%plotting 3rewards data clear all; load ('R_3R.mat'); load ('RAW.mat'); %get parameters BinBase=R_3R.Param.BinBase; BinDura=R_3R.Param.BinDura; bins=R_3R.Param.bins; binint=R_3R.Param.binint; binstart=R_3R.Param.binstart; PStatBins=0.01; %using more stringent cutoff to reduce pre-delivery noise %which bins bound the area examined for reward-selectivity? (in seconds) Time1=0.4; %seconds Time2=3; %seconds Bin1=(((Time1-BinDura(2)/2)-binstart)/binint); %convert to bin name Bin2=(((Time2-BinDura(2)/2)-binstart)/binint); %convert to bin name %sorting bin -- which bin the neurons' activity is sorted on for heatmap(in seconds) SortBinTime=1.1; %seconds SortBin=round((((SortBinTime-BinDura(2)/2)-binstart)/binint)); %convert to bin name sucrose=[1 0.6 0.1]; maltodextrin=[.9 0.3 .9]; water=[0.00 0.75 0.75]; total=[0.3 0.1 0.8]; exc=[0 113/255 188/255]; inh=[240/255 0 50/255]; %% conduct lick analysis global Dura Tm BSIZE Tbin path='C:\Users\dottenh2\Documents\MATLAB\David\2Rewards Nex Files\3RLick_paper.xls'; %Main settings BSIZE=0.01; %Do not change Dura=[-22 20]; Tm=Dura(1):BSIZE:Dura(2); Tbin=-0.5:0.005:0.5; %window used to determine the optimal binsize MinNumTrials=5; Lick=[];Lick.Ninfo={};LL=0;Nlick=0; %Smoothing Smoothing=1; %0 for raw and 1 for smoothing SmoothTYPE='lowess'; %can change this between lowess and rlowess (more robust, ignores outliers more) SmoothSPAN=50; %percentage of total data points if Smoothing~=1, SmoothTYPE='NoSmoothing';SmoothSPAN=NaN; end % List of events to analyze and analysis windows EXTRACTED from excel file [~,Erefnames]=xlsread(path,'Windows','a3:a8'); % cell that contains the event names %Finds the total number of sessions for i=1:length(RAW) if strcmp('TH',RAW(i).Type(1:2)) Nlick=Nlick+1; Lick.Linfo(i,1)=RAW(i).Ninfo(1,1); end end Lick.Erefnames= Erefnames; %preallocating the result matrix for k=1:length(Erefnames) Lick.Ev(k).PSTHraw(1:Nlick,1:length(Tm))=NaN(Nlick,length(Tm)); Lick.Ev(k).BW(1:Nlick,1)=NaN; Lick.Ev(k).NumberTrials(1:Nlick,1)=NaN; end for i=1:length(RAW) %loops through sessions if strcmp('TH',RAW(i).Type(1:2)) LL=LL+1; %lick session counter for k=1:length(Erefnames) %loops thorough the events EvInd=strcmp(Erefnames(k),RAW(i).Einfo(:,2)); %find the event id number from RAW LickInd=strcmp('Licks',RAW(i).Einfo(:,2)); %find the event id number from RAW if sum(EvInd)==0 fprintf('HOWDY, CANT FIND EVENTS FOR ''%s''\n',Erefnames{k}); end Lick.Ev(k).NumberTrials(LL,1)=length(RAW(i).Erast{EvInd}); if ~isempty(EvInd) && Lick.Ev(k).NumberTrials(LL,1)>MinNumTrials %avoid analyzing sessions where that do not have enough trials [PSR1,N1]=MakePSR04(RAW(i).Erast(LickInd),RAW(i).Erast{EvInd},Dura,{1});% makes collpased rasters. PSR1 is a cell(neurons) if ~isempty(PSR1{1}) %to avoid errors, added on 12/28 2011 %forcing 100ms bin size to keep it consistent across %sessions (no reason is should be different for licks) [PTH1,BW1,~]=MakePTH07(PSR1,repmat(N1, size(RAW(i).Erast{LickInd},1),1),{2,0,0.1});%these values force bin size to be 100ms PTH1=smooth(PTH1,SmoothSPAN,SmoothTYPE)'; %------------- Fills the R.Ev(k) fields -------------- Lick.Ev(k).BW(LL,1)=BW1; Lick.Ev(k).PSTHraw(LL,1:length(Tm))=PTH1; end end end end end Xaxis=[-2 12]; Ishow=find(Tm>=Xaxis(1) & Tm<=Xaxis(2)); time1=Tm(Ishow); c=[-1000 7500];ClimE=sign(c).*abs(c).^(1/4);%ColorMapExc colormap(jet); sucrose=[1 0.6 0.1]; maltodextrin=[.9 0.3 .9]; water=[0.00 0.75 0.75]; total=[0.3 0.1 0.8]; exc=[0 113/255 188/255]; inh=[240/255 0 50/255]; Ev1=strcmp('RD1', Lick.Erefnames); Ev2=strcmp('RD2', Lick.Erefnames); Ev3=strcmp('RD3', Lick.Erefnames); psth1=nanmean(Lick.Ev(Ev1).PSTHraw(:,Ishow),1); sem1=nanste(Lick.Ev(Ev1).PSTHraw(:,Ishow),1); %calculate standard error of the mean up1=psth1+sem1; down1=psth1-sem1; psthE=nanmean(Lick.Ev(Ev2).PSTHraw(:,Ishow),1); semE=nanste(Lick.Ev(Ev2).PSTHraw(:,Ishow),1); %calculate standard error of the mean upE=psthE+semE; downE=psthE-semE; psth3=nanmean(Lick.Ev(Ev3).PSTHraw(:,Ishow),1); sem3=nanste(Lick.Ev(Ev3).PSTHraw(:,Ishow),1); %calculate standard error of the mean up3=psth3+sem3; down3=psth3-sem3; %plotting subplot(2,3,1); hold on; plot(time1,psth1,'Color',sucrose,'linewidth',1); plot(time1,psthE,'Color',maltodextrin,'linewidth',1); plot(time1,psth3,'Color',water,'linewidth',1); patch([time1,time1(end:-1:1)],[up1,down1(end:-1:1)],sucrose,'EdgeColor','none');alpha(0.5); patch([time1,time1(end:-1:1)],[upE,downE(end:-1:1)],maltodextrin,'EdgeColor','none');alpha(0.5); patch([time1,time1(end:-1:1)],[up3,down3(end:-1:1)],water,'EdgeColor','none');alpha(0.5); title('Mean lick rate'); %plot([-2 10],[0 0],':','color','k'); plot([0 0],[-2 8],':','color','k','linewidth',0.75); axis([-2 12 0 7.5]); xlabel('Seconds from reward delivery'); ylabel('Licks/s'); legend('Sucrose trials','Maltodextrin trials','Water trials'); %% bins analysis %first and last bin aren't included because you can't compare to the previous/subsequent bin %this axis plots the bins on their centers xaxis=linspace(binstart+binint+BinDura(2)/2,binstart+(bins-2)*binint+BinDura(2)/2,bins-2); for i=2:(bins-1) %no including first or last bin because can't compare to previous/subsequent bin %finds out whether firing is stronger (high excitation or lower inhibition) for 1 or 2 for k=1:length(R_3R.Ninfo) %runs through neurons if R_3R.BinStatRwrd{i,1}(k).IntSig < PStatBins %if neuron is significant for this bin if R_3R.BinStatRwrd{i-1,1}(k).IntSig < PStatBins || R_3R.BinStatRwrd{i+1,1}(k).IntSig < PStatBins %either previous or subsequent bin must be significant if R_3R.BinStatRwrd{i,1}(k).R1Mean >= R_3R.BinStatRwrd{i,1}(k).R2Mean && R_3R.BinStatRwrd{i,1}(k).R1Mean >= R_3R.BinStatRwrd{i,1}(k).R3Mean %if firing is greater on sucrose trials than maltodextrin and water. if there's a tie, make it sucrose (this is highly unlikely) R_3R.BinRewPref{i,1}(k,1)=1; %sucrose preferring elseif R_3R.BinStatRwrd{i,1}(k).R2Mean > R_3R.BinStatRwrd{i,1}(k).R1Mean && R_3R.BinStatRwrd{i,1}(k).R2Mean >= R_3R.BinStatRwrd{i,1}(k).R3Mean %if mal is greater than suc and water R_3R.BinRewPref{i,1}(k,1)=2; %maltodextrin preferring else R_3R.BinRewPref{i,1}(k,1)=3; %water preferring end else R_3R.BinRewPref{i,1}(k,1)=0; %if not significant in 2 consecutive bins end else R_3R.BinRewPref{i,1}(k,1)=0; %if no sig reward modulation end end %find how many NAc neurons have significant reward modulation in each bin NN1perBin(i,1)=sum(R_3R.BinRewPref{i,1}(:,1)==1); %sucrose pref NN2perBin(i,1)=sum(R_3R.BinRewPref{i,1}(:,1)==2); %malto pref NN3perBin(i,1)=sum(R_3R.BinRewPref{i,1}(:,1)==3); %malto pref NNperBin(i,1)=sum(R_3R.BinRewPref{i,1}(:,1)>0); %any %normalize to number of neurons in population NN1norm=NN1perBin./length(R_3R.Ninfo); NN2norm=NN2perBin./length(R_3R.Ninfo); NN3norm=NN3perBin./length(R_3R.Ninfo); NNnorm=NNperBin./length(R_3R.Ninfo); end %plotting number of significantly modulated neurons across time %NAc subplot(2,3,2); hold on; plot(xaxis,NNnorm(2:bins-1),'Color', total,'linewidth',1.5); plot(xaxis,NN1norm(2:bins-1),'Color',sucrose,'linewidth',1.5); plot(xaxis,NN2norm(2:bins-1),'Color',maltodextrin,'linewidth',1.5); plot(xaxis,NN3norm(2:bins-1),'Color',water,'linewidth',1.5); plot([Time1 Time1],[-1 1],':','color','k','linewidth',0.75); plot([Time2 Time2],[-1 1],':','color','k','linewidth',0.75); axis([xaxis(1) xaxis(end) 0 0.85]); legend('Total','Suc > mal & wat','Mal > suc & wat','Water > suc & mal','Location','northeast') ylabel('Fraction of population'); xlabel('Seconds from RD'); title('Reward-selective neurons'); %% plotting sucrose-selective neurons %color map [magma,inferno,plasma,viridis]=colormaps; colormap(plasma); c=[-100 2000];ClimE=sign(c).*abs(c).^(1/4);%colormap %events we're looking at RD1=strcmp('RD1', R_3R.Erefnames); RD2=strcmp('RD2', R_3R.Erefnames); RD3=strcmp('RD3', R_3R.Erefnames); %setting up parameters Xaxis=[-2 5]; inttime=find(R_3R.Param.Tm>=Xaxis(1) & R_3R.Param.Tm<=Xaxis(2)); paramtime=R_3R.Param.Tm(inttime); %find all neurons with greater firing for sucrose for i = 1:(Bin2-Bin1+1) %the added +1 is necessary because bin 20 is the 21st entry in the matrix Pref1(:,i)=R_3R.BinRewPref{Bin1+i}==1; %get neurons that have greater firing for sucrose in any of the bins bounded above Resp11(:,i)=Pref1(:,i)&cat(1,R_3R.BinStatRwrd{Bin1+i,1}.SucRespDir)==1; %get neurons with excitation to sucrose Resp12(:,i)=Pref1(:,i)&cat(1,R_3R.BinStatRwrd{Bin1+i,1}.MalRespDir)==1;%get neurons with inhibition to maltodextrin Resp13(:,i)=Pref1(:,i)&cat(1,R_3R.BinStatRwrd{Bin1+i,1}.WatRespDir)==-1;%get neurons with inhibition to maltodextrin end Sel=sum(Pref1,2)>0; %all neurons selective in any bin Sel1=sum(Resp11,2)>0; %all selective neurons sucrose excited in any bin Sel3=sum(Resp12,2)>0; %all selective neurons malto inhibited in any bin Sel6=sum(Resp13,2)>0; %all selective neurons malto inhibited in any bin subplot(2,4,5); %heatmap of suc preferring neurons' response to sucrose Neurons=R_3R.Ev(RD1).PSTHz(Sel,inttime); %get the firing rates of neurons of interest SucResp=cat(1,R_3R.BinStatRwrd{SortBin+1,1}.R1Mean); %sucrose responses TMP=SucResp(Sel); %find the magnitude of the excitations for water for this binTMP(isnan(TMP))=0; %To place the neurons with no onset/duration/peak at the top of the color-coded map [~,SORTimg]=sort(TMP); Neurons=Neurons(SORTimg,:); %sort the neurons by magnitude imagesc(paramtime,[1,sum(Sel,1)],Neurons,ClimE); title(['Sucrose trials']); xlabel('Seconds from RD'); ylabel('Individual neurons'); hold on; plot([0 0],[0 sum(Sel)],':','color','k','linewidth',0.75); subplot(2,4,6); %heatmap of suc preferring neurons' response to maltodextrin Neurons=R_3R.Ev(RD2).PSTHz(Sel,inttime); %get the firing rates of neurons of interest Neurons=Neurons(SORTimg,:); %sort the neurons same as before imagesc(paramtime,[1,sum(Sel,1)],Neurons,ClimE); title(['Maltodextrin trials']); xlabel('Seconds from RD'); hold on; plot([0 0],[0 sum(Sel)],':','color','k','linewidth',0.75); subplot(2,4,7); %heatmap of suc preferring neurons' response to water Neurons=R_3R.Ev(RD3).PSTHz(Sel,inttime); %get the firing rates of neurons of interest Neurons=Neurons(SORTimg,:); %sort the neurons same as before imagesc(paramtime,[1,sum(Sel,1)],Neurons,ClimE); title(['Water trials']); xlabel('Seconds from RD'); hold on; plot([0 0],[0 sum(Sel)],':','color','k','linewidth',0.75); %plot suc preferring neurons to suc psthE=nanmean(R_3R.Ev(RD1).PSTHz(Sel,inttime),1); semE=nanste(R_3R.Ev(RD1).PSTHz(Sel,inttime),1); %calculate standard error of the mean upE=psthE+semE; downE=psthE-semE; %plot suc preferring neurons to malt psth2=nanmean(R_3R.Ev(RD2).PSTHz(Sel,inttime),1); sem2=nanste(R_3R.Ev(RD2).PSTHz(Sel,inttime),1); %calculate standard error of the mean up2=psth2+sem2; down2=psth2-sem2; %plot suc preferring neurons to water psth3=nanmean(R_3R.Ev(RD3).PSTHz(Sel,inttime),1); sem3=nanste(R_3R.Ev(RD3).PSTHz(Sel,inttime),1); %calculate standard error of the mean up3=psth3+sem3; down3=psth3-sem3; %plotting subplot(2,3,3); hold on; plot(paramtime,psthE,'Color',sucrose,'linewidth',1); plot(paramtime,psth2,'Color',maltodextrin,'linewidth',1); plot(paramtime,psth3,'Color',water,'linewidth',1); patch([paramtime,paramtime(end:-1:1)],[upE,downE(end:-1:1)],sucrose,'EdgeColor','none');alpha(0.5); patch([paramtime,paramtime(end:-1:1)],[up2,down2(end:-1:1)],maltodextrin,'EdgeColor','none');alpha(0.5); patch([paramtime,paramtime(end:-1:1)],[up3,down3(end:-1:1)],water,'EdgeColor','none');alpha(0.5); legend('Suc trials','Mal trials','Wat trials'); plot([-2 5],[0 0],':','color','k','linewidth',0.75); plot([0 0],[-2 8],':','color','k','linewidth',0.75); axis([-2 5 -2 4.7]); ylabel('Mean firing (z-score)'); title(['Suc>mal&wat (n=' num2str(sum(Sel)) ' of ' num2str(length(Sel)) ')']) xlabel('Seconds from RD'); G = sum(Sel1&Sel3&Sel6); F = sum(Sel3&Sel6); E = sum(Sel1&Sel6); D = sum(Sel1&Sel3); A = sum(Sel1); B = sum(Sel3); C = sum(Sel6); %vertical venn diagram x = [0 0 1 1]; y1 = [C-E C-E+A C-E+A C-E]; y2 = [C-F C-F+B C-F+B C-F]; y3 = [0 C C 0]; subplot(2,35,64); hold on; s = patch(x,y1,sucrose); m = patch(x,y2,maltodextrin); w = patch(x,y3,water); alpha(s,0.7); alpha(w,0.5); alpha(m,0.5); set(gca,'xtick',[]); ylabel('Distribution of neurons'); axis([0 1 0 C-E+A]); %% stats on reward firing averaged together %for simplicity, just looking at average activity in sort bin %because I already have that data collected SucResp=cat(1,R_3R.BinStatRwrd{SortBin+1,1}.R1Mean); %suc responses MalResp=cat(1,R_3R.BinStatRwrd{SortBin+1,1}.R2Mean); %mal responses WatResp=cat(1,R_3R.BinStatRwrd{SortBin+1,1}.R3Mean); %wat responses [~,R_3R.RewRespStat{1,1},R_3R.RewRespStat{1,2}]=anovan(cat(1,SucResp(Sel),MalResp(Sel),WatResp(Sel)),{cat(1,zeros(sum(Sel),1),ones(sum(Sel),1),2*ones(sum(Sel),1)),cat(1,R_3R.Ninfo(Sel,4),R_3R.Ninfo(Sel,4),R_3R.Ninfo(Sel,4))},'varnames',{'Reward','Subject'},'random',2,'Display','off','model','full'); R_3R.RewRespStat{1,3}=multcompare(R_3R.RewRespStat{1,2},'display','off'); save('R_3R.mat','R_3R');