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Nucleus

● largest organelle of the cell


● contains nearly all DNA; minute in cytosol
● RNA synthesis: nucleolus
● Varies in shape
○ Spherical- squamous cells
○ Elongated (oval)- columnar cells
○ Globulated/Segmented- neutrophils
● Varies in number within the cell
○ No nucleus- mature RBC
○ Uninucleated/ Mononucleated
– cuboidal cells of kidney
– columnar cells of small intestine
– squamous cells
○ Binucleated- cuboidal cells of liver (hepatocytes)
○ Multinucleated- skeletal muscle cell

Nuclear Envelope/Membrane
– encloses the nucleus
● barrier to ions, solute, macromolecules
● two parallel cellular membrane
● Perinuclear cisternae- space between the two membranes
● Outer membrane
– attached to RER
– ribosomes
● Inner membrane
– meshwork of fibrous proteins
– supported with lamins/lamina
– type of intermediate filament
● Membranes fused to form pores
○ complex assemblies of proteins
Within
● Chromosomes- condensed
● Chromatin- uncondensed; coiling/ condensation
● Nuclear Matrix- fibrillar network
● Nucleoli- rRNA and ribosomes
● Nucleoplasm- fluid of nucleus

Nuclear Pore Complex


– fusion point of the two membranes
● Huge macromolecular complex
● Nuclear pore + glycoproteins
● 80-100 nm in diameter
● Composed of 3 ring-like arrays of protein
○ Rings are on top of one anoter
● Octagonal symmetry
○ Each ring (cytoplasmic and outer): 8-fold repetition of subunits
○ 30-50 nucleoporins (protein)
● Nucleoporins
○ Symmetrical on both cytoplasmic and nuclear sides
● Low molecular with solutes- diffuse freely
● Macromolecules- regulated, mediated by receptors, energy-requiring
● Bidirectional transport mechanism (entry and exit)
Rings
● Cytoplasmic Ring
○ filamentous protein (cytoplasmic filament/ran-binding filament)
– attract molecules: move along the primitive before crossing NPC
● Middle Ring
○ anchors NPC to perinuclear ciserna
○ stabilized NPC
○ hour glass structure: central transporter
– gated channel; allow small molecules to cross NPC
● Nucleoplasmic Ring
○ Nuclear basket
– flexible part of NPC; depends on the size of the molecule entering
◆ low molecular substances; less than 80 nm
◆ <100- macromolecules; entry is regulated; energy requiring; aided by
receptors

● Transport Receptors- regulate the entry of big molecules; soluble


● Exportins
○ nucleus to cytoplasm
○ transports RNA (RNA is inside the cell)
● Importins
○ cytoplasm to nucleus
○ proteins from ribosomes
○ more substances
○ has alpha and beta subunits
– bind to specific protein; amino acid sequencing
– run along cytoplasmic filament and enter gated channel
– energy requiring: GTP
– beta will be left outside the nucleus; alpha will enter with protein
– Exportins will take the protein, GTP, and load outside
– protein is outside: GTP and exportins can re-enter nucleoplasm

Chromatin
● seen when cell is in interphase
○ not resting; preparing for cell division
● guide to the cellʼs activity
● Euchromatin
○ more abundant in the center; interior
○ regions of DNA containing active genes (dispersed, active DNA)
○ lighter appearance, paler appearance
○ L.M. : lightly stained basophilia, lightly bluish
○ E.M. : beads on string
◆ nucleosomes- beads
◆ naked DNA- string; coiled around to conserve DNA
◆ stabilized by duplicates of 4 types of Histones
◆ octomeric
◆ bonds on string appearance: H1
◆ nucleosome core particles:
◆ H2A
◆ H2B
◆ H3
◆ H4
◆ 164 bp in loop
◆ 6 to 7 fold shortening
◆ 10 nm fiber
● Heterochromatin
○ Inactive, condensed chromatin
○ located at the edges of the nucleus
○ visible by E.M. as a darker colour
○ L.M. : Basophilic clumps
○ Stains deeply with Feulgenʼs stain
○ Two types:
◆ Constitutive heterochromatin- non-coding; can never become active
◆ Constitutive (structural parts):
◆ centromere- center
◆ telomere- end
◆ Facultative heterochromatin
– contains genes that are shut off and temporarily inactive
◆ In any cell only a portion of all the genes are active
– Example: the Barr Body. Females have two X chromosomes, one
of which is inactive

Nucleoplasm
● Consists of:
○ interchromatin granules (IGs)
○ perichromatin granules (PCGs)
○ ribo-nucleus protein
◆ plus the nuclear matrix
● Nuclear Matrix
○ 10% CHONs
○ 30% RNA
○ 1-3% DNA
○ 2-5% PO4-3
● Nucleolus
○ Spherical
○ Highly basophilic
○ Active in protein synthesis
○ Non-membrane bound structure
○ Assembles 40s and 60s ribosomal subunits

Cytoskeleton
Functions:
● Structural support
● Internal framework maintaining position of the organelles; 3D shape
● Cell motility; machinery required for movement of materials and organelles
within cells
● Force generating elements responsible for movement of cells from one place
to another

Three types:
Microtubules
● Ultrastructure; thickest
● Hollow
○ cylindrical with lumen
◆ free space in the interior aspect of the structure
● Tubular structures 25nm in diameter
● Forms the mitotic spindle, centrioles, core of cilia and flagella
○ mitotic spindle
– during cell division, centrioles will move to the opposite pole of the cell;
from centrioles mitotic spindle will arise
– guide the chromosomes in the alignment process (equatorial plane;
metaphase)
– made up of microtubules
● made up of 13 rows of protofilaments arranged in a circle
○ 1 strand: alternating alpha-beta tubulin heterodimers
● have a plus and minus end
○ plus end- addition of tubulins occur; lower microtubules: alternating
alpha-beta
○ minus end- stabilized, no addition of proteins
● Microtubule Associated Proteins (MAPs)
○ increase the stability and promote microtubule assembly
○ avoid displacement of alpha and beta tubulins in the protofilaments
● Motor proteins: kinesins, dyneins move along microtubules
○ All require ATP for movement; active transport mechanism
○ Move along microtubules in opposite directions
◆ Kinesins- transport organelles toward the plus end
◆ Dyneins- tranport proteins toward the minus end
● Provide the basis for several complex cytoplasmic components:
○ Centrioles
◆ 9 microtubules arranged in triplets
◆ Cell division: centrioles replicate
◆ Paired: centrosome (where mitotic spindle arise)
– no direct connection; right angles with each other;
– pairing is stablized by a sticky material: pericentriolar material
◆ MTOC (Mictrotubule Organzing Center)
◆ ABC microtubule
◆ A- nearest to the center
◆ C- farthest
◆ All As are connected to the C tubule of the adjacent triplet
◆ All the As are connected to the center: radial spoke
◆ Centrioles are in pairs and at right angles to each other.
● Central structural supports in cilia and flagella.
– Both can move unicellular and small multicellular organisms by propelling
water past the organism.
– If these structures are anchored in a large structure, they move fluid over a
surface.
○ Cilia
◆ Uncheated structure
◆ Sweep away water and dirt on the surface
◆ Cilia usually occur in large numbers on the cell surface.
◆ 0.25 microns in diameter and 2-20 microns long.
◆ Move with alternating power and recovery strokes.
◆ Movement is perpendicular to the axis
○ Flagella
◆ Provide motility; locomotory structure
◆ One or few per cell
◆ Same width as cilia
◆ Longer than cilia
◆ 10-200 microns long.
◆ Undulatory- force is parallel to the axis of the flagellum
○ Both have a core of microtubules sheathed by the plasma membrane.
○ Nine doublets of microtubules arranged around a pair at the center, the “9
+ 2” pattern.
◆ Made up of 9 doblex of micrtubules plus a pair at the center
◆ Doblex connected to each other via dynein arms.
◆ Doblex are connected to the pair at the center: radial spoke
○ Flexible “wheels” of proteins connect outer doublets to each other and to
the core.
○ The bending of cilia and flagella is driven by the arms of dynein.
○ Have a covering/plasma membrane

Microfilaments/ Actin Filaments


● Thinnest
● Designed to resist tension
● Solid rods of the globular protein actin.
● Made up of twisted double chain of actin subunits
○ No lumen
○ 2 strands/filaments
◆ each strand: F actin (filamentous)
– consists of globular structures: G actin (globular)
◆ F actin- composed of connected G actins
◆ Microfilaments- 2 F actins twisted together
● Arranged parallel to one another (muscle cells)
● Thicker filaments, composed of a motor protein, myosin, interdigitate with the
thinner actin fibers.
● Myosin molecules walk along the actin filament, pulling stacks of actin fibers
together and shortening the cell.
Function
● Anchor the microvilli to the underlying intermediate filaments
○ microvilli- foldings intestine, increase absorptive capacity of small
intestine, increase surface area: foldings
– supported by microfilaments
– underneath the actin filaments: intermediate filaments
● Muscular contraction
● Actin-myosin interactions and sol-gel transformations: cytoplasmic streaming.
– creates a circular flow of cytoplasm in the cell.
– speeds the distribution of materials within the cell.; even distribution of
materials
● Purse-ring (indentation) to accommodate the engulfing of substances
● Stress fibers in crawling cells
○ crawling cells: neutrophils (can escape blood vessels)
◆ actin filaments allow neutrophils to form pseudopods
● Separate the parent cells into two daughter cells during cytokinesis (cell
division process)
○ Cleavage furrow (telophase stage)
– Actin filaments will concentrate here and contract until complete
separation of cytoplasm occurs

Intermediate Filaments
● Thickness is between microtubules and microfilaments
● Designed to bear tension
● Reinforce the 3D shape of the cell
● Fix organelle location
○ Organelles are not displaced in the cytoplasm because they are anchored/
stabilized because of the intermediate filaments
● Are not scattered

4 major groups
● Keratin
– found in the skin
– prevent dehydration
● Vimentin
– found in the mesenchymal cells (undifferentiated cells)
● Neurofilaments
– supports axon and dendrites (nerve cell)
● Lamin
– Outer and and inner membrane of the nucleus is supported by crisscross
proteins: nuclear lamina
– made up of intermediate filaments

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