Histology lecture: Cytoplasm 4

The Cytoskeleton  The assembly of G-actin monomers through

the help of complex polypeptide, forms the F-
actin filament
 The thinnest among the three filaments of the
cytoskeleton
 Also called actin filaments because they are
composed of actin
 Are complex array of protein polymers that  One important use of microfilament is in
determine shapes of cells, play an important muscle contraction, during muscle contract a
role in the movements of organelles and protein called myosin-bind to actin and it said
cytoplasmic vesicles as well as in the to walk along it does it create the contractile
movement of entire cell motion
 Which includes:  Microfilaments are polarized, they use the
 Microtubules negative end to bond to cell structure and grow
 Microfilaments/Actin filaments from the positive end
 Intermediate Filaments o Grow means G-actin monomer is
 Network of protein fiber that permeate all of being added to the positive end
the cell through ___ and other complex
 Just like the bones of the body, it gives the polypeptides
structure of the body, shape, and ability to  As they grow they pushed against the cell to
resist pressure create tension, giving the cell the stability,
 Also has the ability to move molecules or the shape, and structure
product within the cell, has the ability to  Microfilaments are concentrated beneath the
organize the organelles and the structures cell membrane and in cell extensions like
found within the cell microvilli
General Function of the Cytoskeleton
Structural Subunit G-actin monomers (Globular actin)
 provides structural support to the cell Overall Two intertwined filaments of f-actin
structure
Movement diameter 5-7 nm
 Assists with cytosol streaming and cell Monomeric Globular g-actin (42 kDa)
motility proteins
Polarity + and – ends
 Helps move organelles and materials Relative Dynamic
throughout cell stability
General Concentrated beneath cell
 Helps move chromosomes during cell location
division membrane; in cell extensions like
microvilli
Key Contract and move cells; change
functions
cell shape; cytokinesis; cytoplasmic
transport and streaming

Intermediate Filaments
3 Types of Filament
Microfilament

 Made up of proteins, more than a dozen that

serves as subunits of various intermediate
filaments, this subunits are coiled

Subunit Antiparallel tetramers of two rod-

Histology lecture: Cytoplasm 4

like dimer  vimentin

Overall Cable of four intertwined o desmin
structure
protofibrils, each consisting of o GFAP (Glial fibrillary acidic protein)
bundled tetramers associated end-to-
 Neurofilament
end
diameter  Lamins
8-10 nm
Monomeric Various – a-helical rol-like proteins  The presence of a specific type of intermediate
proteins filament in certain tumors can often reveal the
(-55 kDa)
Polarity No apparent polarity cell of origin of the malignancy or tumor. This
Relative stable can help in treating the malignancy or tumor.
stability
General
An this can be identify through the use of
Arrayed throughout cytoplasm; at
location immunocytochemical methods, like GFAP, it
desmosomes; inside nuclear envelop
Key is use to identify astrocytoma (most common
Strengthen cell and tissues structure;
functions
maintain cell shape; maintain type of brain tumor)
nuclear shape (lamins)
Microtubules
Major Classes and Representatives of IFP, their
sizes and locations.
More than a dozen proteins serves as subunits of
various intermediate filaments, these subunits can
be localized by immunohistochemistry in different  Thickest of the three fibers
types of cells. Some of which are used to help in  Rigid hollow tube made up of alpha and beta
the diagnosis of certain diseases that results from tubulin, together they create helical structure
their disruption that wined up creating a hollow center
Class Protein Size Cell Disease  Th positive end is where the microtubules
(kDa) Distributio Involvement (If
n Known)
grow, the negative end is found inside the cell,
I Acidic 40-65 Epithelial Certain skin- usually found in a region given by the MTOC
cytokerati Cells blistering (microtubule-organizing center) known
n disorders
II Basic 51-68 Epithelial Myopathies as centrosomes. It’s the centrosome
cytokerati cells region that contains our centriole
n
III Desmin 53 Muscle  Essentially, our microtubules are made with in
cells our centrosome. If centrosome is responsible
Synemin 190 Muscle Alexander
cells disease for cell division, this implies that the
GFAP 50 Astrocytes microtubules are also involve in the cell
(less in
other glial division
cells)  Microtubules separates chromosomes during
Peripherin 57 Neurons
Vimentin 54 Mesenchy cell division and create mitotic spindle fibers.
mal cells  They transports things within the cell just like
IV NF-L 68 Neurons
NF-M 110 Neuron
highway, it transport along the microtubule
NF-H 130 Neurons under the control of motor proteins which use
a- 55 Embryonic also energy called ATP in moving larger
internexin neurons
V Lamins 62-72 Nuclei of Cardiomyopath structures. Example the kinesin, it carries
all cells y; muscular material away the MTOC near the nucleus
dystrophies;
progeria towards the positive end of the microtubules,
VI Nestin 230 Neural also known as the anterograde transport. The
stem cells
cytoplasmic dynein carries materials along
microtubules in the opposite direction
Intermediate filament proteins with particular
biological, histological, or pathological importance  It is also important in the movement of the cell
includes: itself. It is also responsible for formation if
structures that lets the cells moves like flagella
 keratin/cytokeratin
Histology lecture: Cytoplasm 4

and cilia, in prokaryotes. In eukaryotes, it use  Microtubules has special dynamic properties,
different in a population of a microtubule at any point in
Subunit Heterodimer of aB-tubulin time, a subset of microtubules are rapidly
Overall Hollow tube with a wall of 13 growing while others are quickly shrinking.
structure
parallel protofilaments Although sometimes, some microtubules sits
diameter 25 nm in a pause state.
Monomeric a and B tubulin (54 kDa)
proteins  Each microtubule, which randomly growing
Polarity + and – ends and shrinking states, sometimes changing back
Relative Dynamic in cytoplasm; stable on and forth several times in the course of their
stability
axonemes lifetime. The combination of growth,
General Radiating through cytoplasm from shrinkage, and rapid transition between the
location
concentration at centrosomes; two is what we call the dynamic instability
axonemes  The use of the dynamic instability of the
Key Maintain cell’s shape and polarity;
functions microtubules allows the cell to rapidly
provide track for organelle and organize the cytoskeleton when necessary
chromosome movement; move cilia
 Dynamic microtubules are individually short
and flagella
lived, so arrays of microtubules are continually
in the process of recreation
The Dynamic Instability of Microtubules
 Because the microtubule shrinkage and growth
are active processes, it consumes energy and
this turn over can be fast, on the order of
minutes. This means that an array of
microtubules can adapt quickly to changes in
the environment, adopting new spatial
arrangements in response to cellular needs.
Examples include the reorganization of the
cytoskeleton in the transition to mitosis and
the extension of growth cones from neurons.

NUCLEUS  Take care of the DNA, DNA can be found in

the chromatin, chromatin is a strand of DNA
wrapped around histones
 Nuclear pores – where mRNA can transfer to
hormones/steroid inside and attach to the
nucleus
 Central Dogma (The 'Central Dogma' is the
process by which the instructions in DNA are
converted into a functional product)
o Transcription
 Center or brains of your cell o Translation
 Control center o Protein synthesis
 Dictates what proteins are to be done, proteins  Function of nucleus
are biomolecules that make up for the structure o Cellular regulation
of the cell secretions, the products, and their o Houses genetic material
overall structure and physiology o Directs cellular activities and
production of ribosomal units in the
nucleolus
Histology lecture: Cytoplasm 4
Nuclear Envelope
 Protects the DNA form the rest of the
cytoplasm
 Selective
 Cytoplasm is separated from nucleoplasm by
the nuclear envelope, a double set of
membranes with a narrow perinuclear space;
the outer membrane binds ribosomes and is
continuous with the RER.
 Central Dogma
1. Transcription – open the double helix
of the DNA, transcribe a part and
convert I to an RNA. The RNA from
the nucleus goes outside to the
cytoplasm and meets with your
ribosomes to create your protein
2. Translation – translate your amino
acids forming a peptide.
Peptide – combination of your amino
acids
Polypeptide – other name for protein
 The nuclear envelope is penetrated by nuclear
pore complexes, large assemblies of
nucleoporins with eightfold symmetry through
which proteins and protein–RNA complexes
move in both directions.
 The nuclear envelope is supported internally
by a meshwork, the nuclear lamina, composed
of intermediate filament subunits called
lamins.
*Green = DNA
*Red balls = histones
DNA wraps around histones creating a chromatin
Chromatin – colors the nucleus
Common staining in laboratory are hematoxylin
and eosin. Hematoxylin is an alkaline stain, binds
with acidic compound, a making the nucleus color
blue. Eosin, an acidic stain which binds to alkaline
compounds, usually proteins are alkaline. Proteins
are commonly seen in your cytoplasm. This
explains why under the microscope, nucleus is
blue and the cytoplasm is red. The DNA makes
your nucleus blue.
 Several orders of DNA packing occur in
chromatin and during chromatin condensation
of mitotic prophase. The top drawing shows
the 2-nm double helix, followed by the
association of DNA with histones to form 11-
nm filaments of nucleosomes connected by the
DNA (“beads in a string”). Nucleolus on the
DNA then interact in a manner not well
understood to form a more compact 30-nm
fiber. DNA in such fibers form various loops,
some of which in euchromatin involve gene
transcription. Loop remains tethered to and
stabilized by interactions with protein
scaffolds that eventually make up a central
framework at the long axis of each
chromosome. Heterochromatin is not
transcribed and remains more highly
condensed. The bottom drawing shows a
metaphase chromosome, with the maximum
packing of DNA. The chromosome consists of
two chromatids held together at a constriction
called the centromere.
Histology lecture: Cytoplasm 4

form nucleosomes, producing a structure

resembling beads on a string.
 Additional levels of chromatin fiber
condensation are less well understood and
involve nonhistone proteins, including
complexes of condensins.
 The extra X chromosome in cells of female
mammals forms facultative heterochromatin
and can be seen as the Barr body. Barr bodies
are can be seen in the neutrophils and oral
mucosa of females.
Types of heterochromatin
1. Constitutive heterochromatin
is generally similar in all cell types and
contains mainly repetitive, gene-poor DNA
sequences, including the large chromosomal
regions called centromeres and telomeres,
which are located near the middle (most often)
and at the ends of chromosomes, respectively.
Constitute to the structure of th chromatin
Poor in transcription
2. Facultative heterochromatin
contains other regions of DNA with genes
where transcription is variably inactivated in
Types of chromatin:
different cells by epigenetic mechanisms and
1. Heterochromatin – condensed can undergo reversible transitions from
- Inactive chromatin compact, transcriptionally silent states to more
- Needs to unravel through open, transcriptionally active conformations.
transcription to be There a chance of transcription
euchromatin
2. Euchromatin – active chromatin
Nucleoli
Chromatin will be condensed and become
heterochromatin, this will eventually become
chromosome. This is because mitosis is compact
Chromatin

 Chromatin is the combination of DNA and its

associated proteins.  Always there
 Chromatin with DNA that is active in  Site for your RNA production
transcription stains lightly and is called
 In a normal staining you do not see the
euchromatin; inactive chromatin stains more
nucleoli but if the cell is very active
darkly and is called heterochromatin. The
physiologically, increase in cellular, and
DNA molecule initially wraps around
protein synthesis activity, them most of the
complexes of basic proteins called histones to
chromatin will be converted into euchromatin.
Histology lecture: Cytoplasm 4

If it is euchromatin is present you can see the phosphorylate and activate the enzymes and
nucleoli. But usually can be seen because it is transcription factors whose functions
mixed together with heterochromatin characterize each phase of the cell cycle
o S phase – is the DNA synthesis
TEM reveals morphologically distinct region
o DNA now is trying to duplicate itself
within a nucleolus, Small, light-staining area are
in preparation for the next mitosis.
fibrillar centers (FV), containing DNA sequences
o G2 phase is cytoplasmic preparation
for the rRNA (the nucleolar organizers). The
darker the fibrillar material (F) surrounding the for mitosis
fibrillar centers consists of accumulating rRNA  Progress through the cell cycle stages is
transcripts. More granular material (G) if the monitored at checkpoints, including the G1
nucleolus contains mainly large and small restriction point; only when each phase’s
ribosomal subunits being assembled from rRNA activities are completed are the cyclins
and ribosomal proteins synthesized in the changed to trigger those of the next phase.
cytoplasm. Various amounts of the
heterochromatin (H) are also typically found near
the nucleolus, scatters in the euchromatic €, and
adjacent to the nuclear envelope (NE) that
separates chromatin from cytoplasm (C). (x35,000)
Nucleolus

 The nucleolus is a very basophilic or electron-

dense area of chromatin localized where rRNA
transcription and ribosomal subunits assembly
Quiescence - rest
occur.
Differentiation – mature
 By TEM, an active nucleolus is seen to have
Telophase – cleavage formation
fibrous and granular parts where rRNA forms
and ribosomal subunits are assembled, Mitosis
respectively.
The Cell Cycle

 The cell cycle is the sequence of events that

controls cell growth and division.
 The G1 phase, the longest part of the cycle,
begins immediately after mitosis and includes  Stages of mitotic cell divisions include
all preparations for DNA replication. o prophase, when chromosomes
o G1- Trying to recuperate after condense, the nuclear envelope
division, fill up those cells disassembles, and the microtubular
cytoplasmic structures and organelles spindle forms;
– nutritionally dependent o metaphase, when chromosomes are
 The period of DNA (and histone) synthesis is aligned;
the S phase. In a short G2 phase the cell o anaphase, when they begin to separate
prepares for division during mitosis (M). Cell toward the two centrosomes; and
cycling is controlled by the sequential o telophase, when nuclear envelope re-
appearance of key cytoplasmic proteins, the forms around the separated
cyclins, which bind CDKs. CDKs chromosomes.
Histology lecture: Cytoplasm 4

 Telophase ends with cytokinesis or cell more rapidly than stem cells before slowing or
cleavage into two daughter cells by a stopping division to differentiate.
contractile ring of actin filaments and myosin.

 In tissues with stable cell populations (mostly

 From stem cells, it will undergo mitosis, one
of the 2 daughter cell will mature and one will
differentiates and become columnar cell lining
in the intestine.
 Stem cells purpose is to replenish the
population
 In rapidly growing adult tissues and perhaps in
other tissues, there are slowly dividing
populations of stem cells. Stem cells divide
asymmetrically, producing one cell that
remains as a stem cell and another that
becomes committed to a differentiative
pathway but divides a few more times at a
more rapid rate. Such cells have been termed
progenitor cells, or “transit amplifying cells”,
each of which eventually stops dividing and
becomes fully differentiated.
Stem Cells & Tissue Renewal
connective tissue), such as most connective
tissues, smooth muscle, and the cells lining
blood vessels, stem cells are not readily
apparent and differentiated cells appear to
undergo slow and episodic division to
maintain tissue integrity.
 Labile cell – cell that keeps on dividing
 Permanent cell – stem cell that will
differentiate immediately. Examples of this are
neurons and muscle, meaning they don’t
undergo mitosis anymore
 Differentiate cells that have become stable
cells can remain stable or undergo meiosis.
 Stable cells - hepatocytes
Meiosis

 Stem cells occur in all tissues with rapid cell

turnover; they divide slowly in an asymmetric
manner, with one daughter cell remaining a
stem cell and one becoming committed toward
differentiation.
 Meiosis is the process by which two
 Cells committed to differentiate (transit successive cell divisions produce cells called
amplifying or progenitor cells) typically divide
Histology lecture: Cytoplasm 4

gametes containing half the number of

chromosomes found in somatic cells.

 Prophase of the first meiotic division is a

unique, extended period in which homologous
chromosomes pair and undergo genetic
recombination during the process called
synapsis. Synaptic pairs separate toward two
daughter cells at the first meiotic division.

 The second meiotic division occurs with no

intervening S phase and separates the sister
chromatids into two final cells that are haploid.
Mitosis -46
Meiosis - 23
Apoptosis

 Apoptosis is the process by which redundant

or defective cells are rapidly eliminated in a
manner that does not provoke a local
inflammatory reaction in the tissue.

 Apoptosis involves a cascade of events

controlled by the Bcl-2 family of proteins
regulating the release of death-promoting
factors from mitochondria.

 Cytochrome c from mitochondria activates

cytoplasmic proteases called caspases, which
degrade proteins of the cytosol, cytoskeleton,
and cell membrane.

 Endonucleases are activated, which degrade

all nuclear DNA. Cell and nuclear volumes
shrink rapidly, and the cell membrane changes
produce extensive blebbing of the cell surface.

 Late in apoptosis, the cell breaks into many

small apoptotic bodies that undergo
phagocytosis by neighboring cells.

 Apoptosis occurs rapidly, with little or no

release of proteins that would trigger
inflammation, unlike the death of injured cells
by necrosis that typically induces local
inflammation