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1 Bi/CNS/NB 150 Problem Set 3 Due: Tuesday, Oct. 27, at 4:30 pm Instructions: 1) Drop off in the Bi 150 box outside Baxter 331 or to the head TA (jcolas). 2) Submit with this cover page. 3) Use a separate sheet of paper for each problem. 4) Type all answers if possible. 5) Use complete, grammatically correct sentences. 6) Include your name and the page number on every page. 7) Note that late problem sets receive a 10% deduction for every day past the due date. Name: Time and date submitted: Total pages (including cover page): Comments: Problem 1 grade: Problem 1 comments: Problem 2 grade: Problem 2 comments: Total grade: 1

2 Problem 1 (1.5 points): Synaptic integration Problem 1.A (0.9 points): Inhibition Many inhibitory synapses in the CNS utilize GABA A and glycine receptors. Both GABA A and glycine receptor channels are permeable to Cl - ions. 1.A.a. Calculate the Nernst potential for Cl - ions, E Cl, at 25 C. Consult the ionic concentrations listed on the fourth slide from the second half of the first lecture. 1.A.b. The resting membrane potential of neurons is sometimes close to E Cl. When this is the case, how does the membrane voltage change if only GABA A receptors are activated? 1.A.c. Consider a neuron receiving two excitatory inputs as shown below. The traces at the right show the membrane potential in the soma without inhibition. i) Excitatory input 1 is stimulated alone. ii) Excitatory inputs 1 and 2 are stimulated simultaneously. Excitatory input 1 Excitatory input 2 dendrite A Inhibitory input i) ii) B 1 alone 1+2 soma time For each trace, redraw the trace and then superimpose the trace if a GABAergic input were active at location A during the excitatory stimulation. Repeat for location B. Explain the reasoning behind each trace. 2

3 Problem 1.B (0.6 points): Active dendrites 1.B.a. Panel B in the figure below shows simultaneous recordings of the membrane potential from the soma and a dendrite when an action potential is induced by injection of current into the soma. Panel A shows the recording sites. A B Soma Dendrite time Redraw the dendritic traces and then superimpose the dendritic voltage trace during action potentials in a mutant animal that lacks voltage-gated Na + channels at the dendrites. The action potentials are induced in the same way as before by injecting current at the soma. Assume that the only difference is in the dendrites and their lack of voltage-gated Na + channels. Explain the reasoning behind the traces. 1.B.b. In the case of a normal neuron, how might backpropagating spikes due to dendritic Na + channels influence the coincidence-detection properties of NMDA receptors? 1.B.c. Describe the roles played by Na +, Ca 2+, and Mg 2+ ions during coincidence detection. 3

4 Problem 2 (1.5 points): Synaptic transmission Problem 2.A (0.3 points): Chronology Write the following events in the correct sequence as they occur at the neuromuscular junction of the California olive-tree rat (Rattus pasadensis) during normal synaptic transmission. If an event overlaps in time with any of the others, explain this in your answer. Begin from event 9. 1) Action potential arrives at the presynaptic terminal. 2) Muscle fiber action potential. 3) Na + enters, and K + exits through the same channels. 4) There is a local increase in intracellular Ca 2+ concentration near the Ca 2+ channels. 5) ACh is hydrolyzed to acetate and choline. 6) Choline is transported into the presynaptic terminal. 7) ACh molecules fill the synaptic vesicles. 8) ACh is released into the synaptic cleft. 9) The synaptic vesicles become acidic. (Begin here.) 10) Na + is pumped out, and K + pumped into the presynaptic terminal and muscle fiber. 11) ACh binds to ACh-gated ion channels on the postsynaptic membrane. 12) ACh diffuses within the synaptic cleft. 13) Muscle fiber reaches threshold depolarization. 14) ACh-gated ion channels open. 15) ACh-filled vesicles fuse with the presynaptic membrane. 16) Ca 2+ enters the presynaptic terminal through voltage gated Ca 2+ channels. 4

5 Problem 2.B (1.2 points): Excitatory postsynaptic potentials You are in the lab recording from neuron pairs each connected by a glutamatergic synapse in four different mutant animals and in a wild-type animal. In each animal a different protein involved in neurotransmitter release is mutated. You stimulate the presynaptic cell and record from the postsynaptic neuron of each pair. For each mutation below, describe the following: i) After the first stimulus, how does the excitatory postsynaptic potential (EPSP) you record from the postsynaptic neuron differ from the EPSP recorded from a wild-type animal? Does it increase, decrease, stay comparatively the same, or fail to be generated at all? ii) After five stimulations of the presynaptic cell in quick succession, how does the EPSP you record from the mutant animal differ from that in the same experiment on a wild-type animal? Explain your reasoning in each case. The mutations are as follows: 2.B.a. Mutation of the voltage-gated Ca 2+ channels of the presynaptic terminal. The mutant channels have a lower opening threshold. 2.B.b. Mutation of the Ca 2+ binding sites of synaptotagmin. The mutant protein does not bind Ca B.c. Mutation of the glutamate transporter to inhibit its function. Several redundant processes maintain the intracellular glutamate concentration of ~10 mm. Contrast this with the concentration of ~1 μm in cerebrospinal fluid. 2.B.d. Knockout of N-ethylmaleimide-sensitive factor (NSF). 5

1) Drop off in the Bi 150 box outside Baxter 331 or to the head TA (jcolas).

1) Drop off in the Bi 150 box outside Baxter 331 or  to the head TA (jcolas). Bi/CNS/NB 150 Problem Set 3 Due: Tuesday, Oct. 27, at 4:30 pm Instructions: 1) Drop off in the Bi 150 box outside Baxter 331 or e-mail to the head TA (jcolas). 2) Submit with this cover page. 3) Use a

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