St. Luke’s College of Medicine – William H.

Quasha Memorial
ANATOMY BLOCK 1

Lecture: 5- Intro to Histology Date: August 5, 2015

Lecturer: Catherine Calingo, MD Trans Team: Luces, Macaraig, Magno, Mallari

H zone - to a region consisting only of the rod-like portions of the

Topic Outline myosin molecule with no thin filaments present > M line-a region
I. Introduction to Histology where lateral connections are made between adjacent thick
filaments
II. Muscles -The lateral registration of sarcomeres in adjacent myofibrils causes
A. Types based on morphology, function and structure the entire muscle fiber to exhibit a characteristic pattern of
B. Medical Applications
transverse striations.
C. Regeneration
Thick filaments consist primarily of myosin. Myosin and actin
III. Cartilages
together represent 55% of the total protein of striated muscle while
A. Types depending on variation of ECM components the thin filaments are composed of F-actin, associated with
B. Chondrogenesis tropomyosin, which also forms a long fine polymer, and troponin, a
IV. Bones globular complex of three subunits.
A. Different bone cells
B. Bone types based on Density
C. Bone formation

I. Introduction to Histology
 Histology-study of tissues of the body and how these
tissues are arranged to constitute the organs
 Tissue- made up of cell and ECM
 Four Fundamental Tissues
1. Epithelial
2. Connective
3. Muscle
4. Nervous
 Precise combination of these tissues allow functioning
of each organ and organism as a whole
-- Optical components of a microscope:
1. Condenser – for focusing the light
2. Objective Lens – enlarges and projects the tissue sample
3. Eyepiece – magnifies the object
II. Muscles, Bones and Cartilages
A. Muscles: Responsible for contraction
Types based on morphology, function and structure:
1. Skeletal – long, cylindrical, multinucleated cells, with
cross striations, contraction is quick and voluntary
2. Cardiac – elongated and branched, with cross striations,
with intercalated disks, contraction is voluntary and
vigorous
3. Smooth – collection of fusiform/elongated cells, no
striations, slow contraction, involuntary
SKELETAL MUSCLE
- Organization: consists of muscle fibers; nuclei found
peripherally
- Muscle fibers grouped into bundles
- 3 types of covering:
1. Epimysium – covers the whole muscle tissue
2. Perimysium – covers one muscle bundle (fascicle)
3. Endomysium – covers one muscle fiber
Figure 3. Muscle bands of muscle fibers (A) Z disc, (B) M line, (C) Z
disc, (D, F) I band, (E) A band, (G) H zone, (H) sarcomere
Development of skeletal muscle:
1. Mesenchymal cells called myoblasts align and fuse together to
make myotubes.
2. Myotubes synthesize the proteins to make up myofilaments
and gradually begin to show cross striations.
3. Myotubes continue differentiating to form functional
myofilaments and the nuclei are displaced against the
sarcolemma.
4. Part of the myoblast population does not fuse and
differentiate, but remains as a group of mesenchymal cells
called muscle satellite cells located on the external surface of
muscle fibers inside the developing external lamina.
5. Satellite cells proliferate and produce new muscle fibers
following muscle injury
Four Main Muscle Proteins:
1. Actin
2. Myosin
3. Tropomyosin- each subunit is a long, thin molecule about
40 nm in length containing two polypeptide chains, which
assembles to form a long polymer located in the groove
between the two twisted actin strands
4. Troponin (TnT – responsible for moving tropomyosin
to expose actin , TnI – responsible for interaction of
actin and myosin, TnC – responsible for calcium
binding)

Figure 2. Cross section of striated muscle showing epimysium (E),

perimysium (P) and endomysium (En)
Muscle fiber - contains several parallel bundles called myofibrils
Myofibrils - consists of a long series of sarcomeres which contain
thick and thin filaments.
Z disc- separates the:
o Thin filaments - actin filaments with one end bound to -actinin,
the major protein of the Z disc. It run between and parallel to Figure 4. Molecules composing thin and thick filaments (a) a thick
the thick filaments and has one end attached to the Z line. myofilament contains 200-500 molecules of myosin (b) a thin
myofilament contains F-actin, tropomyosin and troponin
o Thick filaments - bundles of myosin, which span the entire A
band and are bound to proteins of the M line and to the Z disc
-Myosin is composed of 2 pairs of light chains and 2 pairs of
across the I bands by a very large protein called titin. They
heavy chains; contain globular projections, which have ATP
occupy the A band, the central portion of the sarcomere. binding sites and binding ability to actin
I bands - consist of the portions of the thin filaments that do not -Triad - composed of T-tubule and 2 lateral portions of
overlap the thick filaments sarcoplasmic reticulum - responsible for transporting K+
A bands - composed mainly of thick filaments in addition to >T-tubules are finger-like invaginations of the sarcolemma
overlapping portions of thin filaments

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 ANATOMY BLOCK 1

>Sarcoplasmic reticulum - regulates Ca2+ flow necessary  major fuel of the heart and are stored as triglycerides in
for rapid contraction and relaxation numerous lipid droplets seen in many cardiac muscle cells
- Lipofuscin pigment granules are often found near the nuclei of
cardiac muscle cells.

Figure 6. Longitudinal sections of cardiac muscle showing nuclei (N),

intercalated discs (I), striations (S)

Three main Junctions of Specialization within the disk

1. Fasciae
2. Maculae
3. Gap Junctions - provide continuity within the cell
 T-tubules present. larger and numerous in ventricular
muscles
 Contain numerous mitochondria (40%) - usually aerobic
metabolism
 Fatty acid - major fuel of the heart
 Lipofuscin - pigment granules, seen in old cells

Figure 5. Events of muscle contraction

General Idea:
- When Ca2+ are released in to the tissue, the Ca2+ will
bind to specific TnC → activate TnT to move → expose
the actin
- Once actin molecules are exposed the heads of the
myofilaments will attach to the actin, pulling the thin
filaments to the thick filaments
- Sarcomere will shorten → muscle contraction

In muscle contraction, the sarcomere is only shortening; there

is no decrease in length of filaments

MEDICAL APPLICATION Figure 7. Lipofuscin in cardiac muscle cells

Exercise enlarges the skeletal musculature by stimulating formation
of new myofibrils and growth in the diameter of individual muscle MEDICAL APPLICATION
fibers Ischemia - tissue damage due to lack of oxygen when coronary
 Hypertrophy - increased cell volume. arteries are occluded by heart disease.
 Hyperplasia - increase in the number of cells
 Hyperplasia takes place very readily in smooth muscle, SMOOTH MUSCLES
whose cells have not lost the capacity to divide by mitosis  Elongated, non-striated, fusiform cells and closed by a
 Myasthenia gravis - an autoimmune disorder that involves basal lamina in a network of reticular fibers
circulating antibodies against proteins of acetylcholine  connective tissues serve to combine the forces generated by
receptors. each smooth muscle fiber into a concerted action
Antibody binding to the antigenic sites interferes with acetylcholine o peristalsis – two types of smooth muscles: inner circular -
activation of their receptors, leading to intermittent periods of grinds the food eaten and outer longitudinal - move
grounded food from one place to another
skeletal muscle weakness
 Nuclei centrally located
 Ø Duchenne muscular dystrophy – a muscular dystrophy
 Found in the intestine, blood vessels and uterus
caused by defective dystrophin.
 Dystrophin is a large actin-binding protein located just
inside the sarcolemma of skeletal muscle fibers.
Mutations of the dystrophin gene can lead to defective
linkages between the cytoskeleton and the extracellular
matrix (ECM)

CARDIAC MUSCLES
- Cardiac cells form complex junction between extending
processes, with intercalated discs
- With cross striations, 1-2 centrally located nuclei
- Structure and function of the contractile proteins in cardiac
cells are essentially the same as in skeletal muscle
- T tubules: more numerous and larger in cardiac muscle than in
skeletal muscle and the sarcoplasmic reticulum is less well Figure 8. Cross section of smooth muscle in the wall of the small
intestine, cells of the inner circular (IC) layer are cut lengthwise and
developed cells of the outer longitudinal layer (OL) are cut transversely. Only
- Cardiac muscle cells contain numerous mitochondria, which some nuclei (arrows) of the latter cells are in the plane of section, so
occupy 40% or more of the cytoplasmic volume reflecting the that many cells appear to be devoid of nuclei
need for continuous aerobic metabolism in heart muscle. - A rudimentary sarcoplasmic reticulum is present in smooth
- Fatty acids: transported to cardiac muscle cells by lipoproteins muscle cells, but T tubules are not.

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 ANATOMY BLOCK 1

- The characteristic contractile activity of smooth muscle is  Capable of a more active regenerative response. After injury,
related to the structure and organization of its actin and viable smooth muscle cells undergo mitosis and replace the
myosin filaments, which do not exhibit the organization damaged tissue. Contractile pericytes from the walls of small
present in striated muscles. blood vessels participate in the repair of vascular smooth
- In smooth muscle cells, bundles of thin and thick myofilaments muscle.
crisscross obliquely through the cell, forming a lattice-like
network. Table 1. Important comparisons of the three types of muscles
- Smooth muscle actin and myosin contract by a sliding filament
mechanism similar to that in striated muscles. However,
myosin proteins are bundled differently and the cross-bridges
interact with fewer F-actin filaments.
- The thin filaments of smooth muscle cells lack troponin
complexes and instead utilize calmodulin, a calcium-binding
protein that is also involved in the contraction of non-muscle
cells.
- As in all muscle, an influx of Ca2+ is involved in initiating
contraction in smooth muscle cells. However in these cells the
Ca2+ calmodulin complex activates myosin light chain kinase
(MLCK), the enzyme that phosphorylates myosin, which is
required for myosin's interaction with F-actin.
- A number of hormones and other factors affect the activity of
Myosin Light Chain Kinase (MLCK) and thus influence the
degree of contraction of smooth muscle cells

Smooth Muscle Contraction

 Regulated by autonomic nerves, certain hormones, and
local physiological conditions such as the degree of
stretch.
 Cells occur either as multiunit smooth muscle or as
unitary smooth muscle, in which only a few cells are
innervated but all cells are interconnected by gap
junctions.
 Smooth muscle lacks neuromuscular junctions has
axonal swellings with synaptic vesicles simply lie in
close contact with the sarcolemma.
 Smooth muscle receives both adrenergic and
cholinergic nerve endings that act antagonistically,
stimulating or depressing its activity.
 Smooth muscle cells also synthesize collagen, elastin, B. Cartilages
and proteoglycans, extracellular matrix (ECM)  Composed of: chondrocytes and ECM: containing collagen,
components normally synthesized by fibroblasts glycosaminoglycans, proteoglycans that would interact
with the collagen and elastic fibers
 Function as bearing mechanical stress without distortion
 Support tissues, facilitate movements
 Have a firm, gel-like consistency
 Essential for development and growth of long bones
before and after birth
 Avascular, no lymphatics and nerves
 Covered by perichondrium - essential for growth and
maintenance of cartilage; rich in Type 1 collagen
 Found between the cartilage and tissue
 Where vascular supply and nerve can be seen

Figure 9. Diagram of smooth muscle-relaxed and contracted

MEDICAL APPLICATION
Leiomyomas - Benign tumors which commonly develop from
smooth muscle fiber but are seldom problematic. They most
frequently occur in the wall of the uterus, where they are more
commonly called fibroids and where they can become sufficiently
large to produce painful pressure and unexpected bleeding.

REGENERATION
 Skeletal muscles contain satellite cells
o Population of mononucleated spindle-shaped cells
that lies within the external lamina of each mature
muscle fiber
o After injury or certain other stimuli, the normally
quiescent satellite cells become activated, proliferating
and fusing to form new skeletal muscle fibers
 Cardiac muscle does not regenerate beyond early
childhood, lacks satellite cells
 Defects in heart muscle are generally replaced by fibroblast
proliferation and growth of connective tissue, forming
myocardial scars
 Smooth muscle also able to regenerate by mitosis
Figure 10. Diagram of cartilage transitional area between the
perichondrium and the cartilage matrix
Types depending on variations of the ECM components
1. Hyaline - most common, Type 2 collagen
 located in the following:
o articular surfaces of movable joints
o walls of nose, larynx, trachea and bronchi
o ventral ends of ribs, where they articulate with
sternum

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 ANATOMY BLOCK 1

o epiphyseal plates of long bones (makes possible
longitudinal bone growth)
o in embryo, it forms the temporary skeleton that is
gradually replaced by bone
 matrix is homogenous and glassy
o Rich in fibrils of type II collagen and aggrecan
complexes with bound water (causing matrix to
be generally basophilic in routine histology
preparations)
o Most abundant proteoglycan: Aggrecan - binds
noncovalently by link proteins to long polymers of
hyaluronic acid
 Has less collagen and more proteoglycan immediately
around the lacunae, producing slight staining
differences in the territorial matrix than that of
interterritorial matrix (where collagen is more
abundant)
 Chondronectin: binds to GAGs, collagen type II and
integrins, mediating the adherence of chondrocytes to
the ECM
→ MITOSIS & DIFFERENTIATE → Production of ECM →
Cells move further apart → Chondrocytes → DIFFERENTIATE
FURTHER → Isogenous cell aggregates (Formed groups)
o all forms of cartilage form from embryonic mesenchyme in
the process of Chondrogenesis
o two types of chondrogenesis:
a) interstitial growth - cartilaginous structures grow by
mitosis of existing chondroblasts in lacunae
 important for increasing the length of long bones
b) Appositional growth - formation of new chondroblasts
peripherally from progenitor cells in the perichondrium
 important during postnatal development
 repair or replacement of injured cartilage is very
slow and ineffective, due in part to the tissue's a
vascularity and low metabolic rate
MEDICAL APPLICATION
Calcification of hyaline matrix - a common part of aging process and
in many respects resembles endochondral ossification by which
bone is formed.

MEDICAL APPLICATION
Osteoarthritis - a chronic condition that commonly occurs during
aging
o caused by hyaline cartilage degeneration usually at joints that
are weight-bearing (knees, hips) or heavily used (wrist, fingers)
o fragments released by wear-and-tear to the articular cartilage
trigger secretion of matrix metalloproteinases which
exacerbates damage causing pain and inflammation within the
joints

2. Elastic - more pliable, Type 2 collagen and elastic fibers

o generally resembles hyaline cartilage in its chondrocytes
and major ECM components, but its matrix includes Figure 12. Chondrogenesis (a) Mesenchymal cells (b) Mitosis and
formation of chondroblasts (c) Separation of chondroblasts (d)
abundant elastic fibers, visible with special stains, which multiplication of chondroblasts form isogenous cell aggregates
increase the tissue's flexibility
o always surrounded by perichondrium Table 2. Important features of the major cartilage types
o provides flexible support for the external ear as well as
certain structures of the middle ear and larynx

3. Fibrocartilage - Coarse type 1 collagen

o Seen in body regions subjected to pulling forces
o Consists of small chondrocytes in a hyaline matrix, usually
layered with larger areas of bundled type I collagen fibers
with scattered fibroblasts
o Provides very tough, strong support at tendon insertions
and in intervertebral discs and certain other joints, as well
as in pubic symphysis
C. Bones
o Has a relative scarcity of proteoglycans which makes the - supports fleshy structures, protect the vital organs,
matrix more acidophilic reservoir for Ca2+, phosphate and other ions
o Intervertebral discs: (primarily fibrocartilages) act as - specialized connective tissue composed of intercellular
lubricated cushions and shock absorbers preventing calcified material: Bone Matrix, osteocytes, osteoblasts,
adjacent vertebrae from being damaged by abrasive osteoids
forces or impacts; has two major components - lined on both internal and external surfaces by layers of
a) Peripheral annulus fibrosis - rich in type I collagen connective tissue containing osteogenic cells: endosteum
b) Central nucleus pulpous - gel-like matrix rich in hyaluronic acid (surrounding marrow cavity) and periosteum (external surface)
- Canaliculi present - cylindrical spaces, where exchange
between osteocyte and blood capillary takes place

Figure 11. Different types of cartilage and its distribution in the body

Cartilage Formation, Growth & Repair

Chondrogenesis - production of cartilage

Mesenchymal Cells (progenitor cells) → MITOSIS &
DIFFERENTIATION → Chondroblast - immature chondroctyes

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 ANATOMY
Figure 14. Components of bone (a) section of humerus (b) parts of
compact bone (c) parts of spongy bone
Osteon - made up of one Haversian canal: Blood vessels,
lymphatic vessels, nerves
Lamellae and Lacuna (with canaliculi) - where osteon can be
seen
Major Cell Types:
1. Osteoblast (osteon + Gr. blastos, germ) - immature bone
cells, synthesize organic components of bone matrix
 when active in synthesis: cuboidal to columnar shape
and basophilic cytoplasm
 when inactive: flat and basophilia is reduced, represent
bone lining cells in both the endosteum and periosteum
 secrete components of the initial matrix, called osteoid,
that allow matrix mineralization to occur
o Basophilic - purpluish to bluish hue
o Eosinophilic - pinkish hue
2. Osteocytes (Gr. osteon, bone + kytos, cell) - found in the
lacuna, actively involved in the maintenance of bony
matrix; Death is followed by the resorption of the matrix
Figure 15. Photomicrograph of developing bone showing active
osteoblasts (Ob), osteocytes (Oc), mesenchyme (M), osteoid (Os) and
bony matrix (B)
When osteoblasts are seen in a cuboidal or columnar shape,
they are active in producing bony matrix; Inactive when they
remain flat
Osteoclasts (osteon + Gr. klastos, broken) - large, branched
motile cells; multinucleated; involved in the resorption and
remodelling of bone tissue
Figure 16. Diagram showing the relationship of osteoblasts to osteoid,
bone matrix and osteocytes
Bone Matrix - made up of organic and inorganic matters;
calcium hydroxyapatite is most abundant
- association of minerals with collagen fibers responsible for
hardness and resistance of bone tissue
- osteonectin: bone-specific mutliadhesive glycoproteins
- osteocalcin: calcium-binding glycoproteins, together with
phosphatases, promotes calcification of matrix which is
responsible for the hardness and resistance of bone tissue
- -when decalcified: bone matrix is usually acidophilic, shape of
bone is preserved but it becomes as flexible as a tendon
- Contain osteoprogenitor cells - like mesenchymal cells
Bone Types based on density
1. Compact Bone - most dense area; immediately beneath
periosteum
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BLOCK 1
2. Cancellous spongy bone - areas with numerous
interconnecting cavities deep to the compact bone
3. Long Bones – have two parts, the knobby, bulbous ends
(epiphysis) and the cylindrical part which is almost totally
composed of compact bone, with a thin region called marrow
cavity surrounded by thin region of spongy bone (diaphysis)
4. Short Bones - core of spongy bone, completely
surrounded by compact bone
5. Flat bones- have two layers of compact bone called plates,
separated by a thick layer of spongy bone called dipole
Figure 17. Thick section of bone showing the compact bone and
cancellous bone
Microscopic types:
1. Woven bone - immature bone that is formed during osteogenesis
or repair and has a calcified matrix with randomly arrange collagen
fiber.
2. Lamellar bone - most bones in adults, compact or cancellous,
characterized by multiple layers or lamellae of calcified matrix, each
3-7 mm thick
Osteon/Haversian system
 lamellae are organized concentrically around small central
canals or (Volkmann canals) containing blood vessels and
nerves
 Within each osteon, osteocytic lacunae occur between the
lamellae, with canaliculi radiating through the lamellae,
which allow all cells to communicate with the central canal.
Osteogenesis
Bone Formation
1. Intramembranous ossification - osteoblasts differentiating
directly from progenitor cells in condensed "membranes" of
mesenchyme.
2. Chondral ossification- pre-existing matrix of hyaline cartilage is
eroded and invaded by osteoblasts, which then begin osteoid
production.
Epiphyseal growth plates
-are the key to bone elongation during childhood and are
organized as an interrelated series of developing zones:
1) "Resting" zone - consists of hyaline cartilage with typical
chondrocytes
2) Proliferative zone - chondrocytes undergo mitosis and appear
stacked within elongated lacunae.
3) Hypertrophic cartilage zone - closer to the large primary
ossification center; the most mature chondrocytes in these
lacunae swell up, compress the matrix, and undergo apoptosis
4) Cartilage calcification zone - characterized by spaces created in
the matrix when they are invaded by osteoblasts, osteoclasts,
and vasculature from the primary center.
5) Ossification zone - woven bone is laid down initially by
osteoblasts and remodelled into lamellae bone
Figure 18. Osteogenesis of long bones by endochondral ossification
 ANATOMY BLOCK 1

Bone Growth, Remodelling & Repair

 Growth of bones occurs throughout life, with cells and
matrix turning over continuously through activities of
osteoblast and osteoclasts
 Lamellae and osteons are temporary structures and are
replaced and rebuilt continuously in a process of bone
remodelling by which bones change size and shape
according to changes in mechanical stress
 Bone repair after fracture or other injury involves the
activation of periosteal fibroblasts to produce an initial
soft, fibrocartilage-like callus
 Soft callus is gradually replaced by a hard callus of woven
bone that is soon remodelled to produce stronger lamellar
bone.

Metabolic Role of Bone

 Ca2+, a key ion for all cells, is stored in bone when dietary
calcium is adequate and mobilized from bone when
dietary calcium is deficient
 Maintenance of proper blood calcium levels involves
activity of all three major bone cells and is largely
regulated by subtle paracrine interaction among these and
other cells
 Hormones affecting calcium deposition and removal from
bone include parathyroid hormone (PTH), which indirectly
stimulates osteoclasts to elevate levels of calcium in blood,
and calcitonin, which can inhibit osteoclast activity,
lowering blood calcium levels

IV. References
 Junquiera’s, Basic Histology and Text Atlas, 12 and 13 ed
th th

 Wheater’s Functional Histology- A Text and Color Atlas, 6

th

ed
 Dr. Calingo’s powerpoint presentation
 http://www.education.med.nyu/Histology

V. QUIZ

1. What is the function of perichondrium?

2. What inorganic mineral salts mainly contributes to the
hardness of the bone?
3. Identify the type of bone shown in the image:

4. What is the type of bone that is generally thin and composed

of two nearly-parallel plates of compact bone tissue enclosing
a layer of spongy bone tissue like sternum and scapula?
5. How do isogenous groups form?

ANSWERS:
1. The perichondrium has vessels which provide nutrients to
the avascular cartilage. The inner layer has chondroblasts
which are responsible for the appositional growth of the
cartilage.
2. Hydroxyapatite
3. Compact bone
4. Flat bones
5. Isogenous groups form by interstitial growth of cartilage,
where the chondrocytes within the matrix divide. It is less
important than the other form of growth (appositional) from the
chondroblasts of the perichondrium.

1. T

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