Micro Lecture Review I

Rachel Boggus
boggusrl@email.uc.edu
419.376.9697
 Lecture Review

• H&E
– Hematoxylin
basophilic thingsblue
– Eosin acidophilic
thingsred/pink
• When would you get
cytoplasmic
basophilia?
• PAS (periodic Acid Schiff)
– Stains things bright
magenta
– What does it stain?
Answers
• H&E
– Hemotoxylin basophilic thingsblue
• When would you get cytoplasmic basophilia?
– Protein synthesizing cells, lots of ribosomes
basophilia
– Eosin acidophilic thingsred/pink
• PAS (periodic Acid Schiff)
– Stains things bright magenta
– What does it stain?
• Think of the G’s: Glycogen, goblet cells,
glycocalyx
Lecture review (cont)
• Transmission EM
– Normal EM
• Scanning EM
– Coat surface with metal, lets you see
surface anatomy really well
• Freeze Fracture
•You should
definitely be
able to tell
what type of
EM is in use in
each slide
•Remember
that Freeze
Fracture uses
TEM to view.
–This will be
on your test
fo sho
Membranes
• Unit Membrane
– Dark-light-dark
• Gives you an idea of the magnification
of an EM
– How is plasma membrane diff. from
others?
– Which 2 organelles are surrounded by 2
membranes?
– Know the difference btwn integral memb
proteins and peripheral memb proteins
– What 4 lipids make up the plasma
membrane?
• Unit Membrane
– Dark-light-dark
• Gives you an idea of the magnification of an EM
– How is plasma membrane diff. from others?
• Contains 5% carbohydrates (glycoproteins)
– Which 2 organelles are surrounded by membranes?
• Mitochondria and nucleus
– Know the difference btwn integral memb proteins and
peripheral memb proteins
– What 4 lipids make up the plasma membrane?
• Phosphotidylcholine, phosphatidylethanolamine,
phosphatidylserine, and sphingomyelin
– All of them are amphipathic
Membranes (cont)
• Lipid Asymmetry
– Which two phospholipids are on the outer
surface?
• What are the consequences of this
arrangement?
– What is the role of cholesterol?
– Lipid Rafts—just know them
• Rich in sphingolipids and cholesterol
• Assoc. with calveolin and GPI linked proteins
• Play a role in membrane sorting—how some
toxins get in cell
• Detergent insoluble
– Which two phospholipids are on the outer surface?
• Phospohtidylcholine and sphingomyelin
• What are the consequences of this arrangement?
– Charge difference between inner and outer leaflets
because phosphatidylserine has a negative charge.
• What is the role of cholesterol
– Restricts mobilityinhibit phase transition
RBC Memb proteins
• Name them
• What do they do
RBC Memb proteins
• Name them
– Spectrin, Glycophorin, Band III, Ankyrin
• What do they do
– Spectrin—RBC cytoskeleton (peripheral)
– Glycophorin—heavily glycosylated
(integral)
– Band III—bicarbonate/chloride exchanger
(integral)
– Ankyrin—links spectrin and band III
(cytosolic)
Here they are….

I know. Shit picture labels. Sorry.

Glycoproteins, glycolipids, and glycocalyx

• Terminate in sialic acid negative

charge
– Sugar arrangement is ordered.
– You may as well learn this now because
I’m pretty sure you learn how they are
made later
• Glycocalyx
– Carbs on outer leaflet of Plasma memb.
• What would you use to stain it? PAS.
– Participates in extracellular functions
More fabulous membranes
• Know the difference between active
and passive transport
– What can diffuse through plasma
membrane?
• Uniport, Symport, and antiport carrier
proteins
– Define, give example of antiport
 More fabulous membranes

• Know the difference between active and

passive transport
– What can diffuse through plasma membrane?
• Small hydrophobic molecules, H20
• Uniport, Symport, and antiport carrier
proteins
– Define each and give an example of antiport
• Uniport single ion in one direction
• Symport two ionic species the same direction
• Antiport two different ions in opposite directions
– Band III and Na/K ATPase
Endocytosis
• Phagocytosis
– Large stuff
• Pinocytosis
– Smaller stuff
• Receptor Mediated Endocytosis!
– Understand why it is saturable
– The role of clathrin
– Know the possible routes—transcytosis
etc.
Caveolae
• Dr. Morris loves stupid clathrin and caveolae
– Abundant on smooth muscle and endothelial cells
• Tuck that away in your long term memory because
that will help you differentiate Sm. Muscle and
endothelial cells in EM
– Definitely know that caveolae are associated with
lipid rafts and that it is involved in internalization
and cell signaling.
ER of the rough and smooth variety
• What are
the 3
classes of
proteins
made in the
RER?
• Where are
other
proteins
made?
• Name at
least 4 roles
of the SER?
• What are the 3 classes of proteins made
in the RER?
– Lysosomal enzymes, secretory
proteins, integral membrane proteins
• Where are other proteins made?
– Polysomes in cytoplasm
• Name at least 4 roles of the SER?
– Cholesterol metabolism, steroid biogenesis,
drug detox, glycogen metabolism,
phospholipid biosyn
• Note that A LOT of these take place in
hepatocytes
RER protein synthesis
• mRNA contain prepiece/signal
peptide
• Singal recognition particle (SRP)
– Binds signal peptide and transports complex to
the RERthreads sequence through forming a
loop sythesis resumes, SRP detaches
• In RER prepiece is cleaved and the protein is
folded and glycosylated
• N-linked glycosylation ASN-X-serine or
threonine
– Also dependant on dolichol phosphate
• Know there are chaperones
– Ubiquitinationdestroying misfolded ones
The Nucleus

• Nuclear envelopetwo unit membs.

– Underlying nuclear lamina
– Nuclear pores allow traffic btwn cyto and nuc.
• Nuclear localization sequence Lys-Lys-Lys-Arg-Lys
– This was on our test and I’m not even kidding
• importins bind NLS and import
• Export similar in theory (NES on proteins)
– Nucleolus– site of RNA synthesis
• Know that Chromatin is packaged DNA and
proteins (histones)
• Nucleosome (histone octamer and DNA)
basic unit of chromatin
•Heterochromatin is?

•Euchromatin is?

•A cell with a lot of

Euchromatin is…?
• Heterochromatin is?
– Stains darkly, transcriptionally inactive
– Divided into constituative and facultative
• Euchromatin is?
– Stains lightly, is transcriptionally active
• A cell with a lot of Euchromatin is…?
– Doing a lot of protein synthesis
THE CELL CYCLE/MITOSIS
• If you don’t
know it, learn
it.
• G1, S, G2, M
• Prophase
• Metaphase,
Anaphase,
Telophase,
Cytokinesis
• Differences
between
mitosis and
meiosis
Golgi Apparatus
• Modification and distribution center of
cell
– Carb modification
• What are the 5 regions of the Golgi,
what direction does crap travel?
• Know the diagram about the carb
modification
• Modification and distribution center of
cell
– Carb modification
• What are the 5 regions of the Golgi,
what direction does crap travel?
– Cis-golgi networkcis regionmedial
regiontrans region trans golgi network
• What identifies lysosomal enzymes?
• What are other functions of the golgi?
• What identifies lysosomal enzymes?
– Mannose-6-phosphate
• On our test. Know it.
• What are other functions of golgi (4)?
– Lipoprotein packaging, sulfation,
proteolytic processing of proproteins, and
assembly of lipid rafts (remember
these???)
Proproteins
• Have an extra piece compared with
actual protein
• Remember that originally it was a pre-
pro-protein, prepeice cleaved in RER,
transported to GA, propeice cleaved
during transport.
– Remember clathrin?
• It takes lysosomal enzymes to lysosomes
• REGULATED SECRETION
• What happens to proteins that don’t
have a specified delivery point?
• What happens to proteins that don’t
have a specified delivery point?
– They are routed to the cell surface and
released constitutive secretion (Igs)
Vesicular Transport
• Unidirectional
– Budding and fusion
– Understand the diagram in your notes
Realize that there are 2 models of
progression through golgi
– Don’t confuse COP I and COP II
• What are the 3 features shared by all
lysosomes

• Don’t forget about mannose-6-P

• What are the 3 features shared by all
lysosomes
– Limited by single unit membrane
– All are acidic
– All are acid phosphatase positive
• Don’t forget about mannose-6-P
Mitochondria
• How do you distinguish a
mitochondria?
•How do you
distinguish a
mitochondria?
–Surrounded by 2
membranes
–Contain cristae
arising from inner
membrane
Peroxisomes
• How can you
tell?
• Compare it
to the dark
lysosome at
the top, that
would be a
secondary
lysosome
 Image Review #1
Labs 1-3

GO BLUE
 LAB #1

“THE CELL”
 STAINS
• H and E
– Hematoxylin is blue/basophilic
– Eosin is pink/eosinophilic/acidophilic
 PAS
• Stain stuff that starts with G:
glycocalyx, goblet cells, glycogen,
glycoprotein
 PLASMA MEMBRANE
There are two of them here, each is trilaminar

Each is 9nm thick – can’t see well with LM

unless juxtaposed (needs to be at least 200
nm to see with LM, FYI)
 RIBOSOMES
• Stain BASOPHILIC (baso = blue)
• If you see bluish stuff in the cytoplasm that means
the ribosomes are busy and the cell is making
proteins

• Polysomes = ribosomes on an mRNA string

– Look like little swirls
 Golgi Apparatus

• Cis face and trans face – stuff buds off

of the trans face so trans face is more
DILATED
• Looks like a smiley face – trans face is
on the upper lip and cis face is on the
chin
• Cis face faces nucleus, trans face faces
the outside of cell – makes sense when
you think about how things are
transported
Meet the EM Golgi – look at the
budding vesicles!
Don’t forget about the golgi ghost
in LM slides!!

BOO!!
 Mitochondria
• Via oxidative phosphorylation, supply ATP for
the cell

• Cristae? Mg and Ca granules? What other

organelle do we see here?
 ER
• Made of cisternae – flattened tubes
• Two types: RER and SER
– Can they be connected? YES!
– The difference? RER has ribosomes, SER
does not

If you see cytoplasmic basophilia in a

LM slide, then there is a lot of RER in
the cytoplasm. The basophilia is due
to the rRNA of the ribsomes that are
attached to the RER.
Hello there basophilia
 ER cont’d
If you are talking about neurons,
cytoplasmic basophilia is called
“ergastoplasm” and the RER stains
with Nissl stain. Thus, the RER is
called Nissl substace. Makes sense.
Neuron RER = nissl
 SER
• Usually rounder than RER and looks
like it has more buds on it. Also, no
ribosomes. That always helps.
• SER SECRET: if you think it may be SER
or RER or Golgi or you have no clue,
and you can’t decide what to put
down, look for the mitochondria if you
can. If the mitochondria are really
round and circular, there is a good
chance that it is SER.
– this is because in cells that are involved in
detox functions (and thus have lots of
SER), the mito are rounder.
SER abundance
RER vs. SER
RER in plasma cell
• Also notice lots of euchromatin in the
nucleus – this cell is clearly active,
making Igs to be exact
SER
 Lysosomes
• Have a membrane, UNLIKE
peroxisomes, which do not
• Contain acid phosphatases and acid
hydrolases to break stuff down – can
stain for these enzymes to find the
lysosomes
Primary has no crap in it
Secondary has crap in it

CRAP
 Peroxisomes
• Very smooth looking except for a huge
granule (not in humans though, just
rats)
• Contain catalase and oxidase
peroxisomes
 Cytoskeleton
Microtubules
Microfilaments
Intermediate Filaments
Microtubules
(24 nm)
Don’t forget…. They are involved
in the mitotic spindle
 CENTRIOLES
• Involved in cell division
• Can you find it?
One more just for good measure
 Microfilaments
-- Made of actin
-- Contractile – look for them in muscle
-- 7nm – smallest of the cytoskeletal
components
 More microfilaments
• They also make up the core of
microvilli
 INTERMEDIATE FILAMENTS
• 10 nm – intermediate in size between
microfilaments and microtubules
• They are made of protein that are
tissue specific
• A main component of desmosomes……
CELLULAR INCLUSIONS
GLYCOGEN
PIGMENT/MELANIN
LIPID
 GLYCOGEN
• Solid and black in EM
• Reddish in LM, but you will usually get
EM if they ask you about glycogen
LM glycogen from lab
LIPID
 More lipid….

Also note the round mito….. What do you see with round
mito??? SER. One of SER’s functions is fat metabolism, so
makes sense that there is lipid there too.
 PIGMENT
• aka melanin, so if they are trying to
show you pigment it will usually be in
the skin, in melanocytes, which are
found under the keratinocytes.
Secretory/zymogen granules
• It is what it sounds like. Not brain
surgery here.
 The nucleus
In all its glory
 Interphase nucleus
• Has nuclear membrane – 2 membranes juxtaposed
with pores in them

• Note: nuclear membrane made of intermediate

filaments
Taking a peek into the nucleus
• Heterochromatin – dark, bundled up
chromatin, inactive
• Euchromatin – light, unwound, ready
to go, active
 Nucleolus
• 3 things they could label:
– Perinuclear chromatin – chromatin pressed
onto the outside of the nucleolus
– Fibrillar portion – the darker stuff
– Granular portion – the lighter stuff
 Prophase
• Nuclear membrane disrupted
• Chromatin looks fibrous, stringy
– It is condensing to become chromosomes
 Metaphase
• Chromosomes are condensed and lined
up in the middle, the mitotic spindle is
evident
• The mitotic spindle converges at
centrioles – called the “Microtubule
Organizing Center” or the centrosome
– 3 types of microtubules here
• 1. Astral MT that radiate out like a star – help
with orientation
• 2. Mitotic spindle MT that go across the spindle;
can see these ones the best
• 3. Microtubules that are attached to the
chromosomes
Metaphase
Anaphase
• Chromosomes are separating
• Can clearly see the spindle and astral
microtubules
 Telophase
• Can start to see new cell membrane
between the 2 new daughter cells
• Chromosomes look more separate
again – not as distinct as in anaphase
 Karyotype
• This was on our exam and a lot of
people forgot what it was called. Do
not be that guy.
 Quick Quiz
What is the main structure, and also the
cytoskeletal element that you see here?
Cilia, with a core of microtubules!
Whats the black stuff??
Glycogen!
What are the arrows pointing to?
Microtubules in the core of a
dendrite!
What is the dark organelle shown
here?
Lysosomes! Look at all the stuff in
it! That would make it secondary,
yes?
Whats this??
 On the left is the Golgi
On the right is a centriole
What is found in these cells?
Melanin! The arrow is pointing at
the row of melanocytes under all
the keratinocytes in the skin
Whats this??
NUCLEAR PORE!
 The End
Questions???
Boggusrl@email.uc.edu