Transient Stimulation of Distinct Subpopulations of Striatal Neurons Mimics Changes in the Value of Competing Actions

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1 Supplementl mterils for: Trnsient Stimultion of Distinct Supopultions of Stritl Neurons Mimics Chnges in the Vlue of Competing Actions Lung-Ho Ti, A. Moses Lee,2, Nor Benvidez 3, Antonello Bonci 4,,6, Lind Wilrecht,4 Ernest Gllo Clinic nd Reserch Center, Emeryville, CA 2 Medicl Scientist Trining Progrm, Neuroscience Grdute Progrm, University of Cliforni, Sn Frncisco 3 Deprtment of Cognitive Science, University of Cliforni, Berkeley 4 Deprtment of Neurology, University of Cliforni, Sn Frncisco Intrmurl Progrm, Ntionl Institute of Drug Ause, Bltimore, MD 6 Solomon H. Snyder Deprtment of Neuroscience, Johns Hopkins School of Medicine These uthors mde equl contriutions. Nture Neuroscience: doi:.38/nn.388

2 Supplementry Figure. Chosen port: Left Right Frction left choice Withdrwl time (ms) Frction rewrd trils t left port Reltive ction vlue for left choice c Frction of totl trils. 2 2 Go-Cue to nose-out (ms) d Movement time (ms) Chosen port: Left Right Reltive ction vlue for left choice e. f... Reltive ction vlue. Predicted. Supplementry Figure. Chrcteriztion of responses within the tsk () The reltionship of frction of left choices nd frction of rewrd trils t left port for ll 6 possile rewrd histories in the previous two trils verged cross ll sujects. The frequency of trils of given rewrd history is indicted y the reltive size of the circle. () Averge medin withdrwl time from gosignl to nose-withdrwl t center port (n=28). Withdrwl time is shorter when the ction vlue for the chosen port is higher. (c) Cumultive distriution of rection time from Go-Cue to nose-withdrwl from center port for ll sujects (men ±s.d., n=28). The dotted line indictes the onset of opticl ultion t ms ltency. (d) Averge movement time from nose-withdrwl t center port to rewrd port (n=28). Movement time is shorter when the ction vlue for the chosen port is higher. All error rs represent s.e.m. (e) The frction of choices for the left port from 3% of dt given the reltive ction vlue eted from the other 7% of dt. Dt from ech suject were grouped in ins nd represented in distinct color. (f) The frction of choices for the left port from 3% of dt given the frction of left choices predicted y the regression model using the other 7% of dt. Nture Neuroscience: doi:.38/nn.388

3 Supplementry Figure 2 D-ChR2-eYFP D-eYFP +.3±.2 mm +.7±.2 mm +.±.2 mm eyfp control c D2-ChR2-eYFP ChR2-eYFP D2-eYFP d +.3±.2 mm +.7±.2 mm +.±.2 mm Supplementry Figure 2. Antomy of ultion sites () Coronl series demonstrting the extent of infection (grey) nd plcements of fier optic tips (dots) for D-Cre injected with AAV2/-EF-DIO-ChR2-eYFP (left series) or AAV2/-EF-DIO-eYFP (right) series. Light grey represents the extent of the lrgest injection while drk grey represents the smllest extent. () Coronl (top pnel) nd sgittl histologicl sections (ottom pnel) from two representtive D-Cre nimls expressing ChR2-eYFP. (c) Coronl series demonstrting the extent of infection (grey) nd plcements of fier optic tips (dots) for D2-Cre injected with AAV2/-EF-DIO-ChR2-eYFP (left series) or AAV2/-EF-DIO-eYFP (right) series. (d) Coronl (top pnel) nd sgittl histologicl sections (ottom pnel) from two representtive D2-Cre nimls expressing ChR2eYFP. Nture Neuroscience: doi:.38/nn.388

4 Supplementry Figure 3 D-Cre c D2-Cre D-Cre d Supplementry Figure 3. Expression of ChR2 in Stritl MSNs nd ChAT+ Interneuons () Single plne confocl imges of medium spiny neurons nd cholinergic interneurons in the dorsomedil stritum from D-Cre mouse injected with AAV-EFα-DIO-ChR2-eYFP (ChR2-eYFP, green in merged imge). Histologicl slices were leled using immunohistochemistry for the intrcellulr C-terminus of Kv2. chnnel, mrker of MSNs 2 (red in merged imge), nd choline cetyltrnsferse (ChAT, lue in merged imge) respectively. ChR2-eYFP expression in D-Cre nimls ws found to coloclize with Kv2. expression s indicted y the red rrows in (), ut never with ChAT positive neurons (/68 neurons). () Single plne confocl imges of medium spiny neurons nd cholinergic interneurons in the dorsomedil stritum from D2-Cre mouse injected with AAV- EFα-DIO-ChR2-eYFP. ChR2-eYFP expression in D2-Cre nimls ws found to coloclize with Kv2. expression (43/4 neurons) s indicted y the red rrows in mny cells ( nd c). In D2-Cre nimls, ChR2-eYFP expression ws found to coloclize with suset of ChAT immunostined neurons (29/7) (lue dot in ()), ut not ll ChAT immunostined neurons (lue dot in (c)) despite the presence of ChR2-eYFP expression in nery cells s seen y 3D-reconstruction (d). Scle r in (c): μm. Scle r in (d): 2 μm Nture Neuroscience: doi:.38/nn.388

5 Supplementry Figure 4 Light ultion Tril # Normlized spike mplitude D-ChR n= Time (ms) Tril # Tril # Tril # Time (sec) Time (sec) Normlized spike mplitude Normlized Spike Amplitude Normlized spike mplitude D-ChR n= Time (ms) D2-ChR2 - n= Time (ms) D2-ChR n= Time (ms) Supplementry Figure 4. Opticl ultion induces spiking in ChR2-trnsduced stritum () Spike rster of two representtive single units for light-evoked ctivity in the stritum of D-ChR2 mice. Stimultion ws delivered with ms pulses t 2 Hz for ms. The wveforms of the units re shown in the right pnels. () Spike rster of two representtive single units for light-evoked ctivity in the stritum of D2-ChR2 mice. Right: recorded spike wveforms. Nture Neuroscience: doi:.38/nn.388

6 Supplementry Figure Hz (3 pulses) Hz ( pulses) 2 Hz ( pulses) Hz (3 pulses) Hz ( pulses) 2 Hz ( pulses) D D D D D Reltive ction vlue.. - D Reltive ction vlue Supplementry Figure. Gllery of individul D-ChR2 nimls dt Frction of choices for the left port on trils with different reltive ction vlue etes in D-cre nimls in the presence (red) or sence (lue) of opticl ultion (protocol in Fig. 3). All sujects were trnsduced with AAV-EFα-DIO-ChR2-eYFP. Opticl ultion in the right nd left hemisphere re shown seprtely. Some mice only demonstrted ChR2 expression unilterlly nd ultion sessions were only used from the trnsduced side. Logistic regression ws used to fit the dt from trils with (red curve) nd without ultion (lue curve). All error rs represent s.e.m. Nture Neuroscience: doi:.38/nn.388

7 Supplementry Figure 6 Hz (3 pulses) Hz ( pulses) 2 Hz ( pulses) Hz (3 pulses) Hz ( pulses) 2 Hz ( pulses).. - D D D D D D D D Reltive ction vlue Reltive ction vlue Supplementry Figure 6. Gllery of individul D2-ChR2 nimls dt Frction of choices for the left port on trils with different reltive ction vlue etes in D2-cre nimls in the presence (red) or sence (lue) of opticl ultion (protocol in Fig. 3). Dt from ll sujects trnsduced with AAV-EFα-DIO-ChR2-eYFP with opticl ultion in the right nd left hemisphere. Logistic regression ws used to fit the dt from trils with (red curve) nd without ultion (lue curve). All error rs represent s.e.m. Nture Neuroscience: doi:.38/nn.388

8 Supplementry Figure 7 Prev. choice: contrlterl Current Choice: contrlterl Withdrwl time (ms) D DMS (ChR2-ChR2-eYFP summry dt, n= ultion sites, 6 nimls) Prev. choice: ipsilterl Current Choice: ipsilterl Prev. choice: contrlterl Current Choice: contrlterl Withdrwl time (ms) Withdrwl time (ms) D2 DMS (ChR2-ChR2-eYFP summry dt, n=3 ultion sites, 8 nimls) Reltive ction vlue for ipsilterl choice Prev. choice: ipsilterl Current Choice: ipsilterl Withdrwl time (ms) Reltive ction vlue for ipsilterl choice Supplementry Figure 7. Opticl ultion ltered withdrwl time on trils when nimls did not switch sides On trils when nimls did not switch sides reltive to the previous tril (sty trils) we plot time to withdrw from center port fter the go signl (withdrwl time) ginst port choice on trils with different reltive ction vlue etes. Two different lines reflect the presence (red) or sence (lue) of opticl ultion (protocol in Fig. 3). () shows tht ultion speeds contrlterl choice nd slows ipsilterl choice withdrwl time in D-cre mice. () Shows ultion speeds ipsilterl choice nd slows contrlterl choice withdrwl time in D2-cre mice. Note we plot only dt points where more thn % of sujects hve or more trils. All dt is from sujects trnsduced with AAV-EFα- DIO-ChR2-eYFP. All error rs represent s.e.m. : p<.; : p<., Wilcoxon signed-rnk test. Nture Neuroscience: doi:.38/nn.388

9 Supplementry Figure 8 D-9, left DMS. D-22, left DMS. D-22, right DMS.... p< 3 p< 29 Reltive ction vlue Reltive ction vlue p< 3 Reltive ction vlue D2-3, left DMS. D2-4, left DMS. D2-, left DMS. D2-6, left DMS..... p< 47 p< 3 p< 8 p< 2 Reltive ction vlue Reltive ction vlue Reltive ction vlue Reltive ction vlue c Withdrwl time (ms) 2 D-ChR2 DMS (n=3 sites, 2 nimls) Reltive ction vlue for ipsilterl choice d 2 D2-ChR2 DMS (n=4 sites, 4 nimls) Reltive ction vlue for ipsilterl choice Supplementl Figure 8. Individul nimls dt for pre go-signl ultion experiment (-) Frction of choices for the left port on trils with different reltive ction vlue etes in () D- cre or () D2-cre nimls in the presence (red) or sence (lue) of opticl ultion efore gosound (protocol shown in Fig 7). Logistic regression ws used to fit the dt from trils with (red curve) nd without ultion (lue curve). P vlues reported for t-tests: H :ß = (distnce etween red nd lue lines). (c-d) The medin time tken to withdrw from the center port verged cross individul D-Cre sujects (c) nd D2-Cre sujects (d) in trils without ultion (lue) or with ultion (red) nd cross different reltive ction vlues for choosing the port ipsilterl versus contrlterl to the site of ultion. Positive reltive ction vlues correspond to trils in which the vlue of the port ipsilterl to the site of ultion is greter thn the contrlterl port. All dt is from sujects trnsduced with AAV-EFα-DIO-ChR2-eYFP. : p<., Wilcoxon signed-rnk test. All error rs represent s.e.m. Nture Neuroscience: doi:.38/nn.388

10 Supplementry Figure 9 Center-port-in Go signl In proilistic switching tsk Frme: Frme: Frme: (/3 sec) (/6 sec) Rewrd port in Frme: Frme: (/2 sec) Red: initil ody orienttion Frme: 9 (3/ sec) Frme: 3 (3/3 sec) Frme: (/2 sec) Yellow: turning degree clockwise Blue: turning 7 degree clockwise c Frme: (ultion onset) Outside tsk context trnsient ultion d Frme: (/2 sec) Normlized ody turning (degree) - D-ChR2 (n=8 sites,4 mice) D2-ChR2 (n=2 sites,7 mice). sec sec. sec sec fter trnsient ultion (2 Hz ms) Ipsilterl is Contrlterl is Outside tsk context prolonged ultion e D-ChR2 (n=9 sites, mice) f D2-ChR2 (n= sites, 7 mice) Bseline (6 s) Stimultion (6 s) Recovery (6 s) Bseline (6 s) Stimultion (6 s) Recovery (6 s) # of contrlterl rottion per minute - ^ ^^^ ^^^ Hz, 6 sec Hz, 6 sec 2 Hz, 6 sec # of contrlterl rottion per minute - - ^^ Bseline vs. ultion : p<. : p<. : p<. Wilcoxon rnk sum test ^^^ ^^ Stimultion vs. recovery ^: p<. ^^: p<. ^^^ p<. Wilcoxon rnk sum test Nture Neuroscience: doi:.38/nn.388

11 Supplementry Figure 9. Effect of ultion on ody orienttion nd rottion outside of the tsk context () A series of video frmes showing the time course of typicl tril. () Two top-view video frmes showing the typicl ody orienttion t center-port-in nd. sec lter. (c) Two top-view video frmes showing the typicl ody orienttion t ultion onset nd. sec lter. Red dot: eted center of ody. (d) Normlized ody turning in. sec or sec fter rief ms 2 Hz stritl ultion in D-ChR2 (p=.46 nd p=.74, n=8) nd D2-ChR2 nimls (p=.79 nd p=.9, n=2) outside the ehvior tsk. P vlues reported for Wilcoxon sign rnk test. (e) The numer of contrlterl nd ipsilterl rottions in the seline, prolonged ultion nd recovery periods from D-ChR2 nimls. The durtion of ech period ws 6 seconds. (f) The numer of contrlterl nd ipsilterl rottions in the seline, prolonged ultion nd recovery periods from D2-ChR2 nimls. Reported n refers to numer of ultion sites. All error rs represent s.e.m. Nture Neuroscience: doi:.38/nn.388

12 Supplementry Figure log( P Models: ) = Z + β X Akike Informtion Criterion (AIC) : L AIC = ± 3.2 PL log( P L ) = β Signed-rnk Test P-Vlue 2: AIC 2 = 69.6 ± 3.23 H : P AIC = AIC 2.26 L Z + β 3 PL Re wrd 3: log( ) = β j ( YL ( i j) YR ( i j)) + AIC 3 = 7.84 ± 3.67 H : PL j = AIC = AIC 3.3 n j = β No Re wrd j ( N L ( i j) N R 4: P AIC 4 = ± 3.7 L = ( Z + β X ( i )) + e : AIC = 7.69 ± 4.4 H : ( Z + β X ( i )) AIC 4 = AIC.9 + e 6: L AIC 6 = ± 2.3 H : ( Z + β X ( i )) AIC 4 = AIC : L AIC 7 = 7.7 ± 2.9 H : ( Z + β X ( i )) / AIC 4 = AIC Z is the eted reltive ction vlue for no-ultion trils with the sme pst rewrd nd choice history. :AIC clculted with correction 2k(k+)/(n-k-) for finite smple size. AIC vlue reported: medin ± stndrd error of medin clculted using ootstrp smpling ( i X j)) + Z + β P L i = ( ) / P = + + e P = + + n j = e y n Re wrd No Re wrd j ( YL ( i j) YR ( i j)) + β j ( N L ( i j) N R ( i j)) β j = Z = β + y X D D2 c D D2 Chnge in reltive ction vlue D-ChR2 (n=) D-eYFP (n=7) Hz Hz 2Hz Hz Hz 2Hz D2-ChR2 (n=3) D2-eYFP (n=7) Slope et coefficient.4 Hz Hz 2 Hz Hz Hz 2 Hz.2.8 p=.4 p=.383 p=.28 p= p= p=.3 D-ChR2 (n=) D2-ChR2 (n=3) D-eYFP (n=7) D2-eYFP (n=7) Nture Neuroscience: doi:.38/nn.388

13 Supplementry Figure. Comprison of lterntive models in descriing the nimls ehvior () In order to rule out whether other lterntive models could e used to descrie the nimls ehvior, we generted fmily of more complex models. In these models, we compre our simple generlized liner model (model ) with: Model 2: A model tht llows for ultion to chnge the generl sensitivity to the ction vlue. Model 3: A model tht llows for ultion to chnge the sensitivity to the pst rewrd history in the lst three trils. Model 4: A nonliner fit model similr to our generlized liner model tht llows for shift. Model : A nonliner fit model tht llows for ultion to chnge the generl sensitivity to the ction vlue. Model 6: A nonliner fit model tht llows for chnge in the upper nd lower ound symptotes. Model 7: A nonliner fit model tht llows for chnge in the symptotes s well s generl chnge in sensitivity to the ction vlue. This model ws sed upon method used y Erlich, Bilek, nd Brody. Neuron. 2. We then compred ll of these models to our generlized liner model to determine whether ny of these models could more ccurtely descrie our dt given the dditionl free prmeters tht they require using the Akike Informtion Criterion (AIC). Bsed upon this model selection criterion, we found tht these clsses of more complex models were not significntly different or performed worse thn our simple generlized liner model in terms of their goodness-of-fit. Given its simplicity, we elieve tht descriing the effect of ultion s shift in the ction vlue is vlid nd ccurte mesure for descriing our dt. P vlues reported for Wilcoxon signed-rnk test. Exmple nlysis from Model 2. We nlyzed the effect of opticl ultion using model 2 which llows the ultion to cuse shift in reltive ction vlue (ß ) s well s chnges in nimls sensitivity to rewrd (slope, ß ). () Eted chnge in the reltive ction vlue (shift, ß ) for choosing the port ipsilterl versus contrlterl to the side of ultion verged cross individuls within group. Positive chnges in reltive ction vlues correspond with n ipsilterl is while negtive chnges correspond to contrlterl is. (c) Eted chnge in rewrd sensitivity (slope, ß ) verged cross individuls within group. No significnt chnge in rewrd sensitivity ws oserved. Reported n refers to numer of ultion sites. : p<., P vlues reported for Wilcoxon rnk-sum test. All error rs represent s.e.m. Nture Neuroscience: doi:.38/nn.388

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