Extended Data Fig. 9: Selective PKA inhibition in SPNs slows learning.

a, Schematic describing a strategy to investigate the effect of D1R-SPN or D2R-SPN PKA inhibition on behaviour. AAV1-FLEX-PKIalpha-IRES-nls-mRuby2 was injected into the NAc of Drd1a-cre or Adora2a-cre mice to selectively inhibit PKA in D1R-SPN or D2R-SPN, respectively. For control groups, AAV1-Cag-FLEX-eGFP was injected instead. Ten to fourteen days after surgery, mice were started on a behaviour schedule that includes 1 day of habituation (day 0) and 11 days of training (days 1 to 11). b, Effect of D1R-SPN PKA inhibition on the fraction of time spent in trigger zone (time spent in trigger zone/total session time). Daily average time (left, two-way repeated-measures ANOVA day × group interaction, F(10, 140) = 5.565, P < 0.0001) and multiday average (right, unpaired t-test, P = 0.012, 0.017, 0.323). c, Effect of D1R-SPN PKA inhibition on the speed after LED (average speed during 0–1.2 s after LED onset; 1.2 s is the minimum latency to enter the receptacle zone after LED cue). Daily average speed after LED onset (left, two-way repeated-measures ANOVA day × group interaction, F(10, 140) = 2.923, P = 0.002) and multiday average (right, unpaired t-test, P = 0.046, >0.999, >0.999). d, Effect of D1R-SPN PKA inhibition on the entering failure rate (number of entering failure trials/total number of trials). Daily average entering failure rate (left, two-way repeated-measures ANOVA day × group interaction, F(10, 140) = 2.591, P = 0.007) and multiday average (right, unpaired t-test, P = 0.043, 0.367, 0.073). e, Effect of D2R-SPN PKA inhibition on the time spent in trigger zone. Daily average time (left, two-way repeated-measures ANOVA day × group interaction, F(10, 140) = 1.322, P = 0.224) and multiday average (right, unpaired t-test, P > 0.999, >0.999, >0.999). f, Effect of D2R-SPN PKA inhibition on the speed after LED. Daily average speed after LED onset (left, two-way RM ANOVA day × group interaction, F(10, 140) = 3.124, P = 0.001) and multiday average (right, unpaired t-test, P = 0.938, 0.011, 0.145). g, Effect of D2R-SPN PKA inhibition on the entering failure rate. Daily average entering failure rate (left, two-way repeated-measures ANOVA day × group interaction, F(10, 140) = 1.951, P = 0.043) and multiday average (right, unpaired t-test, P > 0.999, = 0.011, >0.999). h, Occupancy failure rate (number of premature receptacle zone exit trials/number of receptacle zone entering success trials) of D1R-SPN PKA inhibition experiments (one-way repeated-measures ANOVA on GFP group, F(3.213, 22.49) = 1.918, P = 0.153; one-way repeated-measures ANOVA on PKI group, F(2.424, 16.97) = 0.352, P = 0.747). i, Left, average speed (total distance travelled/total session time, two-way repeated-measures ANOVA group effect, F(1, 14) = 6.506, P = 0.023). Right, average speed during baseline period (−20 s before trigger zone entry to trigger zone entry, two-way repeated-measures ANOVA group effect, F(1,14) = 4.304, P = 0.057). j, DAPI cell counting for D1R SPN PKA inhibition (unpaired t-test, P = 0.517). k, Occupancy failure rate of D2R-SPN PKA inhibition experiments (one-way repeated-measures ANOVA on GFP group, F(2.293, 16.05) = 2.062, P = 0.155; one-way repeated-measures ANOVA on PKI group, F(1.598, 11.19) = 1.482, P = 0.264). l, Left, average speed (two-way repeated-measures ANOVA group effect, F(1, 14) = 0.011, P = 0.916). Right, average speed during baseline period (two-way repeated-measures ANOVA group effect, F(1, 14) = 0.211, P = 0.653). m, DAPI cell counting for D2R SPN PKA inhibition (unpaired t-test, P = 0.598). *P < 0.05 for Bonferroni-corrected unpaired t-tests. All t-tests are two-sided. Individual dots on the bar graph = a mouse. All graphs are plotted as mean ± s.e.m. across mice (n = 8 for each group).