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CYTOLOGY

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Title: CYTOLOGY


1
CYTOLOGY NUCLEUS
2
Light Microscopy vs. Electron Microscopy
  • Light Microscopy see cells, not organelles,
    usually colored by stain preparation (HE most
    often)
  • Electron Microscopy see organelles and small
    details, black and white color, resolving power
    down to 1nm
  • STAINS (on Med Micro Atlas)
  • HE Periodic Acid-Schiff Silver
    Impregnation
  • Verhoeffs w/ hemotoxylin Methylene blue
    eosin

3
  • Eukaryotes HAVE NUCLEUS
  • Prokaryotes DONT HAVE PROPER NUCLEUS
  • Karyotype
  • Chromosomes visible late prophase line up late
    metaphase (mitotic figure)
  • Chromatin (dsDNA)?Wrapped around Histones to form
    nucleosomes (beads on a string)?Solenoids?Chromoso
    me loops and Superhelices?Condensed Chromosome
  • 23 pairs, 46 total chromosomes at G0 (22
    autosomal pairs 1 sex chr. pair)
  • In females, XX. One X inactivated to form Barr
    body located on periphery of nucleus adjacent to
    nuclear membrane.
  • Metacentric Chromosomes
  • Submetacentric Chromosomes
  • Acrocentric Chromosomes
  • Telocentric Chromosomes
  • Telomerasepreserves the lagging strand during
    replication (S phase) in germ cells and cancer
    cells

p arm
Centromere
q arm
4
Nuclei vs. Cytoplasm
  • Cytoplasm
  • Stained by Eosin
  • Nuclei
  • Stained by hemotoxylin
  • Darker areasheterochromatin
  • Lighter areaseuchromatin active replication and
    txn

5
  • Hematoxylin ACIDOPHILIC (loves acids) stain
  • Blue/Purple Color
  • Nucleus (Deoxyribonucleic acids)
  • Prominent nucleolus in protein-secreting cells
  • RNA/ribosomes (Ribonucleic acids)
  • More blue color in cytoplasm of protein-secreting
    cells
  • EosinBASOPHILIC (loves bases/non-acids) stains
  • Pink Color
  • Stains everything else pretty much by default
    more prominent in cytoplasm

6
PANCREAS
Basophilic portion of cytoplasm in secreting
cell (RER)
Nuclei
Eosinophilic portion of Secreting cell
7
Nucleus
  • Surrounded by double-folded nuclear membrane with
    nuclear pores for transport (outer nuc. membrane
    continuous w/ RER membrane in cytoplasm)
  • During mitosis, nuclear lamins are phosphorylated
    to break apart the membrane and later
    dephophorylated to reform
  • Nucleolusprominent in protein secreting cells
    (also lots of RER in cytoplasm)
  • Heterochromatindark staining at center and
    periphery of nucleus txnally inactive indicates
    loc. of nuclear pores
  • Euchromatinlighter staining portion of nucleus
    txnally active

8
Nucleolus
  • Prominent in protein-producing cells give
    nucleus a bulls eye appearance
  • Associated in ribosome assembly
  • Associated with the stalks of acrocentric
    chromosomes (also called nucleolar organizer
    region)

9
  • FREE RIBOSOMES IN CYTOPLASM
  • Synthesize protein that will be used within the
    cell
  • ROUGH ENDOPLASMIC RETICULUM
  • Makes protein that will be secreted or
    incorporated into plasma membrane
  • On EM, prickly looking
  • SMOOTH ENDOPLASMIC RETICULUM
  • Continuous with RER, but smooth (no ribosomes)
  • Involved in lipid/steroid production (seen in
    cells with mitochondria with tubular cristae)
  • Regulates calcium homeostasis
  • Involved in detox of alcohol toxins
  • Involved in glycogen metabolism

10
GOLGI APPARATUS
  • Comprised of stacked cisternae
  • Processes protein from RER before its sent to
    membrane
  • Protein enters transfer vesicles to immature
    (cis) face and exits mature (trans) face in
    secretory vesicles bound towards the plasma
    membrane for integration into membrane or
    secretion
  • Source of lysosomes

11
Inclusions
  • Glycogen granules
  • Look like dark, small, irreg-shaped, non-membrane
    bound grape clusters
  • Lipid droplets
  • Clear, round, non-membrane bound
  • Secretory vesicles
  • MEMBRANE-BOUND
  • Constitutive releasefew vesicles in cytoplasm
  • Regulated releasemany vesicles in cytoplasm
  • Lysosomesmembrane-bound, containing acid
    hydrolases
  • Primarycontains hydrolytic enzymes, dark and
    regular look
  • Secondaryphagolysosome, junky cytoplasm
  • Tertiaryindistinguishable from secondary,
    residual bodies, lipofuscin granules

12
  • PEROXISOMES
  • From SER pre-existing peroxisomes, contain
    oxidative enzymes (catalase), involved in fatty
    acid oxidation
  • (Wont have to identify on EM)
  • PROTEOSOMES
  • core regulatory particle
  • Degrade intracellular protein targeted for
    destruction with ubiquitin

13
Mitochondria
  • Inner membrane (site of aerobic resp
    foldscristae) outer membranes
  • Self-replicating
  • May have small, dark, round, CA2 deposits in
    matrix in addition to mitochondrial DNA enzymes
  • TUBULAR cristaecharacteristic of
    steroid-secreting cells, (context cluesabundant
    SER), look at picture on blackboard EM

14
MICROTUBULES
  • alpha-tubulin beta-tubulin alpha/beta
    heterodimer "tubulin. 13 tubulin filaments in
    a spiralone microtubule.
  • Functions cell division, organelle mvmt,
    cytoskeleton, cilia, flagella, centrioles
  • Axoneme of flagella cilia 9x22 microtubule
    arrangement
  • Centriole and basal body (proximal Axoneme)
    9x30

15
MICROFILAMENTS/ACTIN
  • Cytokinesis (cytoplasm division)
  • Anchoring of actin to form terminal web in zonula
    adherens
  • Mvmt of motile cells organelles
  • Microvilli anchor to terminal web
  • Muscle contraction
  • Inflammatory roleneutrophil recruitment

16
INTERMEDIATE FILAMENTS
  • Cytoskeletal element 2 non-polar coiled elements
    (versus polar microtubules)
  • Nuclear Lamins
  • Source of neurofibrillary tangles in Alzheimers

Intermediate Filament Associated cell type
Cytokeratin Epithelial Cells
Vimentin Mesenchymally-derived cells
Desmin Muscle cells
Glial Fibrillary Acidic Protein Glial Cells in CNS
Neurofilaments Neurons in CNS and PNS
17
Zonula Occludins
  • TIGHT JUNCTIONS
  • Goalform impermeable barrier to fluids, found in
    multiples
  • Typically located towards apical (lumenal)
    surface of cell, on lateral side
  • Join together the cytoskeletons of adjacent cells
  • Connect to ACTIN, or Microfilaments

18
Zonula adherens
  • More basally located than tight junctions (z.
    occludens)
  • Similar to fascia adherens
  • Appear as bands encircling the cell, forming
    actin contractile ring
  • Cell-cell junctions linked to ACTIN in the
    cytoskeleton
  • cadherin alpha-catenin beta-catenin actin

19
Macula adherens
  • DESMOSOMES, spot junctions on lateral sides of
    cell
  • Link to cytokeratin or desmin filaments
  • INTERMEDIATE FILAMENTS
  • Random arrangement on cell
  • Allow cell to resist shearing stress
  • Found in simple and stratified squamous epithelium

20
Lumen
Apical
ZONULA OCCLUDINS
ZONULA ADHERENS
MACULA ADHERENS
Basal
21
Gap Junctions
  • Transmembrane hemi-pore
  • Allow transfer of ions
  • Synchonized contraction and function among
    adjacent cells

Cell adhesion molecules
  • Selectins
  • Integrins
  • Immunoglobulins
  • Cadherins

22
MODIFICATION OF CELL SURFACE
  • BASAL INFOLDINGS increase surface area on basal
    (basement) side of cell increases the surface
    area available for transportation, therefore they
    are frequently associated with mitochondria
  • MICROVILLI increase the apical (lumenal)
    surface area often have a fuzzy appearance

EM View
LM View
Epithelium with Microvilli, (Carbohydrate, NOT
same as cilia)
23
Which of the following is not an integral
membrane protein complex?
  • A. Cadherin
  • B. Immunoglobulin E
  • C. Integrin
  • D. Selectin
  • E. Cytoskeleton

Which of the following is true of the smooth ER?
  • A. Special name for Golgi apparatus in striated
    muscle
  • B. Can be a secondary site for protein synthesis
  • C. Involved in calcium homeostasis
  • D. Source of lysosomes

24
Which of the following is NOT an integral
membrane protein complex?
  • A. Cadherin
  • B. Immunoglobulin E
  • C. Integrin
  • D. Selectin
  • E. Cytoskeleton

Which of the following is true of the smooth ER?
  • A. Special name for Golgi apparatus in striated
    muscle
  • B. Can be a secondary site for protein synthesis
  • C. Involved in calcium homeostasis
  • D. Source of lysosomes

25
Last words of wisdom
  • When in doubt, its probably a
  • MITOTIC FIGURE

Source http//education.vetmed.vt.edu/Curriculum/
VM8054/Labs/Lab19/IMAGES/MITOTIC20FIGURES20GUT2
0SMALL.jpg
26
EPITHELIUM
27
  • Simple squamous epithelium
  • -lining epithelium of an artery/vein
  • -lines alveoli of lungs
  •  
  • Endothelium
  • -simple squamous epithelium lining the lumen
    of blood vessels
  •  
  • Mesothelium
  • -simple squamous epithelium that covers/lines
    body cavities

Simple Squamous epithelium
Alveoli of lung
28
Simple Squamous epithelium Kidney
Simple Cuboidal Epithelium
Thyroid
http//www.cytochemistry.net/microanatomy/Endocrin
es/thyroid1.jpg
29
Simple Columnar Epithelium Jejunum
Stratified Squamous Epithelium Lip
  • Keratinized, stratified squamous epithelium of
    skin
  • -stains positive for cytokeratin intermediate
    filaments
  • Has striated border made of microvili

30
Stratified Cuboidal Epithelium Skin
  • Secretory portion

Stratified Columnar Epithelium Tongue
  • Excretory duct of a sweat gland is located within
    dense CT of the dermis

31
Psuedostratified columnar epithelium Trachea
Transitional Epithelium Bladder
Cilia
  • Distensible
  • Nuclei are at different levels, however
  • all cells contact basal lamina
  • Aka- respiratory epithelium

32
Tonofilaments -intermediate filaments in
epithelial cells -attached directly to
desmosomes/hemidesmosomes -Macula adherens (
think MAD Tony) Terminal Web -comprised of
actin filaments -actin filaments of terminal web
attach to Zonula Adherens   Gap Junctions -used
for communication, ions move between adjacent
cells Axoneme -9x22 arrangement of
microtubules in a cilium or flagellum
33
  • The lining epithelium of an artery is classified
    as
  • Simple squamous epithelium
  • Stratified squamous epithelium
  • Simple cuboidal epithelium
  • Stratified cuboidal epithelium
  • Transitional epithelium

34
  • In epidermal cells, tonofilaments are attached
    directly to the
  • Basal lamina
  • Desmosomes
  • Nuclear envelope
  • Zonula adherens
  • Zonula occludens

35
  • The terminal web of an absorptive epithelial cell
    of the gastrointestinal tract is comprised of
  • Actin filaments
  • Cytokeratin filaments
  • Desmin
  • Fibronectin
  • Microtubules

36
Buzz Words Tonofilaments -intermediate filaments
in epithelial cells -attached directly to
desmosomes/hemidesmosomes -Macula
adherens   Cadherens -present in extracellular
space, facilitate cell anchoring   Keratinized,
stratified squamous epithelium of skin -stains
positive for cytokeratin intermediate
filament   Simple squamous epithelium -lining
epithelium of an artery/vein -lines alveoli of
lungs   Endothelium -simple squamous epithelium
lining the lumen of blood vessels   Mesothelium -
simple squamous epithelium that covers/lines body
cavities  
37
Endothelial and Mesothelial cells -differ from
all other epithelial cells because theyre
derived from mesoderm-gt mesenchymal cells-gt
multiocular/unilocular adipocytes, purkinje
fibers of heart -vimentin -(all other epith
cells are from ectoderm or endoderm)   Simple
Columnar -intestines   Stratified cuboidal
epithelium -excretory duct of a sweat gland,
located within dense CT of the dermis   Pseudostra
tified epithelium/respiratory epithelium -nuclei
are at different levels, however all cells
contact basal lamina -lines trachea   Transitiona
l epithelium -urinary tract -distensible   Mitoc
hondria -found within basal infoldings
characteristic of epithelial lining cells of GI
tract  
38
Axoneme -9x22 arrangement of microtubules in a
cilium or flagellum    Terminal Web -comprised
of actin filaments -actin filaments of terminal
web attach to Zonula Adherens   Glycocalyx -glyco
proteins, absorption   Reticular fibers- highly
glycosylated   Holocrine secretion -whole cell
dies and product released at time of cell
death -sebaceous glands   Merocrine
secretion -product released from cell and only
product leaves -sweat glands (excretory
ductstratified cuboidal epith), mast
cell   Apocrine -product and a little cytoplasm
leaves  
39
Constitutive secretion -continuous, rapid
secretion, product not stored in
cell -fibroblast   Regulated secretion -continuo
us, rapid secretion -products not stored in
cell   Exocrine duct -secretory portion makes
product -duct travels through to surface -hair
follicle with attached sebaceous
gland   Endocrine duct -no duct, no longer
attached to any body surface -product released
into vasculature -anterior lobe of pituitary
gland if you can see a duct draining to the
surface you know its exocrine. However, not all
pics will show you the duct at the surface, so
you need to know location, location, location to
determine if exocrine or endocrine gland!  
40
Gap Junctions -used for communication, ions move
between adjacent cells -wave of contraction
spread cell to cell in layers of smooth muscle by
ionic flow through gap junctions -synchrounous
spread of a wave of contraction through
cardiocytes forming the atrial myocardium, uses
gap junctions
41
MUSCLE CONNECTIVETISSUE
42
Fibroblasts and ECM
Proteoglycans Glycosaminoglycans Protein
Core Glycosaminoglycans Polysaccharide made of
glycosamine (disaccharide subunit). GAGs have a
negative charge and thus attract WATER GEL
ECM SECRETED BY FIBROBLASTS
43
(No Transcript)
44
EOSINOPHIL
LYMPHOCYTE
FIBROBLAST
MACROPHAGE
PLASMA CELL
45
Myoepithelial cell WHERE IS IT???
46
EMs
47
What to think about Intercalated
disks Striations Nucleus location Branching
Maculae adherens (desmosomes) Fascia
adherens Gap junctions ion
48
Purkinje Fibers
Part of myocardium (subendocardium) NOT
endocardium Conduction system of the heart but
not part of the CNS Lots of gap junctions and
glycogen granules
49
Question 1
  • Identify the cell
  • Fibroblast
  • Fibrocyte
  • Macrophage
  • Lymphocyte
  • Plasma Cell

50
Question 1
  • Identify the cell
  • Fibroblast
  • Fibrocyte
  • Macrophage
  • Lymphocyte
  • Plasma Cell

51
Question 2
  • Which of the following images include cells that
    are contractile?
  1. A
  2. B
  3. C
  4. D
  5. A-C
  6. A-D

B
A
D
C
52
Question 2
  • Which of the following images include cells that
    are contractile?
  1. A
  2. B
  3. C
  4. D
  5. A-C
  6. A-D

B
A
D
C
53
NERVOUS TISSUE MITOSIS
54
NEURO
  • Organization
  • CNS Brain and SC
  • PNS Afferent/Efferent
  • Neural Crests Branch away from neural tube to
    become a variety of Sxs
  • Ectoderm?Neuroectoderm?Neuroblast?Neurons and
    Glia
  • Cell
  • Body, aka perikaryon, aka soma RER ? nissl
    substance and NTs!
  • Axons and Dendrites Information gathering and
    transmission

55
  • Neuro, Contd
  • Dendrites lots of MTs to direct axonoplasmic
    flow
  • Axons Origin of APs! Hillock Ion chanels!
  • Non-nissl origin
  • Three Flow Types FF, SF and REV
  • Neurofilaments, MTs and organelles
  • Synapse Axon Terminal target (dendrite)
  • Exocytosis of NTs
  • MEP, aka, NMJ 1 motor nerve and all the muscle
    fibers it innervates
  • Nerve Fx
  • Membrane depolarized to threshold? signal
    travels down axon (faster in myelinated cells)?
    AP reaches terminal?Cainflux?exocytosis?reuptak
    e/breakdown of NTs

56
Nuclues and Nucleolous Fx? NeuropilGlial
cells Type?Nissl Sub
57
  • Neuro, contd
  • Cell Types Location, Location, Location!
  • Multi Polar many dens, one axon
  • Unipolar Peripheral Ganglions
  • Bipolar Special Senses

58
  • Neuro, contd
  • Stroma v. Parenchyma
  • Glial Cells
  • - CNS Astrocytes Big one, make GFAP
  • Oligodendrocytes Myelin in CNS
  • Microglia Phagocytes on CNS
  • Epyndyma lines ventricles and produces CSF
  • PNS Schwan Cells Neural Crest derivatives
  • Mesaxon, Nodes of Ranvier and Incisures of
    Schmidt-Lanterman(Osmuim Tetroxide)
  • Injury
  • Wallerian Degeneration nerve process injury
    where axon looses contact/comm w/
  • cell body. Regeneration can occur if Schwann
    Cells are maintained and can form a hollow tube
    Neurofilaments will grow through tube

59
Schwann CellNucleus/FibrocyteNodes of Ranvier
and ISL
60
Cross SectionAxons and Schwann cellsEpineurium,
perineurium and Endoneurium
61
Who/Where am I?
62
Who/Where am I?
  • Cerebellum? Purkinje Cells. Multiple dendrites
    extending into molecular layer, while axons
    extend into white matter traveling out of the CNS
    (efferents)

63
Who/Where am I?
64
Who/Where am I?
  • Pyramidal cells in cerebral cortex. Dendrites
    extending out the apex and axons descending from
    the base

65
Meiosis and Mitosis
66
Meiosis and Mitosis
  • The prophase, metaphase, anaphase and telophase
    portions of the cycle are essentially similar for
    Mieosis 1 (Reduction) and Mitosis
  • For purposes in this class, understand the steps
    at each phase and that Meiosis II
    (Condensation)is the separation of chromosomes to
    individual chromatids.

67
Question 1
  • Which of the following statements is correct?
  • Action potentials travel down the axon in the
    antereograde direction, reaches the axon terminal
    and after the influx of calcium, release NTs via
    endocytosis
  • Cells with a high amount of nissl substance
    produce large amounts of steroids
  • Incisures of Schmidt-Lanterman represent channels
    of cytoplasm between flattened Schwann membrane
    that allows cell communication
  • It is easy to discern a bipolar neuron from the
    cribiform plate with HE stain by the shape of
    the nucleus under medium power stain.

68
Question 1
  • Which of the following statements is correct?
  • Action potentials travel down the axon in the
    antereograde direction, reaches the axon terminal
    and after the influx of calcium, release NTs via
    endocytosis
  • Cells with a high amount of nissl substance
    produce large amounts of steroids
  • Incisures of Schmidt-Lanterman represent channels
    of cytoplasm between flattened Schwann membrane
    that allows cell communication
  • It is easy to discern a bipolar neuron from the
    cribiform plate with HE stain by the shape of
    the nucleus under medium power stain.

69
Question 2
  • Which of the following statements is incorrect
  • The gray matter of the spinal cord is composed of
    cell bodies while the white matter is composed of
    myelinated axons
  • The neuropil seen under a microscope is made up
    of everything but the cell bodies, including the
    axons, dendrites, glial cells and all of their
    synapses
  • Nerves cut in cross section can be best be
    discriminated from smooth muscle by the presence
    on a dense perineurium surrounding the bundle
  • Myelin, on HE stain is weakly stained because it
    is made up almost entirely of phospholipid that
    does not accept stain very well.

70
Question 2
  • Which of the following statements is incorrect
  • The gray matter of the spinal cord is composed of
    cell bodies while the white matter is composed of
    myelinated axons
  • The neuropil seen under a microscope is made up
    of everything but the cell bodies, including the
    axons, dendrites, glial cells and all of their
    synapses
  • Nerves cut in cross section can be best be
    discriminated from smooth muscle by the presence
    on a dense perineurium surrounding the bundle
  • Myelin, on HE stain is weakly stained because it
    is made up almost entirely of phospholipid that
    does not accept stain very well.
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