pp 81-83

 

                        CONNECTIVE TISSUES

 

Connective tissues (C.T.) are the supporting tissues of the body. They:

    1. physically support the body: e.g. bone and cartilage

       and literally hold the body together.

    2. furnish pathways for nerves and blood vessels, and a

       compartment for exchange between capillaries and active cells.

    3. provide for immune functions, phagocytosis and wound healing.

C.T. comes in a great variety of forms, some highly specialized,

     e.g. dentin, cementum, bone, tendon, cartilage, fat, blood.

 

              I.  Embryonic: mesenchyme

             II.  Adult

                     A. Connective tissues proper ("ordinary")

                              Loose

                              Dense

                                  Irregular

                                  Regular

                     B. Supporting

                              Cartilage

                              Bone

                     C. Special

                              Adipose

                              Reticular - Lymphatic

                              Blood

                              Hemopoietic

 

All connective tissues have important common properties:

    Their cells are imbedded individually in extracellular matrix.

    They arise from embryonic ­mesenchyme­.

    They are interconvertible to some extent. 

         Even specialized types have intermediate forms.

    Connective tissues are categorized mainly by the physical

    properties of their extracellular components and not cells.

 

Ordinary Connective tissues

 

­Ordinary connective tissues­ are classified as ­loose­ vs ­dense­,

      and ­regular­ vs ­irregular­ mainly on basis of their fibers.

Looser C.T. has fewer, smaller fibers; more cells; more capillaries

    and more water.

Important examples of ordinary C.T. include:

    ­Lamina­ ­propria­ - The layer of C.T. underlying an epithelium.

    Dermis of skin - leather is tanned dermis.

    ­Stroma­ of organs - the framework tissue which supports the

    Parenchyma­ or specific active cells of organ.

        The physical texture and durability of an organ depend

        primarily on its stroma.

    Adventitia of organs and ­fascia­ between organs.

    Tendons and ligaments (and the cornea) are regular dense C.T.

       Note: Fibers can have various degrees of alignment in C.T.

       but only VERY organized C.T. is called "regular".

_______

Essential terms are bold


The 3 main structural components of ordinary C.T. are:

    collagen fibers, elastic fibers and amorphous ground substance.

    These are synthesized by ­fibroblasts­ in ordinary C.T.

 

Collagen

 

­Collagen­ is a unique fibrous protein:

    - the most abundant protein of the body (and of teeth).

    - very strong:  Tendons have a tensile strength of 20,000 lbs/sq in.

    - inelastic:  Wavy fibers can be straightened but not stretched.

    - evolutionarily ancient.

    - distinctive in chemistry and structure.

        Every third amino acid is glycine. 

        ­Hydroxyproline­ and ­hydroxylysine­ are almost unique to collagen.

 

Fibroblasts synthesize and secrete collagen as a procollagen precursor.

    Peptidases remove the ends extracellularly to form ­tropocollagen­.

 

 

See G&H Fig. 4-7      

 

 

 

    Tropocollagen is a helix with three intertwisted polypeptide chains.

       (2 alpha-1 and 1 alpha-2) specified by two separate genes

 

 

                           See G&H Fig. 4-5      

 

 

Tropocollagen chains aggregate in a staggered array to form

    a microfibril with cross bands 640 Angstroms (64 nm) apart,

    (the hallmark of collagen in the EM).

Microfibrils aggregate into collagen fibers (~1 micron across)

Collagen fibers associate into bundles of various thickness,

     Proteoglycans bind to the surfaces of fibrils, fibers and bundles

     to control their degree of polymerization.

Fibroblasts can deposit collagen in perfectly parallel arrays.

     They polymerize the fibers in grooves (“coves”) in their membrane­.

     They also stretch masses of collagen fibers to further align them.

 

At least 20 distinct types of collagen occur in different places:

     ­Type I­ is in ordinary C.T., tendons, bone and dentin.

     ­Type II­ occurs in hyaline cartilage.

     ­Type III­ forms very delicate fibers called ­reticular­ ­fibers­.

          Reticular fibers are very fine with enough carbohydrate to

             stain distinctively with ­PAS­ or silver ions.

          C.T. supported by reticular fibers is called ­reticular­ ­tissue­.

Type IV­ is found in basement membranes.  It does not form fibers.


 

­Elastica

­ 

  -  is a complex rubbery material of a cross linked globular protein.

  -  can have any shape; e.g. fibers, sheets and branched strands.

  -  can be specially stained because it is hydrophobic.

  -  complements the function of collagen by returning stretched tissue

        to its original state.

  -  seriously degenerates with age.

 

­                     Amorphous ground substance­ (read G&H pp 71-74)

 

is composed of gigantic proteoglycan molecules, with charged linear

    polysaccharides (­glycosaminoglycans­ = ­GAG­'s) attached to proteins.

Ground substance turns tissue fluids into extremely viscous fluids

    and gels because the ionized polysaccharide chains are extended.

       ­Hyaluronic acid­  mw ~l,000,000   produces soft gels.

       ­Chondroitin sulfate­  mw ~30,000  produces hard gels (cartilage).

       ­Keratan­ sulfate and dermatin sulfate are less abundant forms.

The functions of amorphous ground substance are:

    to convert the water in tissues into a gel.

    to immobilize particles but allow diffusion of small molecules.

    to regulate the amount and composition of the aqueous compartment.

    to attach cells and collagen fibers via adhesive glycoproteins.

       Adhesive glycoproteins attach cells to the extracellular matrix.

       Laminins anchor cells to basal laminae.

       Fibronectin holds cells to collagen and GAG’s.

 

 

                                  See G&H Fig. 4-3                 


II: THE CELLS OF CONNECTIVE TISSUE

 

Connective tissues contain a variety of cells that may develop locally or come in from the blood. They may be fixed in place or free to wander.

 

­Fibroblasts­ are the principal synthetic cells of C.T.

   They have a spindle shape, prominent nucleus and poorly visible

      cytoplasm.

   Active fibroblasts in loose C.T. have abundant cytoplasm,

      large pale nuclei, and retain the potential to divide and move.

   Inactive "fibrocytes" of dense connective tissue appear as dark

      elongate nuclei scattered between dense bands of fibers and

      probably no longer secrete, move or divide.

   Other cell types take over the synthetic functions of fibroblasts in

      specialized C.T.: smooth muscle, osteoblasts, reticular cells.

   Some fibroblasts partially develop the contractile properties of

      smooth muscle, so called myofibroblasts or myoid cells.

   Adult connective tissue encodes fixed tissue patterns.

   Fibroblasts look like their precursor mesenchymal cells but are

      an entirely different and more differentiated cell type.

   The best guess is that fibroblasts can differentiate into other

      C.T. cell types (eg adipocytes) but normally do not.

 

­Mesenchyme­ or embryonic C.T.

   Mesenchymal cells look like fibroblasts but have a great range of       developmental potentials.

   Most fetal mesenchymal cells have differentiated into restricted

      cell types in an adult but scattered ones remain, mostly along

      capillaries and are called pericytes. Pericytes can divide and

      become fibroblasts, adipocytes, endothelial cells, etc.

   Mesenchyme comes from mesoderm in the posterior body and from

      neurocrest in the head.

   C.T. cells arise from two sources:

      1. local mesenchyme – eg fibroblasts, adipocytes.

      2. via blood from bone marrow, white blood cells, mast cells

      Possibly some cells come from both sources (i.e. osteocytes)

 

­Macrophage­ (called ­histiocytes­ in ordinary C.T.):

   - ingest material by ­phagocytosis­ and pinocytosis, digest it with

        lysosomes and often retain indigestibles in residual bodies.

        They also have secretory functions and roles in immunity.

   - can be nearly as abundant as fibroblasts in loose C.T.

   - occur especially where needed the most

   eg under wet epithelia, in the spleen and in lymph nodes.

 

Histological appearances of macrophages.

   Macrophages comprise a variety of forms which vary in

      appearance (and, to an unknown degree, in function).

   They have special names in various tissues.

   The appearance of a macrophage depends upon mainly what it has

      ingested.

   New resting macrophages are hard to distinguish from fibroblasts

      but can be "vitally stained" by injecting dye into an animal.

   Old histiocytes often contain droplets of lipids ("foam cells").

   Macrophages are recognizable in electron micrographs by their

      abundant lysozomes and ruffled cell surface.


 

The life cycle of macrophages

 

   Precursors originate in bone marrow as ­monocytes­.

   Monocytes travel in blood, then migrate into C.T. and

differentiate into macrophages.

   Resting macrophages are fixed in place but become activated

        and migrate by chemotaxis during inflammation.

   Senescent macrophages can remain in C.T. for decades.

   Large indigestible particles cause macrophages to fuse into

           multinucleated ­foreign body giant cells­. 

 

             The ­Mononuclear Phagocyte System­MPS­)

 

Macrophages are best defined as the derivatives of monocytes.

These MPS cells also share certain biochemical commonalties.

   (e.g. cell surface receptors for antibody molecules and

    lysosomal enzymes.)

 

The cells of the MPS system include:

 

Histiocytes (in C.T.)

Free and fixed macrophages (lymph tissue, bone marrow)

Kupffer cells (liver)

Pleural and peritoneal macrophages (serous cavities)

Osteoclasts (bone)

Microglia (nervous tissue)

Langerhans cells (skin)

Foam cells (after inflammation)

Giant foreign body cells (inflammation)

 

            The "Reticuloendothelial System"

 

This is an outdated previous scheme that should be ABANDONED.

The "RE system" would include all phagocytic cells (except

    neutrophils) together with reticular and endothelial cells.

It was based on the mistaken interpretation that phagocytes

    differentiate from reticular cells and endothelial cells as

    well as from monocytes.

Unfortunately, you will hear doc's and even histologists speak of

   "RE cells".  Let's not have any dentists be so ignorant.

 

 

­Mast cells­:

 

  - cause inflammation, allergic symptoms and asthma.

  - are packed with secretory granules which contain

       ­histamine­, a vasodilator, and ­heparin­, an anticoagulant.

  - accumulate antibody molecules (IgE) on their surfaces as

    receptors.  They release their granules when these antibodies

    contact their antigens, to cause familiar allergic symptoms.

  - also secrete a variety of chemicals with roles in

    hypersensitivity reactions (skim through G&H pp 117-120).

  - share similarities with basophils of blood (and are sometimes

    called "tissue basophils") but are a different cell type.

  - tend to occur along blood capillaries.


­Adipocytes­ ("unilocular fat cells")

 

  - are large cells with a fat droplet and a narrow rim of cytoplasm.

  - function to store energy, insulate, produce heat and fill space.

  - occur as isolated cells, clumps, and large masses (adipose tissue).

  - develop from specialized preadipocytes or lipoblasts which resemble

    fibroblasts.

        Preadipocytes develop early in life and accumulate fat when

        you are old enough not to want it.

  - are surrounded by a basal lamina and supported by reticular fibers.

  - are metabolically active and complexly regulated.

        Adipose tissue is highly vascular.

    Human fetuses and infants have special brown fat.

        Its "multilocular" fat cells have many small fat droplets and

        mitochondria to generate body heat.

 

­Endothelial cells:

 

  ­ - line the entire blood vascular and lymphatic systems.

   - are very squamous with flat nuclei.

   - have great regenerative capacity.

   - rest on basal laminae which provide support and stability.

 

Capillaries fundamentally are components of C.T.

       They are confined to stroma (except in C.N.S.)

 

Endothelial cells are diverse to an unknown degree.

   They do much more than just passively line blood vessels.

   In some organs they have distinctive appearance and

      specialized defensive and secretory functions.

         Some have special cell surface proteins to draw

            leukocytes into the surrounding C.T. from blood.

         Some secrete chemical messengers to cause the smooth

            muscle in the media of arteries to constrict.

 

 

Leukocytes = White blood cells

 

These cells are transient invaders into C.T.

 

-->ALL BLOOD LEUCOCYTES DO THEIR WORK IN C.T. AND NOT IN BLOOD.<--

 

     ­Neutrophils­, ­Eosinophils­ and ­basophils­ infiltrate C.T. for

defense functions.

     Neutrophils are the main phagocytes for bacteria.

             The others have complex functions in immunity.

     ­Monocytes­ settle down as macrophages in C.T.

     ­Lymphocytes­ wander in large numbers in C.T. of vulnerable

     places.

        B-lymphocytes can give rise to ­plasma cells­ in C.T.

           Plasma cells are factories for making antibodies

           Plasma cells have much cytoplasm with basophilic rough ER.

        T-lymphocytes regulate other lymphocytes in various ways.

 

Wait for a later lecture on blood cells.

 

 


 

 

 

 

KODACHROME SLIDES SHOWN DURING LECTURE 1

 

 

       ­Slide­                             ­Purpose­

 

Mucosa of lip              contrast epithelium with C.T.

Mucosa                     vascularity of lamina propria

Skin                       organs embedded in C.T

Loose C.T.                 appearance at medium power

Dense C.T., high power     contrast with loose D.T.

Tendon:                    dense regular C.T.

EM of aligned collagen     show how regularly collagen can be

Area of C.T.               show spectrum of loose to dense

Nerve bundles H+E          show staining with H+E

Nerve, Masson stain        describe Masson stain

Cornea, Masson stain       dense regular C.T. with Masson stain

Large artery, H+E          distinguish smooth muscle and C.T. with H+E

Artery wall elastic stain  visualization of elastica

Liver, H+E                 appearance with H+E

Liver, Masson              visibility of collagen with Masson stain

Liver, silver stain        appearance of reticular fibers

Parotid gland, 0 power     show stroma

Submax. gland, med power   show stroma

Submax. gland, high power  show stroma

 


 KODACHROME SLIDES SHOWN DURING LECTURE 2

 

    Fibroblast in loose C.T.

    Diagram of active and inactive fibroblasts

    Fibrocytes in dense C.T.

    Mesenchyme from atlas

    Umbilical cord mesenchyme

    Diagram of a pericyte

    Diagram of the dual origin of C.T. cells

    Macrophage in bone marrow smear

    Macrophage in lymph node

    Macrophage in spleen

    macrophage in lung

    Loose C.T. with fibroblasts and possible macrophages

    Macrophage in C.T. spread identified by vital staining

    Medium power c.t. spread showing macrophage + fibroblasts

    Monocyte

    Table of the Monocyte Phagocyte System

    The reticuloendothelial system

    Tweedlee Dee's comment of the R.E. system

    Mast cells in C.T. spread

    Mast cells degranulated

    Fat cells

    Adipose tissue

    Brown fat

    Neutrophils in blood

    Neutrophils in C.T.

    Lymphocytes in blood

    Lymphocytes in C.T.

    Lymphocytes en masse in appendix

    Plasma cells

    Diagram of capillary

    Capillaries in fat tissue