Bio 304 Physiology of Neural Systems 2008 Tutorial Assignment # 9 Student name:______________________ T.A.

.______________________________ Tutorial: Site of Impulse Initiation Tutorial Read the general instructions for completing NIA assignments (available from the 304 home webpage) before starting this tutorial. Part I. Where is the site of spike initiation? Follow instructions outlined in the tutorial for this procedure. Do not adjust the speed button in the Run Control panel Important difference in this tutorial from lecture: the soma in the first part of this simulation possesses voltageactivated channels and can create its own propagated AP. If exact values do not appear on the Run Control time (ms) field, close and re-open simulation. View the Vm from the dendrite to the axon in the Voltage vs Space panel using Reset then "Continue for (ms)" Use these electrode location distances (assuming 0 um = the point where the dendrites join the soma) exactly as plotted on the Voltage vs Space Graph for all of Part I: 1. Dendrite: -750 um 2. Soma: 100 um 3. Axon: 888.776 um Using an AlphaSynapse[0] Gmax of 15 uS and with the Voltage vs Space graph report the Vmax (all one dpa) at the electrode locations for the following times after PSP onset (use the Continue for (ms) to freeze the delta Vm for these measurements). 0.1 each = (4.2 marks) Time after PSP onset (ms) 0.25 0.50 0.75 1.05 1.25 1.50 1.75 2.05 2.25 2.50 2.75 3.05 3.25 3.50 Vmax (dendrite electrode) (mV) -53.4 -51.4 -51.2 -50.9 -50.4 -49.7 -48.8 -47.0 -44.7 -39.8 -32.5 -20.4 -13.1 -9.5 Vmax at Vmax at axon soma (electrode #2) (electrode #3) (mV) (mV) -54.6 -63.3 -51.5 -60.1 -51.0 -57.8 -50.5 -55.5 -50.0 -53.8 -49.2 -50.8 -48.1 -45.1 -46.0 -21.8 -43.2 18.8 -37.6 26.1 -29.5 9.4 -16.6 -11.8 -10.0 -24.1 -8.3 -37.7 Student number (only last 4 digits):_______________ Date:_____________________________

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Student number (last 4 digits):__________________

Attach a NIA graph showing the Voltage vs Space graph 2.4 ms following PSP onset. (2 marks)

What difference is there in the AP Vmax as recorded at the dendrite compared to at the soma? Explain your answer? (3 marks) The AP is reduced at the dendrite because this part of the neuron possesses no voltage-gated channels. It will therefore exhibit only a passive delta Vm shift which will be reduced because of the intracellular resistance encountered by the current actively generated by the soma AP. Use the AlphaSynapse [0] panel to move the site of the synapse and Adjust AlphaSynapse reversal voltage to -10 mV. Using the Voltage vs Time graph and a synaptic strength of Gmax = 15 uS report the time (from PSP onset) to AP Vmax at the electrode locations for the following synapse positions (manually adjust the View settings to measure Vm at the soma and dendrite electrodes). Then determine the minimum synaptic strength to evoke the axon AP for those synaptic positions. (use one dpa for all values) 0.1 each = (4.4 marks) Position of synapse (um) (slide bar setting) -1500 (1.000) -1410 (0.944) -1250 (0.833) -1083 (0.722) -917 (0.611) -750 (0.500) -584 (0.389) -417 (0.278) -251 (0.167) -84 (0.056) 0 (0.000) Time of Vmax at dendrite (ms) 3.1 3.0 2.9 2.8 2.8 2.7 2.7 2.7 2.7 2.7 2.7 Time of max Vmax at soma (ms) 3.0 2.9 2.8 2.8 2.7 2.7 2.6 2.6 2.6 2.6 2.6 Time of max Vmax at axon (ms) 2.2 2.2 2.1 2.1 2.1 2.0 2.0 2.0 2.0 2.0 2.0 Synaptic strength for AP threshold (uS)) 12.0 11.9 11.8 11.7 11.6 11.5 11.5 11.4 11.4 11.4 11.4

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Student number (last 4 digits):__________________

Does the time taken to reach AP Vmax change with the position of the synapse? Explain your answer. (3 marks) The time to AP Vmax reduces as the synapse approaches the spike-initiation site because the total capacitance that the synaptic current encounters is reduced and the time take to fully charge the membrane reduces therefore activating an axon AP sooner. i.e Passive decay is reduced. What will be the effect of making the synapse reverse potential more negative to the time take to reach AP Vmax? To conductance required for achieving axon threshold? Explain. (4 marks) As Erev becomes more negative both the time taken to achieve AP Vmax, and the conductance required to achieve threshold will increase. Erev defines the maximum voltage a membrane can achieve due to a synapse. As Erev becomes more negative, the synapse would need to be open longer, and have higher conductance in order to achieve the same threshold voltage thus the longer delay, and higher conductance required. In lecture we discussed the example of a neuron where there was no time delay in the appearance of an AP at the spike-initiation site and at the soma. Why is there a time delay in this simulation? (3 marks) The lecture discussed the case of a neuron that had no voltage-gated channels in the soma and would not propagate an AP. The current created from the AP generated at the spike-initiation site would therefore immediately create a delta Vm at the soma. In this simulation the soma possesses channels and would therefore propagate an AP with a conduction velocity thereby resulting in a time delay. Why does the synaptic strength (i.e., synaptic conductance) to achieve AP threshold change as the synapse approaches the soma? Explain your reasoning. (3 marks) As the synapse approaches the soma site the current created by the synapse encounters less total resistance and therefore results in a higher delta Vm at the AP initiation site. This means it will require less neurotransmitter to result in a postsynaptic AP. Do the axon AP and the soma AP have the same synaptic strength thresholds? Explain your answer. (3 marks) The axon AP possess a lower threshold than the soma AP and fires first because much of the AP current that would fire the soma is lost into the dendrites reducing the soma Vmax and the time taken before the soma channels begin its AP places them into the hyperpolarization phase of the current arriving from the axon AP. Attach a single NIA Voltage vs Time graph with the synapse at default strength and at the following cell locations: -1500 um, -750 um and 0 um. Use the Keep Lines options to show all the lines on one graph. (2 marks)

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Student number (last 4 digits):__________________

Return the position of the synapse to the mid-way point (-750 um from the soma, 0.500 on the sliding bar scale) Reset all values to default. For the following synaptic strengths first use the Voltage vs Space graph to report the PSP Vmax at the dendrite electrode at 0.4 ms following PSP onset. Then, allow the simulation to run for 6 ms and use the Voltage vs Time graph to report the PSP Vmax and with unrounded values, the time from PSP onset to AP Vmax at the axon electrode (one dpa for all values). 0.1 each = (3.6 marks) Synaptic strength Gmax (uS) 13.5 14.5 15.5 16.5 17.5 18.5 19.5 20.5 25.5 30.5 35.5 40.5 PSP Vmax (dendrite) (mV) -52.8 -52.0 -51.3 -50.6 -49.9 -49.2 -48.5 -47.8 -44.7 -41.9 -39.4 -37.1 AP Vmax (axon) (mV) 24.9 28.2 29.8 30.7 31.4 32.0 32.3 32.6 33.5 33.9 34.1 34.3 Time from PSP to AP Vmax (axon) (ms) 3.1 2.6 2.3 2.1 2.0 1.9 1.8 1.7 1.5 1.4 1.3 1.2

Attach a NIA graph from the programme showing the traces from the three recording electrodes at the synaptic strengths used (scale the y axis so that no line runs off the graph by right-clicking on the graph and choosing the View=Plot; show all lines with the Keep Lines option). (2 marks)

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Student number (last 4 digits):__________________

Attach three fully labeled graphs plotting synaptic strength (X-axis) versus: A. Vm at 0.4 ms after synaptic event, B. Axon AP Vmax and, C. Time from synaptic event to reach AP Vmax, (all Y-axis). (6 marks)

Does the amplitude of the depolarization recorded at the dendrite change linearly? Explain your answer. (3 marks) The depolarization increases to a point but levels off as it approaches Erev (-15 mV).

How does the axon AP Vmax change as the synaptic strength increases? Why? (3 marks) The axon AP Vmax increases as synaptic strength increase. This is because the AP occurs sooner and sooner eventually riding on top of the synaptic post-synaptic potential. How does the timing of the axon AP change as the synaptic strength increases? Why? (3 marks) As synaptic strength increases the axon AP occurs sooner because the rate at which the spike initiating site membrane is depolarized increases.

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Student number (last 4 digits):__________________

Part II. How is impulse initiation influenced by cell morphology? Return all settings to default by clicking off all checked buttons Restore the default synaptic location in the synaptic panel to midway along the dendrite (0.500 on the sliding bar scale) for all the following exercises. IN the Cell Parameters panel make the soma passive by eliminating its voltage-gated channels (enter zero in the Na chan density and K chan density buttons of the Active Soma Parameters panel). Report the synaptic strengths (one dpa) required to achieve threshold for the axon AP for the following soma diameters. 0.2 each = (2 marks) Soma diameter (um) 55 105 155 205 255 305 355 455 505 1005 Synaptic strength for AP threshold (uS) 14.6 14.9 15.2 15.6 15.9 16.3 16.6 17.3 17.7 21.2

How does the threshold of the axon AP change as soma diameter increases? Why? (3 marks) As the soma diameter increases the threshold increases because the soma dissipates the current created by the PSP before it has a chance to shift the Vm at the spike-initiating site. Is there a delta Vm still seen at the soma even though it does not possess voltage-gated channels? Explain your answer. (3 marks) Yes, there is still a delta Vm shift seen at the soma and this is caused by the combined passive effects of the currents created by the PSP and the axon AP. Keep the soma Na and K channel density at zero Return the soma diameter and length to default (200 um) Set Total #(ms) to 10 Use the Axon Parameters panel to adjust Na channel density

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Student number (last 4 digits):__________________

Report the synaptic strengths (one dpa) required to achieve threshold for the axon AP for the following axonal Na+ and K+ channel densities. 0.2 each = (2 marks) Axon Na chan density (S/cm2) 0.10 0.12 0.13 0.14 0.15 0.16 0.17 0.18 0.19 0.20 Axon K chan density (S/cm2) 0.038 0.038 0.038 0.038 0.038 0.038 0.038 0.038 0.038 0.038 Synaptic strength for AP threshold (uS) 19.2 16.2 15.0 13.9 12.8 11.8 10.8 9.8 8.8 7.7

How does the threshold of the axon AP change as Na channel density increases? Why? (3 marks) As Na channel density increases the threshold decreases because the Hodgkin cycle is more likely to be activated with more Na channels. For the following exercise keep the soma Na and K channel density at zero Return the axon Na and K channel density to default by clicking the checked buttons Set Total # (ms) to 10 Report the synaptic strengths required to achieve threshold for the axon AP for the following axon diameters. (one dpa for all values) 0.2 each = (2 marks) Axon diameter (um) 20.5 15.5 14.5 13.5 10.5 5.5 4.5 3.5 2.5 1.5 Synaptic strength for AP threshold (uS) 15.8 15.6 15.5 15.5 15.3 15.1 15.0 15.0 14.9 14.6

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Student number (last 4 digits):__________________

How does the synaptic strength to achieve axon AP threshold change as axon diameter decreases? Why? (3 marks) As the axon diameter decreases it requires less synaptic strength to achieve axon AP threshold since the axons internal resistance increases forcing more current to leave via the membrane and increasing the Vm depolarization at the spike initiation site.

What is the disadvantage of decreasing axon diameter to change AP threshold (hint: watch the Voltage vs Space graph as you activate the synaptic event). What can the cell do to its morphology to minimize this disadvantage? (6 marks)

As the axon diameter decreases, the AP conduction velocity decreases slowing the transmission of the AP. The cell can minimize this effect by restricting its axon diameter decrease to only the spike-initiating site. It could also include myelination to the rest of its axon to compensate for the reduced CV at the spike initiation site.

TOTAL ASSIGNMENT MARK: ____ / 76.2