TALBOT Q's [Membrane Biophysics & Resting Membrane Potentials, AP, Synaptic Transmissions, Actin Filaments, Muscle Contraction, The Neuromuscular Junction, Excitable Cells, Channelopathies] Flashcards
what stains actin?
rhodamine phalloidin
what stains tubulin?
anti-tubulin & fluorescein secondary antibody
what stains DNA/nucleus?
dapi
what’s the protein of actin filaments?
actin
what’s the protein of microtubules?
tubulins
what’s unpolymerized form of actin filament?
globular, monomer (G actin)
what’s unpolymerized form of MT?
heterodimer (1alpha +1beta)
what’s the bound nucleotide in actin filaments?
ATP
what’s the bound nucleotide in MT?
GTP
what’s the form of polymer in actin filaments?
2-stranded helix
what’s the form of polymer in MT?
hollow tube, 13 protofilaments
do actin filaments have dynamic instability?
yes
do MTs have dynamic instability?
yes
are actin filaments structurally polar?
yes
are Mts structurally polar?
yes
what’s the growing end of actin filament?
+ end
what’s the growing end of MT?
+ end
what’s the shrinking end of actin filament?
+ end
what’s the shrinking end of MT?
+ end
what are the roles of actin filaments?
- mircrovilli
- contractile bundles in cytoplasm
- form leading edge projections in crawling cells
- form the contractile ring in cell division
which end of actin filament do G-actin monomers bound to ATP add to?
+ end
which end of actin filament do G-acin monomers bound to ADP come off of?
- end
profilin
actin binidn protein that binds to actin monomers and facilitates exchange for ADP for ATP
treadmilling
refers to the fact that any given G-actin appears to “move along a treadmill” in that it adds on to the + end and falls off from the - end
TRUE or FALSE. “actin filament can grow or shrink at either end in vivo. however, they preferentially grow at their + end and shrink at their - end
True
ATP cap
as long as it is on the actin filament, the end in which it is located is where the actin filament grows
NOTE: if it comes off, ADP may be exposed, allowing possible shrinking at that end
severing proteins (aka. gelsolins)
they cut an actin in the middle, exposing ADP-actin on the plus end of the new fragment as well as on the new minus end; thus they promote rapid depolymerization and convert the gel-like structures into a more liquid solution
what is an example of cross-linking protein?
filamin
what are some examples of bundling protein?
fimbrin, villain, alpha-actin
what is an example of motor protein?
mysosin
what are some examples of capping (end-blocking) proteins?
capZ, tropomodulin
what is an example of side-binding/stabilizing proteins?
tropomyosin
what are some examples of nucleating proteins?
ARP 2/3, profilin
what do you call the filament growing out of ARP 2/3?
daughter filament
[difficult-long answer] what are the steps of cell crawling?
1) nucleating proteins promote pushing out actin filaments towards leading edge structures
2) bundling proteins in filopodia help form exploratory fibers
3) integrins form new adhesion point; severing proteins promote depolymerization of actin filaments
4) myosin’s walk along actin filaments, pulling rear of cell towards new adhesion point
motor domain of myosin
acts with actin and hydrolyzes ATP
what are the 3 distinct, fibrous components of the cytoskeleton?
intermediate filaments, MT, and actin
what is the similarity among the 3 distinct, fibrous components of the cytoskeleton?
they each is made of 2-13 protofilaments arranged in parallel to each other and held together by non-covalent bonds
what is the difference among the intermediate filament, MT and actin?
they each is unique in the type of protein subunits used to form each structure and in their individual roles in the cell
what are the similarities between actin and tubulin?
- bind and hydrolyze a nucleotide triphosphate and its relative form influences the affinity of the protein subunit for other subunits
- exhibit dynamic instability
- structurally polar, with functionally distinct ends
- the plus end preferentially grows whereas the mini send preferentially shrinks
what is the intracellular Na+ concentration?
15 mM
what is the interstitial Na+ concentration?
145 mM
what is the intracellular K+ concentration?
120 mM
what is the interstitial K+ concentration?
4.5 mM
what its the interstitial Ca2+ concentration?
1.2 mM
what is the intracellular Cl- concentration?
20 mM
what its the interstitial Cl- concentration?
116 mM
synaptic transmission is predominantly _____ (via NTs) but also can be _____ (via gap junctions).
chemical; electrical
in neurons that interact via _____ synapses, the pre-synaptic AP induces the secretion of NT, leading to a change in _____ in the postsynaptic cell.
chemical; Vm
in neurons that are connected by _____ synapses, the signal moves through gap junctions directly from the pre-synaptic cell to the post synaptic cell.
electrical
electrically coupled cells
cells that are connected by gap junctions
where can you find electrically coupled cells?
brain, cardiac muscle, and some smooth muscle
T/F in electrical transmission, there is some loss of voltage between the pre- and post-synaptic cell.
TRUE
T/F electrical synapses are almost always excitatory.
TRUE
what is responsible for the loss of voltage between the pre- and post-synaptic cell?
passive spread of a change in membrane potential
between electrical and chemical synapses, which one is only unidirectional?
chemical synapse
there are 9 steps in a chemical synapse. name all the 9 steps
1) NT synthesized and stored in vesicles in presynaptic cell
2) AP depolarizes nerve terminus
3) depolarization opens voltage-gated Ca2+ channels
4) Ca flows from ECF into cell
5) Ca acts as 2nd messenger, inducing vesicles to fuse with plasma membrane
6) NT released into synaptic cleft
7) NT binds to receptors on post-synaptic cell
8) this alters gating of ion channels in the post-synaptic cell
9) PSP (post synaptic potential )develops across post-synaptic cell dendritic membrane
SNARE proteins
proteins that allow the vesicle to recognize, dock and fuse to the plasma membrane; found in both vesicular membrane and palsmamembrane
which two synaptic toxins inhibit NT secretion?
botulism toxin
&
tetanus toxin
which synaptic toxin induces a massive non-specific release of NT?
alpha-latrotoxin
the vesicle contains a _____ and the target membrane contains a _____ and the 2 must interact to allow the vesicle to ultimately fuse and release its contents.
nSNARE; tSNARE
_____ allows the interaction bw the vSNARE and the tSNARE to occur, leading to secretion of NT.
calcium
how do botulism toxin and tetanus toxin inhibit NT secretion?
they are proteases (they cleave the SNARE proteins)
which synaptic toxin does black widow spider venom contain?
alpha-latrotoxin
how does alpha-latrotoxin induce a massive non-specific release of NT?
by by-passing the calcium regulation of NT secretion
what do you call complementary proteins found on both the vesicular and target membrane?
SNAREs
which type of SNARE is synaptobrevin?
vSNARE
which type of SNARE is SNAP-25 and syntaxin?
tSNARE
other than vSNARE, what is the other vesicular protein?
synaptotagmin
what does synaptotagmin bind?
calcium
other than tSNAREs, what is the other pre-synaptic plasma membrane protein?
neurexin
what are the 4 steps of docking and fusion of vesicles?
1) vesicle docks
2) SNARE complexes form, pull membranes together
3) entering Ca binds synaptotagmin
4) Ca-bound synaptotagmin catalyzes membrane fusion
what are the 3 major categories of classic NTs?
acetylcholine
amino acid
monoamines
name 3 aa NTs
GABA
glycine
glutamate
how are monoamines synthesized?
by replacing -COOH in aa with another functional group
name the monoamines (2)
catecholamines
indolamines
name the subcategories of catecholamines (3)
norepinephrine
epinephrine
doapmine
name the indolamines (2)
seretonin
histamine
which aa are indolamines formed from?
tryptophan
the subcategory of catecholamines are all modified _____ aa and are closely related to each other
tyrosine
what are the 3 characteristics of classic NTs?
1) small, clear storage vesicles
2) recycled
3) most are synthesized in nerve terminus
T/F non-classic NTs such as peptides are recycled
FALSE. non-classic NTs such as peptides are NOT recycled.
where are non-classic NTs synthesized?
in cell body
4 characteristics of peptide NTs
1) much larger variety
2) may be co-localized to the same nerve terminal as classic NTs
3) most are synthesized in cell body, usually as prohormones
3) most are enzymatically inactivated in the synapse
_____, _____ _____ vesicles often contain “classic” NTs
small, clear round vesicles
_____, _____ vesicles typically contain peptide NTs or, in some cases, _____
large, dense-core; biogenic amines
3 main mechanisms to eliminate NT from cleft
1) reuptake of NT into axon terminus or transport into neighboring glial cell
2) inactivate NT using enzymes present with in synaptic cleft
3) allow NT to diffuse from cleft into surrounding ECF or blood stream
SSRI’s
selective serotonin reuptake inhibitors; antidepressive
Glutamate synthesis and cycling between neurons and glia: The action of glutamate released into the synaptic cleft is terminated by uptake into surrounding glial cells (and neurons) via _____. Within glial cells, glutamate is converted to _____ by _____ and released by glial cells by a different transporter. _____ is taken up into nerve terminals via _____ and converted back to glutamate by _____. Glutamate is then loaded into synaptic vesicles via _____ to complete the cycle.
excitatory amino acid transporters (EAATs)
glutamine
glutamine synthetase
glutamine
SAT2 transporters
glutaminase
vesicular glutamine transporters
acetyl choline esterase (AChE)
cleaves ACh back into choline and acetate
ACh binds to one of several types of _____ receptors on the post-synaptic membrane
cholinergic
AChE inhibitors include _____ (a nerve gas) and many _____
sarin; many organophosphate pesticides
fast chemical synapses contain _____ receptors (_____ channels)
ionotropic; ligand-gated
slow chemical synapses contain _____ receptors (_____ receptors)
metabotropic; G-protein coupled
within fast chemical synapses, there are 2 subcategories based on how quickly a PSP develops in the postsynaptic cell. the speed with which the response is induced is due to differences in the __________.
type of receptor found in the dendritic membrane
how fast are the fast chemical synapses?
~1 msec
how slow are the slow chemical synapses?
secs-mins
in the slow chemical synapses, receptor _____ leads to a change in membrane potential in post-synaptic cell
indirectly
bw fast and slow chemical synapses, which one allows for amplification of the signal, leading to a larger response?
slow chemical synapse
ECF [K+]
4 mM
T/F ions can passively move up their concentration gradient if it brings the membrane potential closer to that ions equilibrium potential
TRUE
the membrane potential of a cell is -20mV, E of Cl- is -45 mV. the net flux of Cl- will be:
inward directed
the axonal membrane domain is non-conducting with regards to Na+ when the voltage-gated Na+ channel is
both closed and inactivated
which vesicular membrane-associated protein is involved in vesicular fusion to the axon terminal membrane?
synaptobrevin
the end-plate potential is the result of which of the following ionic events occurring at the nicotinic AchR?
influx of Na+ and efflux of K+
function of ARP 2/3
nucleation
function of myosin
motor protein
function of CapZ
plus end cap
function of gelsolin
severing
during the cross bridge cycle in a sarcomere, when does the “power stroke” of the myosin head occur?
during the rigor phase
the depolarization phase of an AP is carried by Ca2+ in which cells?
cardiac pacemaker cells and some smooth muscle cells
excitation-contraction coupling refers to:
how a muscle membrane AP leads to an increase in cytoplasmic [Ca2+]
malignant hypothermia can occur when _____ is mutated
RyR1 of skeletal muscle
according to the chemiosmotic hypothesis, which of the following contributes to the proton-motive force across the inner mitochondrial membrane (IMM)?
- the proton concentration gradient across the IMM
- the electrical potential gradient across the IMM
- the difference in oxygen concentration across the IMM
which ionotropic receptor? heteromeric, 5 subunits; number of different subunits varies between muscles and neuronal types; permeable to Na and K
nAChR