Anterograde tracing in D1-Cre and A2a-Cre rats

A. D1-Cre or A2a-Cre rats were unilaterally infused in the dorsal striatum (DS) or nucleus accumbens shell (NAc-S) with DIO-eYFP. B. Sagittal micrographs obtained in D1-Cre (top) and A2A-Cre (bottom) rats following viral infusion in the DS. C. DIO-eYFP infusion in the DS of D1-Cre rats. Micrographs show eYFP expression in the DS, D32 staining and co-labelling (eYFP + D32) in the DS. They also show that DS D1-SPNs project to the substantia nigra pars reticulata (SNr) and the globus pallidus externus (GPe). Viral expression was restricted to putative SPNs (D32 | eYFP), with ∼40% of SPNs expressing eYFP (eYFP | D32). D. DIO-eYFP infusion in the DS of A2a-Cre rats. Micrographs show eYFP expression in the DS, D32 staining and co-labelling (eYFP + D32) in the DS. They also show that DS D2-SPNs project to the SNr but not the GPe. Viral expression was restricted to putative SPNs (D32 | eYFP), with ∼40% of SPNs expressing eYFP (eYFP | D32). E. DIO-eYFP infusion in the NAc-S of D1-Cre rats. Micrographs show eYFP expression in the NAc-S, D32 staining and co-labelling (eYFP + D32) in the DS. They also show that NAc-S D1-SPNs project to the ventral pallidum (VP) and the lateral hypothalamus (LH). Viral expression was restricted to putative SPNs (D32 | eYFP), with ∼41% of SPNs expressing eYFP (eYFP | D32). F. DIO-eYFP infusion in the NAc-S of A2a-Cre rats. Micrographs show eYFP expression in the NAc-S, D32 staining and co-labelling (eYFP + D32) in the DS. They also show that NAc-S D2-SPNs project to the VP but not the LH. Viral expression was restricted to putative SPNs (D32 | eYFP), with ∼40% of SPNs expressing eYFP (eYFP | D32).

Ex-vivo cell recordings in D1-Cre and A2a-Cre rats

A. D1-Cre were bilaterally infused in the NAc-S with DIO-eYFP (black) or DIO-eNpHR3.0 (blue). The representative raw traces of cell attached recordings are those of transfected neurons that were depolarized to elicit action potentials by injecting a brief positive current step (+150pA, 200ms duration, 0.5Hz). 625nm LED illumination (orange bar, continuous wave, 2mW) had no effect in eYFP transfected neurons (black) but it inhibited action potential in eNpHR3.0 transfected neurons (blue). The grouped data for recordings include 5 animals (overlapping data points). B. A2a-Cre were bilaterally infused in the NAc-S with DIO-eYFP (black) or DIO-eNpHR3.0 (red). The representative raw traces of cell attached recordings are those of transfected neurons that were depolarized to elicit action potentials by injecting a brief positive current step (+150pA, 200ms duration, 0.5Hz). 625nm LED illumination (orange bar, continuous wave, 2mW) had no effect in eYFP transfected neurons (black) but it inhibited action potential in eNpHR3.0 transfected neurons (red). The grouped data for recordings include 7 animals (overlapping data points).

NAc-S D1-SPNs mediate outcome-specific PIT

A. D1-Cre rats were bilaterally infused in the NAc-S with DIO-eYFP (black) or DIO-eNpHR3.0 (blue). Fiber-optic cannulas were implanted above the NAc-S to provide 625nm LED illumination B. Schematic representation of the behavioral design; S1 and S2: noise and clicker stimuli (counterbalanced); O1 and O2: grain pellets and sucrose solution (counterbalanced); A1 and A2: left and right lever press (counterbalanced). At test, S1 and S2 were presented 4 times each, in a pseudorandom order. Half of the trials for each stimulus was conducted under 625nm LED illumination (ON; continuous wave; ∼10mW) whereas the LED remained inactivated during the other half of the trials (OFF). ON/OFF trials were counterbalanced. C. Outcome-specific PIT test: net lever presses when the stimuli predicted the same outcome as the action (Same) or when the stimuli predicted the different outcome (Different). Lever presses are shown for each group in trials conducted under 625nm LED illumination (ON) and in trials without illumination (OFF). Data are shown as mean ± SEM. Panel C includes individual data points for female (filled circle) and male (open circle) rats. Asterisks denote significant effect (* p < 0.05; ** p < 0.01; *** p < 0.001; n.s., nonsignificant).

NAc-S D2-SPNs mediate for outcome-specific PIT

A. A2a-Cre rats were bilaterally infused in the NAc-S with DIO-eYFP (black) or DIO-eNpHR3.0 (red). Fiber-optic cannulas were implanted above the NAc-S to provide 625nm LED illumination. B. Outcome-specific PIT test: net lever presses when the stimuli predicted the same outcome as the action (Same) or when the stimuli predicted the different outcome (Different). Lever presses are shown for each group in trials conducted under 625nm LED illumination (ON) and in trials without illumination (OFF). Data are shown as mean ± SEM. Panel B includes individual data points for female (filled circle) and male (open circle) rats. Asterisks denote significant effect (* p < 0.05; ** p < 0.01; n.s., nonsignificant).

NAc-S D1-SPNs projections to the VP mediate outcome-specific PIT

A. D1-Cre rats were bilaterally infused in the NAc-S with DIO-eYFP (black) or DIO-eNpHR3.0 (blue). Fiber-optic cannulas were implanted above the VP to provide 625nm LED illumination B. Outcome-specific PIT test: net lever presses when the stimuli predicted the same outcome as the action (Same) or when the stimuli predicted the different outcome (Different). Lever presses are shown for each group in trials conducted under 625nm LED illumination (ON) and in trials without illumination (OFF). Data are shown as mean ± SEM. Panel B includes individual data points for female (filled circle) and male (open circle) rats. Asterisks denote significant effect (** p < 0.01; *** p < 0.01; n.s., nonsignificant).

NAc-S D2-SPNs projections to the VP do not mediate outcome-specific PIT

A. A2a-Cre rats were bilaterally infused in the NAc-S with DIO-eYFP (black) or DIO-eNpHR3.0 (red). Fiber-optic cannulas were implanted above the VP to provide 625nm LED illumination. B. Outcome-specific PIT test: net lever presses when the stimuli predicted the same outcome as the action (Same) or when the stimuli predicted the different outcome (Different). Lever presses are shown for each group in trials conducted under 625nm LED illumination (ON) and in trials without illumination (OFF). Data are shown as mean ± SEM. Panel B includes individual data points for female (filled circle) and male (open circle) rats.