Lecture 1 & 2: The Integumentary System

DLA

Integumentary System Functions:

● Barrier – physical, chemical, and biological agents

● Immunologic – antigen processing to the appropriate effector cells in the lymph system
● Homeostasis – body temp and water loss
● Endocrine – secreting hormones, cytokines, and growth factors, converting precursor molecules
into hormonally active molecules like Vitamin D
● Excretion – sweating
● Sensory – conveys info about external environment to the nervous system

Introduction: Comprises 15-20% of the body mass, varies thickness depending on position (cutis = skin)

● 2 main layers
o Epidermis – stratified squamous keratinized epithelium
o Dermis – 2 layers; loose connective tissue followed by dense irregular connective
o **the Hypodermis – layer of subcutaneous tisse
● Junction between 2 layers is serrated due to infoldings of epidermis and outpouching of dermis
● Can be classified as thick or thin based on the thickness of the epidermal layer
● Epidermal derivatives: hair follicles/hair (thick skin only), sebaceous glands (thick only), sweat
glands, nails, and mammary glands

The Epidermis: stratified squamous keratinized epithelium (5 layers) – Epidermal cells include;
keratinocytes, melanocytes, Langerhans, and Merkel

● Stratum Basale – bottom layer closest to dermis (can differentiate)

● Stratum spinosum – maturing and slowly dying cells
● Stratum granulosum – cells that are about to die
● Stratum lucidum – thick skin only
● Stratum corneum – outermost, superficial layer of dead keratinized cells

Keratinocytes – predominant cell type

● Attached to each other by desmosomes and to basal lamina by hemidesmosomes

● Sequentially move through the layers of epidermis producing keratins and lamellar bodies
o Keratins – (cytokeratins) for strength (make up 85% of cell in fully differentiated
keratinocytes
o Lamellar bodies – water barrier
● Undergo a special type of programmed cell death
● Sloughed off from surface of stratum corneum – regular exfoliation of cells from the surface of
the stratum corneum
o Regulated by proteolytic activity on the desmosomes

Melanocytes – produce melanin (neural crest derived)

 ● Basal layer – appear clear with large elongated nuclei, no desmosomal connections but are
attached to the basal lamina via hemidesmosome like structures
● Cytoplasmic processes extend between keratinocytes (1:40) (each melanocyte has about 40
keratinocyte that it interacts with)
● Retain their ability to replicate; melanocyte:keratinocyte ratio 1:4 to 1:10 regardless of skin color
● Cytoplasm stains light unless it is a melanoma (would be dark)
● Melanin transferred to surround keratinocytes for UV protection
o Skin color = amount and type of melanin present (2 types will determine shade of color)
▪ Lighter individuals and increased melanin degradation
o Oxidation of tyrosine to DOPA and then transformation into melanin
● Premelanosomes formed by Golgi and transferred into early melanosomes with gradual
conversion into melanin

Merkel’s Cells – detect touch sensations (least numerous but most abundant where sensory perception
is acute 🡪 fingertips) (sensory receptor) – very fine touch!

● Have desmosomes and contain keratin filaments, nucleus is lobed, cytoplasm contains
neurosecretory granules
● Merkel’s corpuscle – closely associated with the expanded terminal bulb of afferent myelinated
nerve fibers
● Possess antigenic markers of both epidermal and neural type

Langerhans Cell – immunologic function (encounter, process, and express antigens (Antigen Presenting
Cells – APC))

● No desmosomal junctions, nucleus stains heavily with hematoxylin (blue) and cytoplasm is clear
● Migrates to the lymph node to present the antigen to T-lymphocytes
● Involved in delayed-type hypersensitivity reactions
● TEM structure – possess tennis racquet shaped birbeck granules (small vesicles which appear as
rods with a bulbous expansion)
● Express both MHC 1 and MHC 2 and receptors for IgG

Dermis – gives strength/elasticity to skin, contains many epidermal appendages, nerve corpuscles, and
glands, contains the smooth muscle cells that move hairs, and forms a loose plexus (in reticular layer)
around erectile organs (2 layers – papillary and reticular)

● Papillary dermis layer – superficial layers (loose connective tissue)

o Contains blood vessels, predominantly type I and III collagen (elastic fibers are threadlike
and form an irregular network)
o Relatively thing and includes the substance of the dermal papillae/dermal ridges
o Contains Meissner corpuscles
● Reticular dermis layer – considerably thicker than and deep to papillary layer (dense irregular
connective tissue)
o Characterized by thick irregular bundles of mostly type I collagen and courser elastic
fibers
Epidermal/Dermal Ridges – the junction between epidermis and dermis is seen as an irregular or uneven
boundary except in very thin skin (FINGERPRINTS)

● Dermal papillae – fingerlike connective tissue protrusions (can extend into the epidermal layer)
● Epidermal Ridges (rete ridges) – infoldings from the epidermis (compliment papillae)
o Areas with increased mechanical stress have deeper ridges and longer, more closely
spaced dermal papillae

Hypodermis – layer of adipose tissue (panniculus adiposus) deep to the reticular layer of dermis

● Varies in thickness (particularly thick in cold climates)

● Serves as a major energy storage site and provides insulation
● With its associated connective tissue forms the hypodermis or subcutaneous fascia
● Hair follicles, glands, and mechanoreceptors extend into this layer

Lecture

Stratum Basale – single layer of cuboidal cells

● Mitotically active cells – keratinocyte stem cells and melanocytes

o Melanocytes mitotically divide much slower than the keratinocyte
● Merkel cells (sensory receptors)
● Less cytoplasm, more closely packed nuclei (intensely basophilic), contains many melanin
granules, provides epidermal cell renewal
● Rests on the basal lamina (attached by hemidesmosomes) with extensive cell junctions
(desmosomes)

Basal Cell Carcinoma – proliferation of basal stem cells, invades the dermis and deeper lying structures
(does not typically metastasize)

● Caused by UV light
● Dark nuclei with sparse, poorly defined cytoplasm; cells at the periphery have characteristic
palisaded appearance; central cells more randomly arranged
● May undergo ulceration
o Basal Cell carcinoma will always invade into the dermis, may undergo ulceration (dry
ulcer(scab) because there is no blood reaching the ulcer)

Bullous Pemphigoid – damage to hemidesmosomes causing separation of epithelium from dermis

(characterized by large blisters that don’t easily rupture) – Bulla = large fluid filled vesicle

● Chronic autoimmune blistering disease of skin and mucous membranes

o Antibodies (IgG) specific to hemidesmosomes bind to basement membrane and
stimulate leukocytic infiltration
o Eosinophils release proteases that degrade the hemidesmosomes
o Fluid accumulation leads to blister formation

Stratum Spinosum – several cell layers thick (larger than those in the basale)

● Gradually change their appearance as they move closer to the surface

 o Increase in size and become flattened in a plane parallel to surface
o Most noticeable in most superficial layer – nuclei become elongated
● Exhibit numerous cytoplasmic processes or spines (prickle layer)
● Cells connected to each other by desmosomes (node of bizzozero – slight thickening)
● Langerhan cells found predominantly in this layer (but are able to move freely)
o Langerhan cells have clear cytoplasm when stained

Squamous Cell Carcinoma – malignant tumor of keratinocytes and loss of orderly maturation with
variability in nuclear size and shape (hyper/parakeratosis)

● Exposure to UV with DNA damage (inactivation of p53 gene) is a cause

● Parakeratosis – outer layer of cells still have nuclei
● Common in older people (70th decade), fair-skinned > dark-skinned, and more common in
head/neck region (likely due to more sun exposure)
● Predisposing factors – sunlight (UV), industrial carcinogens, chronic ulcers, tobacco, burns,
ionizing radiation, betel nut chewing
● Islands of squamous pearl or swirls formation (differentiation)

Pemphigus Vulgaris – rare autoimmune disorder affecting epidermis and mucosal epithelium where
desmosomes break down causing stratum spinosum cells to separare

● Pemphix = blister/bubble
● Antibodies target cadherins/desmoplakins
● Separation of epidermal cell and atrophy of the prickly cell layer
● Nikolsky’s sign – skin shears by easily rubbing/wiping (easy to rupture blisters)

Stratum Granulosum – most superficial of the non-keratinized cells that varies from 1 to 3 cell layers
thick

● Contain conspicuous granules which give them granulated appearance

o Keratohyalin granules – irregularly shaped, variable size, and stain intensely basophilic
▪ Cysteine/histidine rich proteins 🡪 precursor for filaggrin

▪ Responsible for aggregating the keratin filaments within cornified cells

o Lamellar bodies – epidermal water barrier (in and out)
● Formation of epithelial water barrier – cell envelope and lipid envelope
o Cell envelope – insoluble proteins on inside of cell membrane, mechanical part
o Lipid envelope – lipid layer on the outer surface of cell membrane
▪ Lamellar bodies formed by the Golgi of spinosum cells and contents secreted by
exocytosis

Stratum Lucidum – considered a subdivision of the stratum corneum (THICK SKIN ONLY)

● Contains cells in which the keratinized process is well advanced

● Light microscopic view appears as light highly refractile eosinophilic band
Burns – Rule of 9’s used to describe burn coverage over the body and indirectly the severity (head/arms
are considered 4.5 and not 9%) – not linked to the actual percentage of skin coverage but used in order
to quickly determine the “severity” of the burn across the entire body

● 1st degree – partial thickness epidermis only, heals spontaneously

● 2nd degree – partial thickness epidermis/dermis, epithelial water barrier disrupted, heals
spontaneously
● 3rd degree – full thickness epidermis, dermis, and subcutaneous tissue, water barrier disrupted,
nerves and blood vessels destroyed, does NOT heal spontaneously, fluid loss extensive
o 2nd and 3rd degree burns almost always coincide
o 4th degree sometimes used, the burn goes through everything including soft tissues too

Stratum Corneum – several layers of dead keratinocytes with no organelles or nuclei and the deeper
portion contains the water barrier (thick plasma membrane of cells covered with an extracellular layer of
lipid)

● Filled with keratin filaments aggregated into tonofibrils, abrupt transition from granulosum
o Keratin provides the cell with strength and desmosomes keep the cells together
● Variable number of layers – Thickness of the stratum corneum will dictate if it is thick or thin skin
(basis of classification between thick/thin
o SC tends to be “not visible” in thin skin

Keratinization – total epidermal turnover time about 47 days

● Cells lose their organelles, cytoplasm, and nuclei

● More tonofilaments are formed, arranged into fibrils by filaggrin
● When reaching upper layers of SC the desmosomes are broken apart
● Cells are sloughed off

Definitions:

● Hyperkeratosis – hyperplasia of the horny (SC) layer of the skin (or the cornea)
● Parakeratosis – retention of the nuclei in the SC of the skin (psoriasis)
● Acantholysis – loss of intercellular connections resulting in loss of cohesion between
keratinocytes (pemphigus vulgaris)
● Acanthosis – epidermal hyperplasia of SS (acanthosis nigricans)

Psoriasis – a chronic inflammatory and proliferative disorder of the skin clinically manifested as
well-circumscribed, erythematous plaques covered with silvery scales (has hyper keratinization and high
turnover)

● Genetic background, inciting factors (exogenous/endogenous antigens)

● Antigen presentation by APCs, T-lymphocyte mediated immune response, secretion of cytokines
● Inflammation and cellular hyperproliferation, clinical lesions of psoriasis
o Prominent itchy areas with increased skin scaling and peeling
o Hyperplasia leads to shedding of epidermis constantly which results in scales seen as
white patches

Nails – hard plates of epidermal keratinized cells that grow by deposition of hard keratin
 ● Help with grip, gives protection (Grow 1mm per week)
● Resting on a nail bed (nail matrix and hyponysium)

Dermis – gives strength/elasticity to skin, contains many epidermal appendages, nerve corpuscles, and
glands, contains the smooth muscle cells that move hairs, and forms a loose plexus (in reticular layer)
around erectile organs (2 layers – papillary and reticular)

Epithelial Appendages: Outgrowths of the epidermis in one way or another and extend into the dermis
(Nails, Hair/hair follicle, glands)

● Hair Follicle – invagination of the epithelium that extends into the hypodermis, associated with a
sebaceous gland and smooth muscle (pilosebaceous organ)
o Sebaceous gland and hair follicle will ALWAYS be found together (pilosebaceous organ)
o Thin skin ONLY
o Consists of infundibulum, isthmus, and inferior segment
o Bulb contains the matrix cells for hair formation (melanocyte stem cells found in blub)
o Hair formation:
▪ Matrix cells contribute to formation of the internal root sheath and the hair
shaft
● Keratogenous zone: matric cells undergo keratinization as soon as they
pass through this zone
▪ Fully keratinized hair consisting of hard keratin is not followed by the internal
root sheath
● Separated from underlying dermis via thick basal lamina
▪ Nails/hair have a similar growth process with hard keratin
● Sebaceous Glands – simple branched acinar (holocrine secretion)
o Secretory portion is located in the deeper dermis, opens into hair follicle
o Product is sebum (clear with H&E stain)
o Helps prevent hair from becoming dry and brittle
● Acne – blocking of pilosebaceous organ to form blackhead or infection of tissue to cause
inflammation and pus
o Affects 85-100% of people at some point in life (affects face, back, chest, upper arm –
areas with increased sebaceous glands)
o Genetic predisposition, common in adolescents
● Eccrine Sweat Glands – simple coiled tubular glands (thick and thin skin) (merocrine secretion)
o Secretory portion is in the deep dermis, duct passes through the dermis and epidermis
to open onto the surface
▪ Composed of stratified cuboidal epithelium with basal/luminal layer
o Gentle spiral course to epidermis becomes tighter towards the surface (can see the duct
all the way through the SC layer) – tough to see duct in histo slides sometimes
o Contains clear cells (glycogen abundance), dark cells (rER/secretory granules), and
myoepithelial cells (basal aspect of the secretory segment)
● Apocrine Glands – coiled tubular glands (merocrine secretion)
 o Found only in the armpit and perineum (only found in THIN skin)
o Secretory portion is located in the dermis or even hypodermis (secretory lumen is larger
than eccrine glands) – may find these in addition to sebaceous gland
▪ Secretes pheromones
o Ducts are straight and lined by stratified cuboidal epithelium (narrow lumen) and opens
into the hair follicle

Nerve Supply – sensory receptors of various types which are peripheral terminals of sensory nerves (free
nerve ends, merkel’s corpuscle, Pacinian corpuscles, Meissner’s corpuscles, and Ruffini’s corpuscles)

● Free Nerve Endings – most numerous and terminate in the stratum granulosum
o Do NOT possess any myelin or connective tissue
o Multiple sensory modalities – fine touch, hot/cold without apparent morphological
distinction (pain/temperature)
o Networks of free dermal endings surround most hair follicles
▪ Attach to the outer root sheath, very sensitive to hair movement and serve as
mechanoreceptors (example – paper cuts, tickle, itch)
● Tickle/itch are the same sensation as pain but pain is past a certain
threshold
▪ This relationship imparts a sophisticated degree of specialization in the
receptors that surround tactile hairs as is found in the whiskers of certain
animals in which each represents its own area on the cortex
o “Sensitive to touch” is key indicator that free nerve ending is responsible
● Pacinian Corpuscle – large ovoid in deep dermis and hypodermis specialized for
pressure/vibration (looks like a cut in half onion)
o Composed of myelinated nerve ending surrounded by capsule structure, have
macroscopic dimensions more than 1mm along their long axis
▪ Nerve loses its myelin sheath after entering capsule, unmyelinated portion
covered by a series of tightly packed, flattened Schwann cell lamella that form
the inner core of corpuscle (remainder is series of concentric lamella)
● Meissner’s Corpuscle – touch receptors particularly responsive to low frequency stimuli (their
location allows for very light touch reception)
o Tapered cylinders, perpendicular to skin surface, dermal papillae just beneath the
epidermal basal lamina (appears as twisted skein of wool)
▪ At junction of dermis/epidermis (right under epidermis)
o 1 or 2 unmyelinated nerve endings of myelinated nerve fibers that follow spiral paths in
the corpuscle
● Ruffini’s Corpuscle – simplest with an elongated fusiform shape that respond to stretch and
torque
o Thin connective tissue capsule’s surrounding a fluid filled space with collagen fibers from
surrounding connective tissue passing through capsule
▪ Axonal endings are dispersed and intertwined inside the capsule
▪ Single myelinated fiber enters the capsule
 o Axonal endings respond to displacement of collagen fibers induced by sustained or
continuous mechanical stress

Wound Healing – skin damage sets in motion a sequence of events that repairs the skin to its normal (or
near normal) structure and function – 2 types: Epidermal and Deep wound healing

● Epidermal Wound Healing – occurs following wounds that affect only the epidermis (can affect
as deep as the basale layer and still be considered epidermal wound)
● In response to injury, basal stem cells of epidermis surrounding wound break contact
with basal membrane, enlarge, and migrate across wound
▪ Migrate until they meet
● Epidermal growth factor stimulates basal stem cells to divide and replace lost cells that
have moved into wound
● New cells divide to thicken the new epithelium
● Deep Wound Healing – occurs following wounds that penetrate the dermis (healing occurs in 4
phases) and healing process is more complex (scar/little loss of function involved)
● Inflammatory phase – blood clot is formed and loosely attaches the cut edges.
Vasodilation and increased permeability helps neutrophils and monocytes (which
become macrophages) to enter area and eliminate microbes, foreign material, and dying
tissue (we always have bacteria on skin and it will go into the wound). Mesenchymal
cells develop into fibroblasts
● Migratory phase – as the clot becomes a scab, epithelial cells migrate beneath it to
bridge the wound. Fibroblasts synthesize scar tissues (collagen and glycoprotein) and
damaged vessels regrow (this now constitutes the granulation tissue – important for
stitches)
● Proliferative phase – growth of the epithelial tissue at random, continued growth of
vessels
● Maturation phase – scab sloughs off, epidermis restored to normal thickness, collagen is
more organized, fibroblasts decrease and blood vessels stabilize (this type of scar is
fibrosis)
● Hypertrophic Scar – within the original wound boundary but raised more than normal (too much
“normal” collagen produced)
● Keloid Scar – excess of the boundary and extending into surrounding tissue

Skin Color – pigment produced by melanocytes consist of 2 different types (Eumelanin and Pheomelanin)
and ratio between these 2 determines skin/hair color

● Melanin is dispersed differently in dark vs. light skinned individuals and each type of melanin
appears to have a different function
o Light skin – melanin concentrated in the stratum basale
o Dark skin – pigment dispersed throughout the basale and spinosum layers
● Melanin production is influence by the surrounded keratinocytes in response to UV radiation
● Eumelanin is the darker colored pigment (tends to be congregated at stratum basale and
absorbs UV radiation)
 ● Pheomelanin is lighter and responsible for freckles (dispersed throughout and does not absorb
UV radiation)

Albinism – genetic (autosomal recessive) loss of pigmentation of the skin, hair, and eyes and the overall
loss of pigmentation can vary among individuals

● Lack of tyrosine
● Types: Ocular and Oculocutaneous
● Long term implications – skin cancers, reduce visual acuity/photophobia (macular hypoplasia),
social stigma

Vitiligo – autoimmune destruction of melanocytes that cause a depigmentation disorder

● Types: Focal, Segmental, and Generalized

● Treatment:
o Medical – topical steroid therapy, psolaren photochemotherapy, and depigmentation
o Surgical – autologous skin graft, micropigmentation, melanocyte transplant

Malignant Melanoma – malignant transformation of melanocytes (uncontrolled proliferation with

melanin being retained inside the cell), increased number of melanocytes with large atypical morphology
arranged at the dermo-epidermal junction (may invade the dermis)

● Metastases – fatal, causes highest number of skin cancer related deaths in the US
● More common in whites, Australia/South Africa from exposure to UV sunlight (acute,
intermittent, and blistering)
● Asymmetry, Border (irregularity/raised), Color (differences in same area), Diameter (1/4 inch or
6mm), Evolving (change over time/different parts changing of same lesion)
● Eumelanin may be protective of MM by absorbing UV radiation, pheomelanin could potentially
promote MM
o Seems to be a correlation to increased amount of Pheomelanin and predisposition to
skin cancer
● Melanin A – immunohistochemistry marker, a specific antibody that stains malignant melanoma
cells
Lecture 7: Development of the Nervous System
DLA – Early Embryology

Week 1: Fertilization

● Capacitation – removal of seminal proteins from the surface, penetration through the corona
radiata
● Acrosome Reaction – release of acrosomal enzymes, penetration of sperm through zona
pellucida
● Fusion of plasma membranes or sperm/oocyte – changes in zona pellucida inactivates sperm
receptors and prevents polyspermy
● Fertilization stimulates completion of 2nd meiotic division to form the mature oocyte
● Nucleus = female pronucleus, sperm nucleus forms the male pronucleus then fusion of pronuclei
o Restores normal diploid number to make zygote (embryonic sex is determined)
● DNA replication, mitotic division and cleavage begins

Week 1: Cleavage, Morula, and early Blastocyst

● Cleavage – repeated mitotic division of the zygote to produce identical daughter cells or
blastomeres
● Zygote is subdivided without increase in size
● After 16-32 blastomeres 🡪 morula (day 4)

Week 1: Blastocyst Formation (days 4-6)

● After the morula enters the uterus a fluid-filled space, the blastocyst cavity appears inside the
morula. The fluid passes from the uterine cavity through the zona pellucida to form this space.
As fluid increases in the blastocyst cavity, it separates the blastomeres into two parts
o Centrally located cells (or inner cell mass) 🡪 embryoblast 🡪 embryo (located at the
embryonic pole
o Peripheral cells 🡪 trophoblast 🡪 fetal component of the placenta

Week 1/2: Implantation (normal site – posterior wall of body of uterus, close to fundus)

● Approximately 6 days after fertilization the blastocyst attaches to the epithelium of the
endometrium (inner lining of the uterus), usually adjacent to the embryonic pole
● As soon as the blastocyst attaches to the endometrial epithelium, the trophoblast proliferates
rapidly and differentiates into two layers; cytotrophoblast (inner layer) and syncytiotrophoblast
(outer layer)

Week 2: Implantation (2 distinct layers of the trophoblast)

● Inner cytotrophoblast makes cells (mitotically active, contributes cells to the

syncytiotrophoblast)
● Outer syncytiotrophoblast for secretion (secretes Human Chorionic Gonadotrophin or HCG)
 o mass of cells or syncytium at the embryonic pole erode endometrial connective tissue
enabling the blastocyst to “burrow” into the endometrium

Ectopic Pregnancy: abnormal sites of implantation

● signs/symptoms – missed period, abdominal pain, lower than normal HCG

● causes – factors that delay or prevent transport
● sites – 95-98% occur in the uterine tubes (most often in ampulla and isthmus)
● sequelae – tubal rupture and hemorrhage
● treatment – termination of pregnancy, possible removal of tube

Week 2: Bilaminar Disc

● As implantation of the blastocyst progresses, a small space appears in the embryoblast, which is
the primordium of the amniotic cavity
o amniogenic (amnion forming) cells, amnioblasts, separate from the epiblast and form
the amnion
● Concurrently, morphologic changes occur in the embryoblast (cluster of cells from which the
embryo develops) that result in the formation of a flat, almost circular bilaminar plate of cells,
the embryonic disc, consisting of two layers
o Epiblast – thicker layer consisting of high columnar cells related to amniotic activity
▪ forms the floor of the amniotic cavity and is continuous peripherally with the
amnion
o Hypoblast – consisting of small cuboidal cells adjacent to the exocoelomic cavity
▪ forms the roof of the exocoelomic cavity and is continuous with the thin
exocoelomic membrane
● the membrane together with the hypoblast lines the primary umbilical vesicle
● The embryonic disc now lies between the amniotic cavity and primary umbilical vesicle
● Cells from the vesicle endoderm form a layer of connective tissue, the extraembryonic
mesoderm, which surrounds the amnion and umbilical vesicle

Week 3: Gastrulation (bilaminar disc 🡪 trilaminar) – the cells of the epiblast, through the process of
gastrulation, give rise to all three germ layers in the embryo, the primordia of all its tissues and organs

● Primitive streak - beginning of the third week, a thickened linear band of epiblast appears
caudally in the median plane of the dorsal aspect of the embryonic
o Left/right, caudal/cranial. And dorsal/ventral can now be identified
o Cranial end proliferates for form primitive node
o Concurrently, a narrow groove, the primitive groove, develops in the primitive streak
that is continuous with a small depression in the primitive node, the primitive pit
● First wave of epiblast cells displaces the hypoblast to form endoderm
● Epiblast cells migrate laterally between new endoderm and epiblast to form intraembryonic
mesoderm
● Remaining epiblast constitutes the ectoderm (which will later differentiate into nervous tissue)
Sacrococcygeal Teratoma: most common tumor in newborn infants from persistence of remnants of the
pluripotent primitive streak in the caudal region of the embryo

● More common in female infants

● Derivatives from all 3 germ layers may be found (teether, hair, muscle, glands, etc)

Week 3: Notochord Formation – the mesodermal cells at the primitive node migrate cranially from the
primitive node and pit, forming a median cellular cord, the notochordal process

● Process soon acquires a lumen, the notochordal canal. The notochordal process grows cranially
between the ectoderm and endoderm until it reaches the prechordal plate
● Ectoderm and endoderm remain firmly adhered to each other at two points in the embryonic
disc
o At the prechordal plate where oropharyngeal membrane forms
o At the cloacal membrane which is caudal to origin of the primitive streak
● Notochord becomes detached from the endoderm of the umbilical vesicle (umbilical vesicle once
again becomes continuous layer)
o Adult remnants of notochord are nucleus pulposus of discs and apical ligament from
dens to occiput
o Persistence of additional remnants may result in a chordoma, a slowly growing
malignant tumor which invades bone (occurs at base of skull or lumbosacral region)
● Fate of mesoderm 🡪 notochord, cells which migrate around prechordal plate (cardiogenic
mesoderm), and paraxial, intermediate, and lateral plate mesoderm
o Paraxial mesoderm – adjacent to the developing notochord and neural tube
▪ Head – forms loose mesenchyme (skeletal muscles of face, jaw, and throat)

▪ Somites – paired segmented bodies on either side of notochord in future trunk

and neck region
● Somites form most of the axial skeleton, dermis, voluntary muscles of
the head, limbs, and body wall (3 main derivatives)
o Dermatome = dermis (fibroblasts)
o Sclerotome = vertebrae and ribs
o Myotome = primordial muscle cells
▪ Dorsal Epiaxial – extensors of the neck, vertebral
column, and lumbar region (sacrococcygeal ligament)
▪ Ventral Hypaxial – limbs, abdominal wall, intercostal,
scalene, prevertebral, infrahyoid, and geniohyoid mm,
respiratory/pelvic diaphragm
o Intermediate Mesoderm forms the gonads and urinary system
o Lateral Mesoderm – clefts appear in lateral plate mesoderm which coalesce to form
intraembryonic coelom (IC) splitting into 2 layers
▪ Parietal (somatopleure) Layer – formation of the body wall
 ▪ Visceral (splanchnopleure) Layer – formation of remained of organs

Week 4: Folding – there is rapid growth of the embryo, the now trilaminar embryo undergoes folding
and adopts a cylindrical shape a 3-D body plan (tube within a tube)

● Outer Tube = ectoderm; Inner Tube = endoderm (both tubes separated by a mesodermal layer)
● Folding of the embryo is due to differential growth of various embryonic structures and results in
the embryo being separated from the extraembryonic structures
● Occurs in 2 planes – lateral folding = cylindrical disc; cephalocaudal folding = growth of the
neural tube causing head and tail folds

Lecture

Neurulation – the induction of the ectoderm to form the primordium of the entire central nervous
system

● Structures which developed during week 3 (notochord and paraxial mesoderm) induce overlying
ectoderm to differentiate into neuroectoderm
● Ectodermal cells immediately dorsal to the notochord become elongated and form a thickened
single layered neural plate
● A longitudinal median neural groove appears around day 18 with a neural fold on each side

Process of Neurulation:

● By the end of the 3rd week the neural plate consists of a wide cranial portion→ future brain and
a narrow caudal portion→ future spinal cord
● As somites continue to develop and be added at the trunk region the future spinal cord region of
the neural plate is forced to lengthen
● Eventually the neural folds approximate and begin to fuse closing the neural groove (Starts on
day 22 at the occipital and cervical region)
o Extraembryonic mesoderm forms on the outside
● As the tube fuses the neuroectodermal cells along the crest of the neural folds separate
o Neural crest cells from the junction of surface ectoderm and neuroectoderm pinch off
and come to lie along the sides of the neural tube
● The closure of the tube progresses both cranially and caudally
o The neural tube caudal to the 4th pair of somites will form the spinal cord
● A narrow neural canal is now present which gets smaller as the cells in the wall of neural tube
proliferate
o the ends of tube initially remain open to the amniotic cavity - these openings are the
rostral (closes day 25) and caudal (closes day 27) neuropores
▪ whatever day the rostral closes then caudal will close 1-2 days later (doesn’t
always close on day 25 then day 27)
● Surface ectoderm again becomes continuous across dorsal surface and neural tube migrates
below surface

Spinal Cord Development:

 ● Neural tube is lined by thick layer of neuroepithelium which forms the ventricular zone
o The marginal zone forms the layer superficial to ventricular zone
● Cells in the ventricular zone multiple and the newly formed cells migrate to form the
intermediate zone (mantle layer) between the ventricular/marginal zone
o Cells in the intermediate zone differentiate into neuroblasts which will form neurons
● The intermediate zone thickens into 4 regions
o Pair of alar plates – dorsal horns and dorsal grey columns
o Pair of basal plates – ventral somatic/lateral horns
o The sulcus limitans separates the alar and basal plates
● The intermediate layer forms spinal grey matter
● Marginal layer forms white matter
● Cavity of the neural tube forms the central canal
● Ventricular zone forms ependymal lining of the central canal

Neural Tube Derivatives: Mesenchyme and Neuroepithelium (cells of CNS come from 2 sources)

● CNS – brain, brain stem, spinal cord

● PNS – somatic motor portion, preganglionic visceral motor portion
● Mesenchyme 🡪 mesenchymal cell 🡪 microglial cell (come from bone marrow)
● Neuroepithelium leads to neurons, macroglial cells, and epithelium of choroid plexus

Neural Crest Derivatives: Neural Crest Cells (very diverse)

● Ganglion cells of the PNS, Sensory Ganglia of cranial nerves, Para/Sympathetic Ganglion
● Melanocytes (in skin)
● Spinal Ganglion (DRG) 🡪 unipolar neuron 🡪 Schwann Cell, Multipolar Neurons
● Ganglion of Sympathetic Truck 🡪 Celiac Ganglion, Renal Ganglion, Suprarenal Medulla
(Chromaffin cells)
● Leptomeninges (Pia/Arachnoid)

Primordial Meninges:

● Adjacent mesenchyme condenses to form a primordial meninx with two layers

o Inner – derived from neural crest is leptomeninges (pia/arachnoid)
o Outer – derived from mesoderm is dura mater

Myelination – starts during late fetal period and continues through first year of life

● PNS – myelin is formed by Schwann Cells (from neural crest)

o 1 Schwann cell myelinates 1 neuron
● CNS – myelin in brain/spine formed by Oligodendrocytes (from neural tube)
o 1 oligodendrocyte can myelinate multiple neurons

Formation of the Vertebral Column (3rd week)

● Intervertebral Disc – this develops from the caudal dense part of the Sclerotome and forms the
anulus fibrosus of the disc
 o Each vertebra develops from mesenchyme of the pair sclerotomes which surround the
neural tube and the notochord at each level
o Sclerotome has two regions: densely and loosely packed cells
▪ Dense – forms IV disc and part of vertebral body

▪ Loose – forms vertebral body together with the rest of dense tissue
o Hemi-vertebra occurs if one of the pairs of sclerotomes fail to develop
● Chordoma – rare malignant tumor formed when remnants of the notochord persist, may
infiltrate bone and occur most frequently at the base of the skull
o The notochord is the region between developing vertebral bodies persists and forms the
nucleus pulposus
● Vertebral Column – the remnants of the sclerotome form the mesenchymal primordium of the
vertebra
o Appears at the 5th week begins to convert to cartilage and later bone by the end of the
7th week when the first primary ossification centers appear
o 2 ossification centers: centrum (body) and 2 neural arch centers
▪ Neural arch (future vertebral arch) is 1st to ossify
● Typical Vertebra – at birth the typical vertebra is made up of three bony parts joined by
cartilaginous joints (bones fuse between ages 3 to 6)
o Ossification continues until the 25th year (5 secondary ossification centers appear after
puberty and unite by the 25th year)
o At the secondary ossification centers in the transverse processes the costal elements
form ribs in thoracic region, mammillary processes in lumbar region, and cervical rib the
costal elements form ribs
● Atypical Vertebra
o C1 and C2 centra of C1/C2 fuse forming the dens, no IV disc formed
o Sacrum transverse and costal elements fuse
o Coccyx

Clinical Considerations: Neural tube Defects; genetic and environmental factors are thought to be
involved in the production of NTD’s and some medications like anticonvulsants and valproic acid may
also cause NTD’S (Prophylaxis with folic acid is recommended for all women of reproductive age)

● Spina Bifida – varying degrees of failure of neural tube formation involving the caudal neuropore
with accompanying defects in formation of meninges and neural arch of vertebra
o Spina Bifida Occulta – no bulge over the bony defect, no neurological deficits, a tuft of
hairs or skin dimple may be present (most common in the lumbosacral area)
o Spina Bifida Cystica with Meningocele – bulge is seen over the defect, contains dura,
meninges, and CSF in the subarachnoid space (cyst in lumbar region), neurological
deficits relatively uncommon
o Spina Bifida Cystica with Meningomyelocele – bulge is seen over the defect, contains
meninges, cord, nerves, and vertebral bodies (membranous sac can be observed filled
with CSF)
 ▪ Remnants of neural tissue can also be observed

▪ Severe neurological deficits are common ranging from paralysis, sensory loss,
and bladder dysfunction
o Spina Bifida Cystica with Myeloschisis – skin and bony defect with open spinal cord,
seen as a mass of neural tissue (usually not compatible with life)
● Meroencephaly – failure of rostral neuropore to close leading to absence of a large part of brain
and skull
o Usually embryos will not make it to full term (miscarriage) and if born they do not live
for long (only days)
o Exact cause is unknown (environmental toxins, lack of folic acid)

Lecture 5: Vertebral Column, Spinal Cord, Nerves and Plexus – Part 1

Lecture

Vertebral Column – 7 cervical, 12 thoracic, 5 lumbar, 5 sacral (fused), and 3-5 coccygeal (fused)

● Atlas (C1) and Axis (C2) – no intervertebral disc between C1 and C2

o Dens (odontoid process) acts as a pivot that allows the atlas and head to rotate
side-to-side
● Typical vertebra – body, arch (lamina and pedicle), and spinous process
● Cervical, thoracic, and lumbar all have unique shape
o Cervical – bifid spinous process and transverse foramen (C7 spinous process is longest in
the body)
o Thoracic – long spinous process, facets for rib articulation
o Lumbar – larger body (for more weight), mamillary process
● Atypical vertebrae – sacrum and coccyx (fused) with transverse ridges

Ligamentous Support of Spine – 5 spinal ligaments

● Anterior Longitudinal Ligament – extends along the longitudinal (up and down) anterior surface
of the spinal column (thick at the top)
● Posterior Longitudinal Ligament – extends along the longitudinal (up and down) posterior
surface of the spinal column (thick at the bottom)
● Ligament Flavum – connects all of the lamina
● Supraspinous Ligament – connects all of the spinous processes posteriorly
● Interspinous Ligament – connects all of the spinous processes to each other

Ligaments of the Atlas/Axis Connection:

● Cruciate ligament – hold dens in place during rotation of the head

● Apical ligament – deep to the cruciate ligament, attaching dens to the skull (occipital bone)
● Alar ligaments – laterally from dens to skull
 ● **There are 3 Atlanto-axial joints. Movement at all three Atlanto-axial joints permits the head to
be turned from side to side (no movement). During this movement, the cranium and C1 rotate
on C2 as a unit. During rotation of the head, the dens of C2 is the axis or pivot that is held in a
socket formed anteriorly by the anterior arch of the atlas and posteriorly by the transverse
ligament of the atlas
o Transverse ligament is part of the cruciate ligament – thick, strong bad, which arches
across the ring of the atlas and retains the odontoid process in contact with the atlas
● **Vertically oriented but much weaker superior and inferior longitudinal bands pass from the
transverse ligament to the occipital bone superiorly and to the body of C2 inferiorly

Transverse Ligament Rupture – destabilizing injury that can occur in isolation or with atlantoaxial
subluxation or an atlas fracture KNOW THIS SLIDE

● Primary restraint to anterior translation of atlas in relation to the lower cervical spine
● Can fail in midsubstance or avulsion of bone can occur
● Estimated that a force of about 85kg is required to rupture the transverse ligament

Intervertebral Disc – cartilaginous joint between 2 adjacent spinal discs (nucleus pulposus and anulus
fibrosis) that allows lateral movement

● Herniated disc – anulus rupture will cause nucleus to come posterior and lateral

Spine Curvature:

● Primary Curvatures – develop during the fetal period in relationship to the flexed fetal position
● Secondary Curvatures – result from extension from the flexed fetal position and maintained by
the differences in thickness of between anterior and posterior parts of IV discs (cervical (when
child is able to lift head) and lumbar (when child is able to start walking) portions of spine)
● Kyphosis – hunchback, abnormal increase in the thoracic curvature and as a result the vertebral
column curves posteriorly
● Lordosis – characterized by an anterior tilting of the pelvis (the upper sacrum is flexed or rotated
antero-inferiorly) with increased extension of the lumbar vertebrae, producing an abnormal
increase in the lumbar kyphosis – seen in pregnant females from baby weight
● Scoliosis – characterized by an abnormal lateral curvature that is accompanied by rotation of the
vertebrae (common in young adults)

Spinal Cord – conduction pathway between body and brain (part of the CNS)

● Protected by Vertebral column, meninges, and CSF

● Gives rise to 31 spinal nerves (8 cervical, 12 thoracic, 5 lumbar, 5 sacral, and 1 coccygeal)
● 2 enlargements – cervical and lumbosacral (thickened spinal cord in these regions)
● Tapers off at the conus medularis (at L1/L2)
● Cauda Equina (dorsal and ventral roots) – continue into the lumbar cistern

Spinal Meninges – protective membranes covering the spinal cord and continuous with cranial meninges
covering the brain

● Dura Mater: outer covering, dense connective tissue (tough outer covering)
o Vascular, innervated collagen fibers
 o Sub-dura space below the mater (only time you will find a space here is with a
pathology)
o Epidural mean above the dural space (has nerve roots, loose fatty tissue, small arteries,
and internal vertebral venous plexus of Batson)
● Arachnoid mater: avascular, loose connective tissue (middle) – spider-web like appearance
o Lined back to back by simple squamous epithelium
o Sub-arachnoid space below the mater (contains CSF)
● Pia Mater: vascular loose connective tissue (inner, delicate tissue)
o Lies directly on brain and spinal cord
o Sub-pia space below the mater

Denticulate Ligaments – (extension of pia mater) 21 pairs composed of a single narrow fibrous strip that
extends from the craniovertebral junction to T12, each ligament features 18-20 triangular extensions of
pia mater that attach to the dura at their apices (Separate the ventral and dorsal rootlets from each
other) – stabilizes the spinal cord

Supraspinous ligament > interspinous ligament > ligament flava > epidural space > dura mater > sub-dura
space > arachnoid mater > subarachnoid space containing CSF (from superficial to deep)

● From midpoint, from lateral you would hit interspinous ligament and then everything else (you
miss supraspinous ligament)

Caudal end of spinal cord changes position through development: 8 weeks > 24 weeks > newborn > adult
(in adults the spinal cord ends at L1/L2 and has reached here by about age 2)

● Dural sac extends from L1/L2 to S2 (cannot do lumbar puncture lower than S2)

Spinal Arteries: blood supply to the cord

● Artery of Adamkiewicz – important in circulation to lumbar and sacral parts of cord

● Vertebral A., Anterior Spinal A., Posterior Intercostal A., Lumbar & Sacral A.
● The spinal cord may suffer circulatory impairment if the segmental medullary arteries and
particularly the great anterior segmental medullary artery (of Adamkiewicz) undergo
Atherosclerosis, aorta clamping, aortic aneurysm, etc
o Blocking of certain arteries can be overcome at different levels of the spine by other
arteries

Venous Drainage: internal and external venous plexus (of Batson) – VALVELESS

● Can serve as collateral pathway for venous flow to systemic veins (caval and azygos veins)
● No checkpoints so cancer can metastasize up to cranium “unchecked”
● Internal bleeding drains into epidural space

Lumbar Puncture: The skin covering the lower lumbar vertebrae is anesthetized and a lumbar puncture
needle is inserted in the midline between the spinous processes of L4 and L5 at the supracristal line
(dotted line). After 4-6cm the needle “pops” through the ligamentum flavum and then “pops” again
when penetrates the dura to enter the lumbar cistern (enters dural sac with CSF)

● Is NOT performed in the presence of increased intracranial pressure

 ● Needle into subarachnoid space to drain CSF
● Needle is directed cephalad (headwards)

Lumbar Epidural Anesthesia: epidural steroid injection is injected into the epidural space

Sacral/Caudal Epidural Anesthesia: Provides a reliable and effective block for operations which involve
low lumbar and sacral dermatomes

● Often used for steroid injections

● Landmark – sacral hiatus
● Decreased risk of entering subarachnoid space

Lecture 6: Vertebral Column, Spinal Cord, Nerves, and Plexus – Part 2

Lecture

Nervous System Overview: CNS and PNS

● Somatic Sensory/Afferent – receptors in body wall for conscious sensation (pain, pressure, touch,
temp, proprioception)
● Somatic Motor/Efferent – voluntary control of skeletal muscle in body wall
● Visceral Sensory/Afferent – receptors in organs and tissues (distension, inflammation, ischemia,
physiological changes, abnormal muscle spasms)
● Visceral Motor/Efferent (autonomic) – involuntary control of smooth and cardiac muscle and
glands

Spinal Cord Organization – External:

● 31 pairs of spinal nerves (8 cervical, 12 thoracic, 5 lumbar, 5 sacral, and 1 coccygeal)

● Spinal cord ends at vertebral levels L1/L2
o Conus medullaris (end of cord)
o Cauda Equina (below conus medullaris – spinal nerve root descending to exit as lumbar
and sacral nerves)

Spinal Cord Organization – Internal:

● 2 types of tissue:
o Inner gray matter – cell bodies of neurons and glial cells (support cells)
o Outer white matter – myelinated axons (neural highway)
● Dorsal Horn (sensory), Ventral Horn (motor), and Lateral Horn (cell bodies of preganglionic
visceral sympathetic – motor)
● Lateral Horn – is primarily involved with activity in the sympathetic division of the autonomic
motor system
o Composed of sympathetic preganglionic visceral motor neurons
 o Present at 17 levels of the spinal cord, specifically through levels T1-L2 (cell body of
preganglionic sympathetic outflow) as well as levels S2-S4 (cell body of preganglionic
parasympathetic outflow)
▪ ONLY found from T1-L2 and from S2-S4

Spinal Nerve Formation:

● Dorsal/Ventral rootlets > dorsal/ventral root > spinal nerve (mixed) > dorsal/ventral ramus
(mixed)
o Dorsal – presence of Dorsal Root ganglion (cell bodies) between the dorsal root and
spinal nerve (sensory coming back to spine)
o Ventral – motor neurons going out of spine to rest of body
o Grey Ramus Comm. – sympathetic system present near ventral ramus (incoming)
o White Ramus Comm. – sympathetic system present near ventral ramus (outgoing)
▪ Sympathetic chain ganglion

● Spinal nerve exits through intervertebral foramen

o C1 – exits between skull and C1 vertebra (above same number vertebra)
o C2-C7 – exit above same number vertebra
o C8 – exit above T1 vertebra or below C7 (acts as C8 since no C8 vertebra exits)
o T1-end of cord – exit from spinal canal through intervertebral foramen below the same
number vertebra
o L3-L5 – exits right below pedicle and above intervertebral disc (large space since large
vertabra)
▪ Herniate disc cause posterior and lateral movement due to immediate posterior
path blocked by posterior longitudinal ligament
▪ Compressed nerves in this region are the nerve below the IV disc (example.
Compressed nerve between L3-L4 would be the L4 nerve NOT the L3 nerve)
● This WILL be on the test
● Dermatomes – an area of skin innervated by a single spinal nerve (because of overlap between
adjacent spinal nerves, sensory (cutaneous) loss results only if 2 or more consecutive spinal
nerves are lesioned) – KNOW THE DERMATOME MAP (as these are landmarks to know for
patient)
o C5 – shoulder
o C6 – thumb
o C8 – pinkie
o T4 – nipple
o T7 – tip of xiphoid process
o T9 – affected by Herpes Zoster (shingles)
o T10 – umbilicus
o L1 – inguinal region
o L4 – great toe
o S1 – little toe
 o S2,3,4 – perineum

Innervation Routes: Spinal Nerve and its branches contain mixed fibers (sensory/motor/autonomic)

● Somatic Motor Neuron - Motor Neuroblast (from basal plate of intermediate layer)
o Muscles of the back (dorsal ramus)
o Muscles of the body wall and limbs (ventral ramus)
● Visceral Motor Neuron – motor neuroblast (from basal plate of intermediate layer) forming
pre-ganglionic sympathetic going to motor neuroblast (from neural crest) forming
post-ganglionic sympathetic
o Both dorsal and ventral ramus
● Primary Sensory Neuron – sensory neuroblasts (from neural crest) in DRG
o Skin of the back (dorsal ramus)
o Skin of the body wall and limbs (ventral ramus)

Myotomes:

● Regions of skeletal muscle innervated by a single nerve or spinal cord level

● Most individual muscles are innervated by more than 1 spinal cord level
● Myotome testing is accomplished by testing movements of joints or muscle groups

Organization of Nerves: Plexus – a group a nerves

● Somatic Plexus (body wall structures) – a network of nerves derived totally from ventral rami of
spinal nerves (motor fibers > muscle and sensory fibers > skin/joints)
o Somatic Nerve Plexi – cervical, brachial, lumbar, and sacral plexus’
▪ Branches of a somatic nerve plexi are somatic nerves (median nerve, ulnar
nerve, pudendal nerve, etc)
o Hilton’s Law – nerve crossing a joint, innervated that joint (example – median nerve
crosses elbow joint and can thus innervate that joint)
● Visceral Plexus (organs and glands) – carries autonomic nerve fibers (para/sympathetic) to
smooth and cardiac muscle and glands, also carries sensory fibers back from these areas

Summary:

● Dorsal (posterior) ramus – supplies intrinsic back muscles (motor) and skin over that area
(sensory)
● Ventral (anterior) ramus – supplies all other muscle (motor) and skin over area (sensory)
● Sympathetic Chain ganglia – communicate with spinal nerve via grey (unmyelinated) and white
(myelinated) rami communicantes
o SCG run down length of vertebral column lateral to vertebral bodies (bilateral)

DLA

General Characteristics of the ANS:

● Visceral – innervates 3 target tissues (smooth muscle, cardiac muscle, and glands)
● Motor – voluntary skeletal muscle control
 ● Sensory fibers travel along with autonomic nerve fibers
● 2 major divisions: Sympathetic (“fight-or-flight”) and Parasympathetic (“Rest-and-digest”)
o Enteric (minor 3rd component that is gut-related
● Sympathetic Functions: (catabolic) pupillary dilation, ventilation dilation for increased breathing,
increased blood flow to skeletal muscle, increased HR, force of cardiac contraction, and cardiac
output, increased arterial pressure
● Parasympathetic Functions: (homeostatic/anabolic) decreased BP, decreased HR, constricted
bronchial/respiratory tree for decreased breathing rate, stimulation of gut activities, warm skin,
pupillary constriction (for close site)

Neuron Pathways:

● Somatic Motor Neurons – one long, heavily myelinated axon extending from CNS to effector
● Autonomic Motor Neurons – 2 neuron system, with a lightly myelinated preganglionic neuron
with cell body in the CNS and an unmyelinated postganglionic neuron with the cell body in the
autonomic ganglion in the PNS
o Sympathetic – short preganglionic, long postganglionic (T1 – L2)
o Parasympathetic – long preganglionic, short postganglionic (cranial nerves III, VII, IX, X
and S2-S4)
● Sympathetic Postganglionic Locations:
o Sympathetic Chain (Trunk) – lateral aspect of vertebrae from superior cervical ganglion
at the base of skull to the ganglion impar at the level of the coccyx
o Associated with each spinal nerve: 1 pair per spinal nerve
● Parasympathetic Postganglionic Locations:
o Found scattered near or in the walls of target organs
o Discrete ganglia are only found in the head associated with the parasympathetic cranial
nerves

Sympathetic Route: Lateral Horn T1-L2 > Ventral Root > Spinal nerve/ventral rami > WRC > sympathetic
chain (after chain can go to body wall and limbs, viscera, or organs/structures in the head)

● Once the fibers enter the sympathetic chain (SC) they can;
o Synapse immediately with post-ganglionic neuron at the same paravertebral level
o Ascend in the SC to synapse with a post-ganglionic neuron at a higher paravertebral level
o Descend in the SC to synapse with a post-ganglionic neuron at a lower paravertebral
level
o Pass through the SC without synapsing to continue as a splanchnic nerve and synapse at
the pre-aortic ganglia
● Sympathetic to body wall and limbs 🡪 targets are sweat glands, vascular smooth muscle (skin,
connective tissue, and skeletal muscle), and arrector pilli muscles
o Sympathetic to spinal nerves at T1-L2
▪ Preganglionic axons from cell bodies in lateral horn of spinal segments T1-L2
leave the cord via ventral roots
▪ Enter the SC via WRCs
 ▪ Synapse in ganglia at the same spinal level
▪ Postganglionic exit the chain via GRC to join spinal nerves T1-L2
▪ Dorsal/ventral rami of these spinal nerves innervate the body wall at T1-L2
o Sympathetic supply to spinal nerves above T1
▪ Preganglionic axons from cell bodies in lateral horn of upper thoracic segments
leave the cord via ventral roots
▪ Enter the SC via WRCs
▪ Ascend in chain
▪ Synapse in cervical ganglia
▪ Postganglionic exit the chain via GRC to follow spinal nerves C1-C8
▪ Dorsal/ventral rami of these spinal nerves innervate neck/upper limbs above T1
o Sympathetic supply to spinal nerves below L2
▪ Preganglionic axons from cell bodies in lateral horn of lower thoracic and upper
lumbar spinal segments leave the cord via ventral roots
▪ Enter the SC via WRCs
▪ Descend in chain
▪ Synapse in lower lumbar and sacral chain ganglia
▪ Postganglionic exit the chain via GRC to join spinal nerves L3-S5
▪ Dorsal/ventral rami of these spinal nerves innervate the body wall below L2 and
lower limbs
● Sacral Parasympathetic:
o Preganglionic cell bodies in small “lateral-horn-like area” of sacral spinal cord (S2-S4)
o Axons travel on ventral roots to spinal nerves and then ventral rami of S2-S4
o Leave ventral rami as pelvic splanchnic nerves to enter the pelvic plexus
o Postganglionic nerves are located at or in walls of target organs in the pelvis (such as
urinary bladder)
● Visceral Sensory Fibers (Visceral Afferents):
o Travel along with fibers from the PNS/SNS back to the CNS
o Cell bodies of VA found in the DRG of the spinal segment where the sympathetic and
parasympathetic stimulus leaves
o VA traveling with para/sympathetic:
▪ Parasympathetic – mediate unconscious sensations such as atrial or GI stretch
and results in involuntary physiological responses such as enhanced peristalsis,
decreased HR and BP, etc
▪ Sympathetic – mediate conscious sensations that are usually painful and
respond to ischemia, distention (abnormal stretch), inflammation, or spasmodic
contraction of smooth muscle

Referred Pain: Pain being interpreted as coming from another area of sensory input

● Occasionally, sensory information that enters the spinal cord from one location can be
interpreted by the CNS as coming from another location that is innervated by the same spinal
cord level
 ● Can occur when information coming from the viscera (an area of low sensory output) enters the
spinal cord at the same level as information coming from the body wall (an area of high sensory
output)
o This can result in the pain being interpreted as coming from the area of high sensory
output

Lecture 3: Cartilage
Lecture

Cartilage – specialized type of connective tissue (unique cells, fiber population, and ground substance)
that confers shape, flexibility, elasticity, shock absorbance, tensile strength, and a model for long bone
formation

● 95% is extra cellular matrix – solid and firm but pliable (accounts for its strength/resilience)
● 5% is cells – chondrocytes and chondroblasts
● Major component of the ground substance is GAGs – highly hydrophilic to trap water molecules
and give a “squishy” texture

Hyaline Cartilage – locations include; nose, tracheal/bronchial rings, laryngeal cartilages, costal
cartilages, articular surface of long bones, epiphyseal growth plate, and fetal skeleton – most abundant
cartilage in the human body

● Histogenesis – mesenchymal cells; contains chondroblasts and chondrocytes cells and ECM is
Type II collagen fibers, GAGs, and glycoproteins
o Mesenchymal > chondrogenic cells > chondroblasts > chondrocyte cells
● Growth – appositional & interstitial; degeneration – does not readily degenerate (fairly
resistance throughout life); calcification – does calcify, bone formation and aging (calcified
cartilage is 1 step away from bone formation)
● AVASCULAR – nutrients supplied by vessels from perichondrium by diffusion
● Perichondrium – connective tissue covering entire cartilage (except articular surfaces) consisting
of 2 layers
 o Outer fibrous layer (FL): contains fibroblasts (synthesize collagen type 1 and matrix),
blood vessels
o Inner cellular layer/chondrogenic layer (CL): contains chondrogenic cells
o **head of humerus does not have perichondrium**
● Cartilage cells – constitute only 3-5% of total cartilage mass
o Chondroblasts – synthesize cartilage matrix
▪ Location is just below chondrogenic layer of perichondrium

▪ Ovoid and positioned so that their longitudinal axes lie parallel to cartilage
surface (space occupied by chondroblast is called a lacuna – usually 1 cell per
lacuna)
o Chondrocytes – matured chondroblasts located in the lacunae and surrounded by matrix
▪ Divide by mitosis to form groups of 4-8 chondrocytes (referred to as Isogenous
groups of cell nest) – suggestive of internal growth (expansion) of cartilage
● As they mature the move apart by their own synthesis of ECM resulting
in growth of cartilage (essentially creating their own ECM)
● This is interstitial growth!
▪ Located deeper to the chondroblasts and are more spherical

▪ TEM – irregular surfaces, extensive Golgi apparatus, abundant rER, euchromatic

nucleus, and lipid droplets/glycogen deposits
● Extra Cellular Matrix (ECM) – constitutes >95% of cartilage volume and composed of amorphous
ground substance and collagen fibrils
o 15% collagen (mostly Type II – about 80%) with small percentage of Type III, VI, XII, XIV,
IX, and XI
o Not readily discernible in histological sections because fibrils are very fine and have
same refractive index as the ground substance (gives it “glassy” appearance and
therefore named “Hyaline” cartilage)
o Capsular Matrix – matrix adjacent to chondrocyte, poor in collagen but rich in GAGs
(stains deeply basophilic and with PAS)
o Territorial Matrix – matrix found around isogenous groups (surrounds the group of 4-8
chondrocytes)
o Interterritorial Matrix – matrix in-between isogenous groups
o Sulfated groups present in GAGs (hydrophilic) enable easy diffusion of nutrition to the
cell (also provides resilience)
o Proteoglycans provide immense strength to the matrix so that cartilage can function as a
model for bone formation (proteoglycan aggregate monomers)
● Hyaline Cartilage Growth: 2 types
o Appositional – occurs from chondrogenic cells in the perichondrium differentiating into
chondroblasts, forming a new layer of cartilage around the periphery of the existing
cartilage (increases the width or thickness of cartilage on top of pre-existing cartilage)
▪ Must have perichondrium in order to have appositional growth!
 o Interstitial – occurs in young cartilage from cell divisions within the cartilage (increases
the length from within the cartilage – isogenous groups)
● Hormonal influence – growth hormone, thyroxine, and testosterone increase growth,
hydrocortisone and estradiol decrease growth
● Regeneration – hyaline cartilage regenerates very poorly and often the perichondrium forms
scar tissue (due to being avascular)
● Calcification – in old age hyaline cartilage can get calcified (calcified cartilage = bone is coming)

Articular Cartilage – a type of Hyaline cartilage found at articular surfaces (does not have a
perichondrium) that is in contact with articular surface of bone on opposite side (remnant of original
fetal cartilage template of developing bone that persists throughout adult life)

● Regions: Superficial (tangential) layer, intermediate (transitional) layer, deep (radial) layer, and
calcified cartilage (zone)
● Functions: supportive, cushioning, shock absorber, growth plate, model for bone formation
o ECM correlates to function; collagen, sulfated groups
● Appositional growth not possible due to lack of perichondrium
● Osteoarthritis results from wearing/breaking down of articular cartilage that leads to bone
formation
o formation/ initial stages are painless due to lack of blood vessels, pain starts when you
have the “bone-on-bone” grinding (no pain from cartilage itself)

Elastic Cartilage – locations include’ pinna of the ear, external acoustic meatus, eustachian tube, and
epiglottis

● Histogenesis – mesenchymal cells; contains chondroblast and chondrocyte cells and ECM is Type
II collagen fibers and elastic fibers, GAGs, and glycoproteins
o Must use special stain to distinguish from Hyaline cartilage (stain will show the elastic)
● Growth – appositional & interstitial; degeneration – does not readily degenerate; calcification –
never calcifies
● Is often found with hyaline cartilage, possesses a perichondrium, grows by apposition and does
not degenerate as readily as hyaline cartilage
● Matrix is identical to that of hyaline cartilage, except it also contains a network of elastic fibers
that impart to it a yellowish color
● Located where support with flexibility is required

Fibrocartilage – locations include; intervertebral disc (annulus fibrosis), at the pubic symphysis,
medial/lateral menisci, attachment of ligament to bone, intersections of some tendons and ligaments,
and articular discs

● Histogenesis – mesenchymal cells; contains fibroblast and chondrocyte cells and ECM is
predominantly Type 1 collagen, also contains Type II collagen and GAGs (predominantly
chondroitin and dermatan sulfates)
● Growth – only interstitial (due to absence of perichondrium); does not occur alone but is found
in conjunction with hyaline cartilage and other fibrous tissues
● Function – support and tensile strength (found places with shear stress)
 ● Absence of perichondrium, fibers arranged perpendicular to direction of stress, isogenous
groups are in linear fashion (longitudinal columns) due to the compression of collagen fibers
● Fibers seen with H&E and trichrome stain

Summary – compare/contract the 3 types; Hyaline, Elastic, and Fibrocartilage

Lecture 4: Cells of the Bone

DLA

Cells of Bone Tissue: Osteogenic Cell > Osteoblast > Osteocyte > Osteoclast

● Osteogenic Cells differentiate into osteoblasts

● Osteoblasts secrete organic unmineralized matrix called osteoid and when entrapped in matrix
become osteocytes (maintain bone tissue)
● Osteoclasts are multinucleated giant cells responsible for resorbing and remodeling bone

Osteogenic (or Osteoprogenitor) Cells – derived from embryonic mesenchyme stem cells and are found
in the periosteum and endosteum or both mature/immature bone

● Spindle-shaped with pale elongated/ovoid nucleus and light stained cytoplasm

● Differentiate into osteoblasts

Osteoblast Cells – derived from osteogenic cells and morphology is dependent on activity

● Cuboidal osteoblasts indicate high secreting activity, Flattened cells indicate declining
secretions
● Cells are able to divide; during synthesis they have well developed rER, free ribosomes, and large
Golgi (reflects proteins (collagen type I) and proteoglycan synthesis)
● Matrix vesicles containing ALP/pyrophosphate are released by exocytosis and are key in
calcification of unmineralized matrix
 ● Osteiod – initial secretion which is uncalcified; has Ca2+ binding protein, multi-adhesive
glycoproteins, alkaline phosphatase, and proteoglycans
● Matrix section entraps the osteoblast in a lacunae with its cytoplasmic processes extending into
canaliculi
o Eventually will calcify to form calcified bone and the space surrounding the processes
will become the canaliculi (ceases secretion and becomes an osteocyte)
o KNOW CANALICULI WELL
● Some osteoblasts become bone lining while others (majority) undergo apoptosis

Osteocytes – mature bone cells, derived from osteoblasts (each osteocyte is housed in its own lacunae
which is the approximate shape of the cell)

● Smaller dimensions and fewer organelles reflect decreased synthesis; increased amounts of
condensed nuclear chromatin, reduced rERand smaller Golgi (compared to osteoblast)
● Maintain communication and exchange nutrients/metabolites with other osteocytes via gap
junctions between narrow cytoplasmic processes extending through the canaliculi (each cell may
have more than 50 processes)
● Able to modify their immediate environment by responding to mechanical forces applied
o Increased mechanical stress > stimulation of osteoprogenitor cells > bone formation
o Decreased mechanical stress > secretion of metalloproteinases > breakdown of bone
matrix in lacunae and canaliculi > osteocytic remodeling
● Remodeling plays an important role in regulation of Ca2+ and Phosphorus
o Osteocytes live for 10-30 years and decline due to apoptosis, necrosis, senescence, or
remodeling
● 3 types; osteocyte, formative osteocyte, and resorptive osteocyte (related to morphology)

Bone Lining Cells – line the surfaces of bone that are resting (not actively remodeling)

● Very little cytoplasm, few organelles located in perinuclear region, cytoplasmic processes which
contact with each other and with osteocyte processes, gap junctions present
● Endosteal – lining all internal spaces in bone tissue (Haversian canal)
● Periosteal – lining the external surface of bone

Osteoclasts – are multinucleated (up to 50 nuclei possible) giant cells responsible for resorbing and
remodeling bone

● Derived from the fusion of blood-derived mononuclear hemopoietic stem cells from the
granulocyte-macrophage lineage
● Distinct eosinophilic cytoplasm with abundant lysosomes (lysosome enzymes include; Cathepsin
K, matrix metalloproteinases, carbonic anhydrase II, various others)
● Howship’s Lacunae – a “bay” or depression on the bone surface during process of remodeling
where osteoclasts are found
● 3 distinct regions: ruffled border, clear zone, basal region
o Ruffled Border – finger-like evaginations along Howship’s lacunae (indicate sites of
active bone resorption)
 o Clear Zone – region of cytoplasm that surrounds the ruffled border (contains
microfilaments which anchor osteoclast to bony surface and help isolate osteoclastic
activity)
o Basal Region – houses the organelles and numerous nuclei of the cell
● Bone Resorption – lysosomal enzymes (from Golgi) and H+ ions are released into confined space
between bone matrix and osteoclast’s peripheral clear zone, acidic environment facilitates the
dissolution of calcium phosphate from bone
o Decalcified bone matrix is broken down by acid hydrolases and collagenous/proteolytic
enzymes and cell resorbs the organic/inorganic material

Lecture

Bone Overview – the organs of the skeletal system

● Functions – protection of vital organs, attachment for skeletal muscle and locomotion,
maintenance of body form, calcium homeostasis (bone is main storage of Ca2+ in body), and
marrow reservoir for hematopoietic stem cells
● Classification – gross morphology; long, short, flat, irregular
● Composition – most bones are composed of bone (osseous) tissue; spongy and compact and
supportive connective tissue; periosteum and endosteum (located in every cavity that you will
find bone)

Bone Tissue – connective tissue specialized (matrix is different from any other tissue) for tensile strength

● Cells – osteocytes, osteoblasts, osteoclasts, bone lining cells, endosteal/periosteal cells

● Organic ECM – fibers (fibrous proteins) mainly Type 1 collagen (also contains types III, V, IX, and
X) and ground substance (proteoglycans, bone specific proteins, growth factors, and cytokines)
o Type X collagen is especially important for bone formation
● Inorganic ECM – calcium phosphate (in form of hydroxyapatite which is 50% of bone dry weight)
and other inorganics such as Mg2+, bicarbonate, citrate, and carbonates
● Stains – H&E of decalcified mature bone (special stain used to view, decalcified first for easier
way to view organic composition)
o Ground section of bone with India ink is best to observe compact bone (organic
material is removed)
● Immature (or woven) – found in developing fetus, remodeling sites, and aveolar sockets of
adults, non-lamellar, increased cellularity, more ground substance and randomly arranged
collagen fibers, remodeled and replaced by mature bone
● Mature (or lamellar) – forms the majority of the adult skeleton, type of tissue has a regular
arrangement of collagen and cells within the calcified matrix, organized as lamellae, very
resilient (fewer cells than immature and more organization and calcium)

Mature Bone – bones are composed of 2 types of mature osseous tissue; compact and spongy bone
which mainly differ in how the bone mineral is organized

● Compact – appears solid of solidified ECM, forms the outer part (cortex of bones), very few
spaces (if you see OSTEON then think compact bone)
 o Arranged in cylindrical osteons or Haversian systems (central canal lined by endosteal
cell)
▪ Haversian systems are NOT all uniform and in parallel array (due to constant
bone remodeling)
o Osteons – osteocytes in their lacunae
o Canaliculi – body channels that allow for osteocyte processes to communicate with
neighboring cells and Haversian Canal (contain osteocyte processes)
o Each lamella consists of multiple parallel arrays of collagen fibers
▪ Adjacent lamellae have alternating directions of collagen fibers providing great
strength despite light weight
o Concentric lamellae of matrix with cells on the edges around a central vascular channel
(Haversian canal) through which blood vessels, lymph, and nerves can travel
o Base of individual osteons are Volkmann’s Canals which allow blood vessels, lymph
vessels, and nerve to travel across compact bone linking neurovasculature of Haversian
canals to the periosteum/endosteal marrow cavity
o Haversian Canals and Volkmann’s Canals run perpendicular to each other!
o Types of Lamellae: each separate lamellae has its collagen fibers run in the same
direction
▪ Concentric – form the osteon, 4 – 20 layers around Haversian Canal

▪ Inner Circumferential – concentric rings arranged next to endosteum

▪ Outer Circumferential – concentric rings deep to the periosteum

▪ Interstitial – remnants of old remodeled osteons between newly laid down

osteons
● Spongy (Cancellous) – web or sponge-like arrangement, forms most of bulk of bones, most of its
volume occupied by marrow
o Filled with spaces that are interconnected by bony trabeculae (thus = trabecular bone)
o Red (hemopoietic cells) or yellow (primarily fat) bone marrow
o Do NOT have osteons
o Within a single trabecular there are concentric lamellae with osteocytes in lacunae
connected to one another via canaliculi, similar to tissue arrangement in osteons of
compact bone
o Do NOT have central canals or perforating canals containing blood vessels, lymph, or
nerves
▪ Vessels/nerve travel through spaces between trabeculae and do not need
separate passageways
● Endosteum – is the non-calcified vascular connective tissue layer covering internal surfaces of
bone (ie. Marrow cavities, inner lining of cortex, Haversian and Volkmann's canals)
o Single layer of endosteal cells, osteoprogenitor cells differentiate into osteoblasts, bone
lining cells
 ● Periosteum – is the non-calcified vascular connective tissue layer covering external surfaces of
bone, except over articular surfaces (extremely sensitive layer)
o 2 layers: anchored to outer circumferential and interstitial lamellae by perforating or
Sharpey’s Fibers
▪ Outer Layer - made of mostly dense collagenous connective tissue containing
blood vessels
▪ Inner layer – contains osteoprogenitor cells and bone lining periosteal cells

Bone Formation – 2 model of bone development described; immature bone forms first in both
processes and is later replaced by mature bone and remodeling continues throughout life (slower than
mature bone)

● Intramembranous bone formation involves development within layer of condensed

mesenchyme (Examples of bone formed in this fashion include most of the flat bones of the
skull)
o Pluripotent mesenchymal cells condense to form a primary ossification center, from
which osteoblasts differentiate and begin secreting unmineralized matrix or Osteoid
o Calcification; osteoblasts become trapped in their own matrix to form osteocytes
o Osteoblasts line outside of spicules (spicules = tiny pieces of bone)
o Ossification centers expand to form bony spicules as more osteoblasts aggregate and
begin secreting
o Fused spicules form bone trabeculae which eventually forms primary (immature) bone
o As spicules continue forming, surrounding mesenchyme condense to form connective
tissue which later forms periosteum
o Blood vessels invade area at same time that undifferentiated mesenchymal cells give rise
to bone marrow cells
o Monocytes come forming osteoclasts and remodeling of primary bone results in mature
compact and spongy bone tissue

● Endochondral bone formation occurs via a substitution of bone tissue on a cartilage model
o Outline of bone in hyaline cartilage
o Formation of periosteum and subperiosteum bone collar for support in diaphyseal
region (cartilage cells are hypertrophic)
o Cartilage matrix is calcified
o Blood vessels erode into calcified cartilage
o Formation of periosteal bud consisting of osteogenic cells and blood vessels (primary
ossification center)
o The subperiosteal collar (diaphyseal bone) becomes thicker and bone forms on the
calcified cartilage complexes
o Secondary ossification centers (in a similar manner) form in the epiphyses
o Epiphyseal cartilage (epiphyseal growth plate) is formed between the primary and
secondary ossification centers
o Epiphyseal plate disappearance occurs at different times
 o Fusion of diaphyseal and epiphyseal marrow cavities

Epiphyseal Plate – continues to grow by adding new cartilage at the epiphyseal end while it is being
replaced by bone at the diaphyseal end (diaphyseal and epiphyseal bone become continuous (connect)
around age 20)

● Contains 5 histologically distinct zones:

o Zone of reserve cartilage (ZRC): cartilage with small, randomly arranged inactive
chondrocytes
o Zone of cell proliferation: rapid mitotic divisions give rise to rows of cartilage cells
o Zone of hypertrophy: the chondrocytes are greatly enlarged and the cartilage matrix
between neighboring cells becomes thin
o Zone of calcified cartilage: lacunae coalesce and the inter-lacunar matrices become
calcified, causing apoptosis of chondrocytes
o Zone of resorption: bone is beginning to be elaborated upon the calcified cartilage, and
osteolytic activity begins to resorb the calcified bone-cartilage complex

Development of Osteons: step-wise fashion outside the periosteum layer

● Longitudinal ridges form along the bone and osteogenic cells in the periosteum transform into
osteoblasts
● Osteoblast start producing bone matrix which form ridges that close off periosteal capillaries as
they meet
● Periosteum lining the newly formed canal becomes the endosteum and starts forming concentric
lamellae to form the osteon

Bone Growth and Remodeling – elongation of bones is a result of interstitial growth of cartilage at
epiphyseal growth plate (interstitial growth does NOT occur in bone)

● Bone increases in width by appositional growth from periosteum (osteoprogenitor cells in the
periosteum)
● Remodeling of bone is a coordinated action between osteoblasts and osteoclasts

Bone Repair – fractured bone results in damage to bone matrix, bone cells in the region, as well as blood
vessels supplying the area

● Hemorrhaging is followed by blood clotting, and macrophages remove much of the debris via
phagocytosis
● Fibroblasts proliferate in the periosteum and endosteum and surround the area internally and
externally to isolate it
● Chondrocytes also differentiate from this connective tissue and elaborate cartilage
● A fibrocartilaginous callus is formed both internally and externally (soft callus)
● Simultaneously osteoblasts differentiate from the periosteum and endosteum which begin to lay
down bony matrix on the outer surface of the callous (intramembranous ossification)
● Cartilage is replaced by bone via endochondral bone formation as an osteogenic bud penetrates
callus and matrix is laid down within the callus (Endochondral ossification), this forms a hard
callus of primary (immature) bone
 ● Bony callus is eventually resorbed and replaced with secondary bone as the repair process
continues

Bone Nutrition – nutrition affects bone development thus the need for adequate nutrition (especially in
children)

● Nutrients from Diet:

o Diets low in protein result in deficiency of amino acids essential for collagen synthesis by
osteoblasts
o Lack of calcium, either from low intake or inadequate absorption by the small intestine
(due to a lack of vitamin D), results in poorly calcified bone, which leads to rickets in
children and osteomalacia in adults
o Vitamin D is necessary for proper ossification (excess may actually cause bone
resorption)
o Vitamin A deficiency inhibits proper bone formation and growth (excess accelerates the
ossification of the epiphyseal plates and makes bone fragile leading to smaller stature)
o Vitamin C is necessary for collagen formation - deficiency results in scurvy, characterized
by poor wound healing and bone repair after fractures
● Hormones: exert an influence on bone formation by regulating availability of tissue components
as well as the process of remodeling and formation
o Parathyroid Hormone (PTH) – indirectly stimulates osteoclasts to resorb and release
calcium, thus elevating blood calcium levels, may activate osteocytes to initiate
osteolysis
▪ In excess renders bone more susceptible to fracture and subsequent deposition
of calcium in arterial walls and certain organs such as the kidney
o Calcitonin – inhibits matrix resorption and thus prevents the release of calcium
o Sex Hormones – affects bone growth by influencing epiphyseal ossification, excess =
small stature develops; deficiency = tall stature results
o Pituitary Growth Hormone (GH) – stimulates epiphyseal cartilage growth, so that an
excess in childhood produces a giant, while a lack produces a dwarf
▪ In adulthood, excess GH results in acromegaly

Osteoporosis – means “porous” bone and results from progressive loss of bone density leading to
increased risk of fracture

● Primary Type 1 - Post-menopausal women estrogen levels decrease with menopause (estrogen
normally limits activity of osteoclasts)
o 33% of postmenopausal women affected – hormone therapy, selective estrogen
receptor modulator therapy
● Primary Type 2 – elderly people in their 70s-80s
o Secondary osteoporosis develops as a consequence of drug therapy (corticosteroids) or
other disease process (malnutrition, weightlessness, metastatic cancer)