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1 upplementry Informtion Activity Recll in Visul Corticl nsemle hengjin Xu, Wnchen Jing, Mu-ming Poo, nd Yng Dn

2 c 1 nd point ( ) A P L V1 V2M V1M V1B V2L λ trting point ( ) d Firing rte (spikes per second) Time (s) upplementry Figure 1 RFs nd conditioning-evoked sequentil spiking of neurons in nesthetized rt V1. () chemtic of experimentl setup. Visul stimuli were presented to left eye, multielectrode rry (red dots) ws inserted into right V1. Digrm of rt cortex ws dopted from Pxinos nd Wtson 48, shown on 1 1 mm grid. () Multiunit RFs recorded simultneously in urethne-nesthetized rt. Red ellipse, contour of Gussin fit t one stndrd devition. (c) uperposition of RFs nd visul stimuli. Colored ellipses, Gussin fits of RFs. White dshed circles, trting point () nd nd point () of conditioning moving spot. (d) PTHs of the units during conditioning stimultion y moving spot, ordered y distnce etween RF center nd. Red curves, PTHs smoothed with Byesin dptive regression splines. -1-

3 ynch Desynch Horizontl Verticl 6 s 5 degree Percentge Horizontl ynch Desynch 2 ynch Desynch 1 Degree Verticl Degree Percentge upplementry Figure 2 ye movement in wke hed-fixed rts. () xmple eye movement trces from trined rt. Red nd lue lines, periods of synchronized nd desynchronized rin sttes (sme s in Fig. 6). () Distriution of eye positions recorded in 5 experiments during test periods from 2 trined rts, in synchronized (red) nd desynchronized (lue) rin sttes. uperimposed is 2-D Gussin function (luminnce profile nd gry dshed lines) whose size is mtched to the verge RF size of rt V1 neurons mesured in our experiments. -2-

4 Cumultive % 1 5 Anesthetized Cumultive % 1 5 Awke upplementry Figure 3 Alterntive nlysis of spike sequence evoked y test cue t in nesthetized nd wke rts. hown re the cumultive histogrms of permn s etween test stimulus -evoked spiking sequence nd conditioning stimulus-evoked spiking sequence, efore (dotted line) nd fter (solid line) 1 trils of conditioning t 18 s 1 for nesthetized (, n = 19 experiments) nd wke (, n = 18) rts. Conditioning cused significnt rightwrd shift of the distriution (, P = ;, P = , Kolmogorov-mirnov test). -3-

5 Cumultive % 1 5 Cond Test Anesthetized ( ) Cumultive % 1 5 Anesthetized (flshed spot, ) Cond Test c Cumultive % 1 5 Anesthetized (flshed spot, ) Cond Test upplementry Figure 4 Cumultive histogrms of permn s for control experiments in nesthetized rts. () Cumultive histogrms of s for test stimuli t following 1 trils of conditioning. ws computed etween test stimulus-evoked spiking sequence nd RF position long the xis, efore (dotted line) nd fter (solid line) 1 trils of conditioning t 18 s 1 for nesthetized rts (n = 19 experiments). There ws no significnt difference efore nd fter conditioning (P =.77, Kolmogorov-mirnov test). () Cumultive histogrms of s for test stimuli t following 1 trils of flshed-spot conditioning t (n = 17 experiments). There ws no significnt difference efore nd fter conditioning (P =.47). (c) Cumultive histogrms of s for test stimuli t following 1 trils of flshed-spot conditioning t. There ws no significnt difference efore nd fter conditioning (P =.44). Digrm in upper left region of ech plot illustrtes conditioning nd test stimuli used in tht experiment. -4-

6 1 trting point ( ) nd point ( ) trting point ( ) c nd point ( ) 1 1 Cumultive % 5 Cumultive % upplementry Figure 5 Control experiment with the conditioning spot moving long motion pth prllel to, ut not overlpping with, the long xis of the recorded RF distriution. () uperposition of RFs nd visul stimuli. Colored ellipses, Gussin fits of RFs. White filled circle nd rrow indicte conditioning motion pth. White dshed circles, trting point () nd nd point () of test spot. () Cumultive histogrms of permn s etween -evoked spike sequence nd RF position, efore (dotted line) nd the first 2 min fter (solid line) conditioning (n = 14 experiments). There ws no significnt difference (P =.8; Kolmogorov-mirnov test). (c) Cumultive histogrms of permn s for -evoked spike sequence. The difference ws not significnt (P =.93). -5-

7 1 A L P nd point ( ) V2M V1M V1 V1B V2L λ trting point ( ) c RF distnce (degrees) Conditioning.5.5 efore Time (s) 1.5 d Cumultive % 1 5 trting point ( ) Cond Test e Cumultive % 1 5 Cond Test nd point ( ) upplementry Figure 6 Conditioning-induced increse of sequentil spiking in nesthetized rts, with electrode rry implnted t different ngle. () chemtic of experimentl setup. Multielectrode rry (red dots) ws inserted into right V1. Digrm of rt cortex ws dopted from Pxinos nd Wtson 48, shown on 1 1 mm grid. () uperposition of RFs nd visul stimuli. Colored ellipses, Gussin fits of RFs. White dshed circles, trting point () nd nd point () of conditioning spot. (c) Top three rows, pirwise cross-correltion verged from ll experiments with this electrode rry orienttion (n = 18 experiments, including 8 experiments with 2 trils nd 1 experiments with 5 trils of conditioning). Bottom row, difference etween cross-correltion functions efore nd fter conditioning. (d) Cumultive histogrms of permn s etween -evoked spike sequence nd RF position, efore (dotted line) nd the first 2 min fter (solid line) conditioning for ll 18 experiments. The difference ws significnt (P = ; Kolmogorov-mirnov test). (e) Cumultive histogrms of permn s for -evoked spike sequence. The difference ws not significnt (P =.53). -6-

8 % of mtches % of mtches threshold c Time (min) 1 Cumultive % upplementry Figure 7 Persistence of conditioning-induced increse in sequentil spiking in nesthetized rts fter 1 trils of conditioning t 18 s 1. () Difference in the percentge of sequence mtches efore nd fter conditioning vs. threshold for test stimuli t (lck) nd (gry). The difference ws significnt t ll thresholds elow.9 for ( *, P <.5; **, P <.1; ***, P <.1; Wilcoxon signed rnk test), ut not t ny threshold for. rror r, s.e.m. () Time course for decy of conditioning-induced increse in the percentge of mtches (t threshold of.6). For, the increse ws significnt t ll time points (2 min, P = ; 4 min, P =.44; 6 min, P =.27; Wilcoxon signed rnk test) fter conditioning. For, the effect ws not significnt t ny time. rror r, s.e.m. (c) Cumultive histogrms of permn s t different periods fter conditioning (solid lines). Dotted line, histogrm efore conditioning. The difference is significnt t 2 min (P = ; Kolmogorov-mirnov test) nd 4 min (P =.13), ut not t 6 min (P =.79). -7-

9 Anesthetized Anesthetized Awke Cond speed (36 s 1 ) Cond speed (18 s 1 ) Cond speed (18 s 1 ) Count 4 Count 4 Count Recll speed (1 s 1 ) Recll speed (1 s 1 ) Recll speed (1 s 1 ) Anesthetized Awke 1 µv 1 s 2 µv 1 s upplementry Figure 8 peed of cue-triggered recll of spike sequence in nesthetized nd wke rts. () Left, histogrms of speed for -triggered recll of spike sequences in ll nesthetized experiments (n = 18) with conditioning speed t 36 s 1. Only mtched trils with permn >.9 were included in this nlysis. Arrow, men. Middle, histogrms of recll speed in ll nesthetized experiments with 1 trils of conditioning t speed of 18 s 1 (n = 19). Right, histogrms of recll speed in ll wke experiments (n = 18) with conditioning speed t 18 s 1. () xmple LFP trces recorded from urethne-nesthetized nd n wke rt. -8-

10 Men firing rte (spikes per second) 2 1 locl ppliction of APV (75 µm) Time (s) upplementry Figure 9 Decrese in corticl firing rte fter locl ppliction of APV. ch red circle represents men spontneous firing rte over period of 1 s, verged cross ll units recorded in the 18 APV experiments. The men firing rte decresed to 57 ± 7% (s.e.m.) of the seline nd ecme stle within 1 min fter the APV ppliction. xperiments on sequence recll were performed ~ 3 min fter APV ppliction, when the firing rte ws stilized. hdow re, period of APV ppliction. -9-

11 upplementry Movie. 1 Awke hed-fixed rt during visul stimultion. Red circle indictes pupil size nd position. Arrows point to reflections of the test stimuli ( nd, presented with n LCD monitor in front of the left eye) off the eye ll. The sttionry right spot on the left of the eye ws the reflection of the infr-red light source tht provides illumintion for the cmer. Horizontl nd verticl eye positions re shown in the trces elow. Also shown re simultneously recorded LFP, together with red/lue lines indicting synchronized/desynchronized rin sttes, nd the short rs elow indicting periods of visul stimultion. Note tht the desynchronized rin stte is ssocited with some fcil nd whisker movement. -1-