Cell Biology 1:

Membranes, Cytoplasm and Nucleus

Medical Microanatomy 602
Edie C. Goldsmith, Ph.D.

M=mitochondria ER=rER N=nucleus NE=nuclear membrane
G=golgi apparatus V=vesicles PM=plasma membrane
C=neighboring cell IS=intercellular space L=lysosome
See Table 2.1 pg 24-25 in your textbook
Plasma Membrane
8-10 nm thick
Function
Selective barrier
Cell adhesion
Communication
Three main components
Phospholipids
Proteins (integral or
peripheral)
Cholesterol


Modified Fluid Mosaic Model
Lipids
Membranes usually have
40-60% lipid
Type varies
Phosphatidylcholine,
phosphatidylserine,
phosphatidylethanolamine
glycolipids
Polar and non-polar
groups
Saturated vs unsaturated
Not uniform distribution
rafts
Lipid Rafts
Enriched in Cholesterol and
Sphingolipids

Less fluid than typical membrane
Due to the
amphipathic nature
of lipids - self
associate
Micelles
Bilayers
Lipids can move
within a bilayer

Cholesterol
Integrates into bilayer
Amount in PM can vary
by cell type


Modulates membrane
fluidity

Membrane Proteins
~50% of membrane
(w/w)
Two general types
Integral
Peripheral
Proteins are also
asymmetrically
distributed with respect
to the lipid bilayer
Multiple categories of
integral membrane
proteins
Pumps and enzymes (ion
pumps)
Channels (gap junctions)
Receptors
Linker (integrins)
Carrier proteins
Structural Proteins
Glycoproteins and
glycolipids
glycocalyx
Freeze-fracture
The interior of the lipid
bilayer is weak and
readily split by simple
cleavage.
Membrane proteins will
stay with one or the
other bilayer forming
bumps and leaving pits.
Freeze Fracture
The two faces of the
cleaved lipid bilayer
may be examined using
freeze fracture to help
determine membrane
protein distribution.
Imaged by TEM
TEM vs. Freeze Fracture
Top: Gap junction
between plasma
membranes of two cells

Middle/bottom: same
structure viewed en face
using freeze fracture
Transport Mechanisms
Diffusion - Passive and Facilitated
Passive concentration gradient
Gases, lipids, lipid soluble molecules (i.e. EtOH)
Facilitated concentration dependent
Require protein carrier molecule; does not require energy
Glucose and amino acids (hydrophilic)
Active transport
Requires energy (ATP)
Movement against gradient
Na
+
-K
+
ATPase
Channel Proteins
Small, water soluble molecules
Proteins create pore in membrane; ion selective
Regulated by membrane potential (voltage-gated), neurotransmitters
(ligand-gated), mechanical stress
Transport Mechanisms
Vesicular transport
Endocytosis
Pinocytosis
Receptor mediated endocytosis
Phagocytosis
Exocytosis

Receptor-mediated Endocytosis
Receptor mediated
LDL, transferrin/Fe, EGF
Clathrin dependent
Assemble on cytoplasmic side
of membrane
Weave-like network
Coated pits
Fuse with endosomes
dynamin
Phagocytosis
Macrophages and
polymorphonuclear
leukocytes
Engulfment of large
particles
~ 250 nm
Bacteria, protozoa,
fungi, damaged cells
Non-biological debris
Clathrin independent
actin dependent
Receptor mediated
Fc receptors
Pinocytosis
cell drinking
Clathrin independent
Smooth vesicles
80-150 nm
Intake of fluid and
small molecules
Usually fuse with
lysosomes
Bulk transfer
(endothelial cells)
Exocytosis
Mechanism for
releasing molecules into
extracellular space
Two pathways
Constitutive
All cells
Secreted immediately; No
granules
Secretory
Specialized cells
Proteins to be secreted
stored in vesicles
Cytoplasm
Aqueous matrix containing organelles
About total volume of cell

Cytoskeleton

Site of many metabolic pathways
Glycolysis, protein synthesis
Nucleus
3-10 mm
Usually largest organelle
Function
Genome, production of
ribosomes and mRNA
Multiple nuclei
Not always in center of cell
Not always round
Multiple components
Chromatin
Nucleolus
Nuclear envelop
Nucleoplasm
Reticulocytes lose the nucleus entirely
to become mature RBCs
Muscle cells have multiple nuclei
Chromatin
DNA-protein complex
histones
Basophilic staining of
nuclei
Heterogenous
Heterochromatin - highly
condensed (dense
appearance)
Cell inactive
Euchromatin - dispersed
(light appearance)
Active cell
Some cells are very
inactive, and their
nuclei contain mostly
heterochromatin.
This is a late red blood cell precursor
Nucleus
The nucleus shown in
this image appears in
four parts.

Can cells have multiple
nuclei? Does this cell
have multiple nuclei?
Chromosomes
See Folder 3.1 pg 80-81
Nucleolus
No membrane
Basophilic
Three components
Fibrillar material/region (Pars
fibrosa) active transcription
of rRNA genes; lots of rRNA
Granular material/region (Pars
granulosa) ribosomal
assembly
Fibrillar centers regions of
DNA that code for rRNA,
RNA Pol I, transcription
factors
Nucleolar organizer region
NOR
Nucleoli are the site of
rRNA synthesis
May vary greatly in size
and appearance
Active cells may
contain multiple
nucleoli
Light microscopy of Primary Oocytes
Nuclear Envelope
Two membranes separated
by narrow space
(perinuclear cisterna)
Outer membrane
contiguous with rER
Inner membrane associated
with nuclear lamina
Lamins (A and C) provide
stability and association
points for chromosomes
Lamin mutations can affect
striated muscle, adipose
tissue, nerve/skeletal
development
Nuclear Pores
70 - 80 nm openings
Number and distribution
vary
Only site of direct
molecular exchange
with cytoplasm

Freeze fracture demonstrating distribution of nuclear pores
> than 50 subunits
Regulates bidirectional protein translocation
Ions & < 9 daltons freely
Large proteins/complexes, RNA, ribosomal subunits
require GTP
- Require nuclear localization signal (NLS) to get in
- Nuclear export sequence (NES) to get out

Cell Cycle
Cell population
Static - no division
Stable - sporadic
Renewing - regular
division
Slow (smooth muscle) or
rapid (dermal fibroblasts)
Cell Cycle
Mitosis
Interphase

Interphase
Three phases
G
1
Longest phase (9-12 hours)
Two checkpoints restriction point and DNA damage checkpoint
S
7.5-10 hours
DNA damage checkpoint fidelity of replication
G
2
3.5-4.5 hours
Cell growth and organelle organization
Two checkpoints Unreplicated DNA and DNA damage
G
0
/Terminal Differentiation
Mitosis
Four stages (~ 1hr)
Two checkpoints spindle
assembly (anaphase) &
chromosome segregation (prior to
cytokinesis)
Prophase
Chromosomes condense
Loss of nucleolus and nuclear
membrane
Metaphase
Assembly of mitotic spindle
Chromosomes migrate to
equatorial plane
Anaphase
Chromatids separate and migrate
toward poles
Telophase
Nuclear membrane and nucleoli
reappear; cytokinesis
I
P
M
A
Cell Cycle Regulation
Entry into different
phases is controlled
by cyclins and cyclin-
dependent kinases
(Cdks)
2-protein complex
Synthesis oscillates
up and down during
cell cycle
Apoptosis
Normal part of
development
Characteristics
DNA fragmentation
Decrease in cell &
nucleus size
Loss of mitochondrial
function
Membrane blebbing
Apoptotic bodies
Necrosis
Pathologic process
Characteristics
Damage to plasma
membrane
Swelling
Cell lysis
Inflammatory response
Apoptosis
vs Necorsis