INTRODUCTION This application implements the model described in Steephen, J. E. (2009). Excitability range of medium spiny neurons widens through the combined effects of inward rectifying potassium current inactivation and dopaminergic modulation. Neurocomputing, 74.18 (2011): 3884-3897 This help window provides instructions for replicating the results in the paper. The information in this window is available in the text file 'Help.txt' within the 'support_files' folder. If required, it could be printed for easy reference. Launching the program (main.hoc) invokes 4 windows: NEURON Main Menu, Command Window (nrniv), the Control Panel and a Graph Window. Note that the Control Panel is organized into 3 vertical panels. The instructions given below for each figure assume that the application is in its default settings. It is recommended that the application be restarted after each simulation as it would reset all parameters to their default values. Note that you may have to right click the graph panel and select View...>View=plot to view some of the plots. For questions and queries, send e-mail to john.steephen@duk.ac.in. THE SUMMARY FILE In the project folder, you will find a text file named Summary.txt. This file contains the summary of the last completed simulation protocol. This summary file is updated at the end of every simulation. If multiple simulations were conduced during one session, only the summary of last simulation procedure is maintained. Note that one simulation protocol might contain multiple iteration of cell-stimulation, each time varying a setting, as required by the protocol. Thus, one instance of simulation protocol is started when the "START" button is pressed and finished when the message "Summary.txt Updated" is shown in the Command Window. The Summary file is not updated if the user press the "STOP" button to abort the simulation protocol. A copy of the Summary.txt is also created at the end of each simulation. By default, this file is kept in the folder support_files as a file named Output.txt. The name of this backup file can be updated by the user at any point of time, by clicking the "Save to..." button under the RUN section of the control panel. When the simulation gets over, the Summary file is first updated, and a copy will be made on the User-given file. This feature is very useful if a user intends to run multiple simulation procedures one after another. The "Save to.." button is pressed before pressing "START" button to update the output file. The procedure to generate each of the figures shown in the reference paper is given below. Many of the figures are generated with the help of the Summary file. FIGURE 1 Figure one shows the spiking behavior of MSP neuron with different cell properties under intrinsic modulation conditions. The voltage recordings (in mV) are obtained from the cell soma and plotted against time in ms. Fig 1A: (a) In control panel, > Click on Synaptic input under INPUT section > Click on START button The plot obtained in the graph is the required output (b) In control panel, > Click on the inKIR radio button under CELL PROPERTIES > Click Synaptic input statusbutton > Click on the START button The plot is obtained in the graph window. Fig 1B: (a) In control panel, > Click on D1 radiobutton under DA Receptor Variant > Click on radiobutton "Intrinsic" Under DA Modulation Setting > Click on Synaptic input statusbutton > Click on the START button The plot is obtained in the graph (b) In control panel, > Select inKIR radiobutton > Select D1 > Select Intrinsic > Turn on Synaptic input > START simulation Plot is obtained in Graph Fig 1C: (a) Same as 1B(a), except that D2 is chosen instead of D1 (b) Same as 1B(b), except that D2 is chosen instead of D1 FIGURE 2 This figure shows the variation in the no on spikes per up-state with respect to the variations in cell properties and under intrinsic modulation conditions. There are no subfigures, but the six points and the corresponding error bar shown in the figure needs to be obtained using six separate simulations. As an example, the procedure to obtaine the fourth point, labeled inKIR+D1 is given below: In control panel > Select inKIR > Select D1 > Select Intrinsic DA Modulation > Turn on Synaptic input > Set iterations = 30 (under ACTION) > Press "Save to..." button and set output file as Output2.txt > Click START to start the simulation (This will take some time. You may close the Graph panel to speed up the simulatio) When the message "Simulation Complete" appears in the command window, open the file Output1.txt. In that file under ACTION and RESULTS section, the mean and the s.e.m of the No. of spikes per Up-state is provided. (Took me 5.5 mins to finish simulation and the values obtained are Mean = 3.2 and s.e.m = 0.1) These are the values plotted as fourth point and its error bar in Fig. 2. Similarly the rest of the points can be generated. FIGURE 3 This is a figure derived from Fig. 2 described above. FIGURE 4 Fig 4A: Spike frequency vs Current Injection amplitude under intrinsic dopamine modulation conditions. For each line shown in the figure, one separate simulation is needed. As an example, describing the procedure to produce the plot curresponding to non-inKIR D2. In control panel, > Select D2 > Select Intrinsic > Select 'Range Run' radiobutton under ACTION > set min = 0.21 > set max = 0.3 > set iterations = 19 > Press "Save to..." button and set output file as Output4a.txt > Click START to start the simulation When "Simulation Complete" message appear in the command window, open the file Output4a.txt. You can find a table showing the IC amplitude and the corresponding spike frequency. This can be plotted using plotly, python, MS Excel, Origin, or whatever plotting software you are comfortable with. The other lines in Fig. 4A can be produced by varying the cell properties, but maintaining the settings of the INPUT section and the ACTION section unchanged. Fig 4B: Spike frequency vs input Up-state frequency under intrinsic dopamine modulation conditions. This is similar to the procedure in Fig. 4A, but there are some changes in INPUT and ACTION section settings. Describing the procedure to produce plot corresponding to non-inKIR D2. Marking (*) wherever there are changes from the procedure in Fig. 4A above In control panel, > Select D2 > Select Intrinsic > (*) Turn on Synaptic input > Select 'Range Run' > (*) set min = 5 > (*) set max = 8.5 > (*) set iterations = 15 > Press "Save to..." button and set output file as Output4b.txt > Click START (long simulation: close the graph for better speed) When "Simulation Complete", open the Output4b.txt file, where you will find the table showing the variation of spike frequency vs the input up-state frequency. Use a convenient plotting tool to visualize the plot. FIGURE 5 This figure is generated using a procedure exactly identical to the Fig. 2. After generating the Output file through the simulation, open the file and observe the mean and s.e.m. of the Spike Onset Time given in the ACTION and RESULTS. This value represent one point in the Fig. 5, corresponding to the Cell Properties mentioned in the simulation. The process can be repeated for different cell properties and the rest of the points can be plotted. If the output files used to obtain Fig. 2 is available, there is no need to redo the simulation to obtain Fig. 5 FIGURE 6 This figure is generated from the data of Fig. 5. No separate dimulation needed. Thus the series of simulations conducted to generate Fig. 2 can produce data for preparing Figs 3, 5, and 6 as well. FIGURE 7 Identical procedure as in Fig. 1 - except one setting: > Intrinsic-synaptic option is selected for dopamine modulation FIGURE 8 Identical procedure as in Fig. 2 - except one setting: > Intrinsic-synaptic option is selected for dopamine modulation FIGURE 9 Data from Fig. 8 is used to prepare this figure. FIGURE 10 Identical procedure as in Fig. 4 - except one setting: > Intrinsic-synaptic option is selected for dopamine modulation FIGURE 11 Identical procedure as in Fig. 5 - except one setting: > Intrinsic-synaptic option is selected for dopamine modulation