The influence of nucleus accumbens shell D1 and D2 neurons on outcome-specific Pavlovian instrumental transfer
- Decision Neuroscience Laboratory, School of Psychology, Australia
Figures
Figure 1
Anterograde tracing in D1-Cre and A2a-Cre rats.
(A) D1-Cre or A2a-Cre rats were unilaterally infused in the dorsal striatum (DS; D1-Cre: n = 4 males; A2A-Cre: n = 3 males) or nucleus accumbens shell (NAc-S; D1-Cre: n = 1 female and 1 male; A2A-Cre: n = 1 female and 1 male) 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-labeling (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-labeling (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-labeling (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-labeling (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).
Figure 2
Ex vivo cell recordings in D1-Cre and A2a-Cre rats.
(A) D1-Cre was bilaterally infused in the NAc-S with DIO-eYFP (black; 2 females) or DIO-eNpHR3.0 (blue; eNpHR3.0; 2 females). 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 (+150 pA, 200 ms duration, 0.5 Hz). 625 nm LED illumination (orange bar, continuous wave, 2 mW) had no effect in eYFP transfected neurons (black; 5 cells) but it inhibited action potential in eNpHR3.0 transfected neurons (blue; 7 cells). The grouped data for recordings include overlapping data points. (B) A2a-Cre was bilaterally infused in the NAc-S with DIO-eYFP (black; 2 females) or DIO-eNpHR3.0 (red; 2 females). 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 (+150 pA, 200ms duration, 0.5 Hz). 625 nm LED illumination (orange bar, continuous wave, 2 mW) had no effect in eYFP transfected neurons (black; 8 cells) but it inhibited action potential in eNpHR3.0 transfected neurons (red; 6 cells). The grouped data for recordings include overlapping data points.
Figure 3 with 1 supplement
NAc-S D1-SPNs mediate outcome-specific Pavlovian instrumental transfer (PIT).
(A) D1-Cre rats were bilaterally infused in the NAc-S with DIO-eYFP (black; 5 females and 5 males) or DIO-eNpHR3.0 (blue; 5 females and 5 males). Fiber-optic cannulas were implanted above the NAc-S to provide 625 nm LED illumination (continuous). (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 the test, S1 and S2 were presented four times each, in a pseudorandom order. Half of the trials for each stimulus was conducted under 625 nm LED illumination (ON; continuous wave; ~10 mW) 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 625 nm 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).
Figure 3—figure supplement 1
Histological and behavioral data related to Figure 3.
D1-Cre rats were bilaterally infused in the NAc-S with DIO-eYFP (black; 5 females and 5 males) or DIO-eNpHR3.0 (blue; 5 females and 5 males). (A) Minimal (light gray for eYFP group and light blue for eNpHR3.0 group) and maximal (darker gray for eYFP group and darker blue for eNpHR3.0 group) extent of the NAc-S viral infection in D1-Cre rats. Location of fiber-optic cannulas in the NAc-S (gray for eYFP group and blue for eNpHR3.0 group) is also represented. Distances are indicated in mm from bregma. (B) Micrographs showing viral expression in NAc-S D1-SPNs in the eYFP group (left) and eNpHR3.0 group (right). (C) During Pavlovian conditioning, magazine entry rates were similar across group (Group – eYFP vs. eNpHR3.0: p = 0.13) and increased across days (Day: F1,18 = 88.24, p < 0.001, η2=0.83) regardless of group (Group x Day: p = 0.45). The rates were higher in the presence of the stimuli than in their absence (Period, S vs. pre: F1,18 = 259.04, p < 0.001, η2 = 0.94) irrespective of group (Group x Period: p = 0.12). The discrimination between the two periods increased as training progressed (Day Period: F1,18 = 66.84, p < 0.001, η2 = 0.79) regardless of group (Group x Day x Period: p = 0.72). (D) During instrumental conditioning, lever press rates were similar across group (Group: p = 0.16) and increased as training progressed (Day: F1,18 = 190.83, p < 0.001, η2 = 0.91). The increase was more pronounced in the eNpHR3.0 group than in the eYFP group (Group x Day: F1,18 = 5.40, p < 0.05, η2 = 0.23). (E) Raw data for lever presses during the Pavlovian instrumental transfer (PIT) test. (F) During the PIT test, magazine entry rates were higher in the eNpHR3.0 group than the eYFP group (Group: F1,18 = 7.84, p < 0.05, η2 = 0.30). The rates remained unaffected by LED activation (LED – ON vs. OFF, p = 0.16) regardless of group (Group x LED: p = 0.89). The rates were higher in the presence of the stimuli than in their absence (Period: F1,18 = 26.58, p < 0.001, η2 = 0.60) irrespective of group (Group x Period: p = 0.12). The discrimination between the two periods was unaffected by LED activation (Period x LED: p = 0.15) irrespective of group (Group x Period x LED: p = 0.78). (G) During the choice test, lever press rates were similar across groups (Group: p = 0.15) and were higher on the lever earning the valued outcome (Lever – Valued vs. Devalued: F1,18 = 49.03, p < 0.001, η2 = 0.74) irrespective of group (Group x Lever: p = 0.33). Lever press rates were unaffected by LED activation (LED: p = 0.61) irrespective of group (Group x LED: p = 0.06). The difference between rates on the valued and devalued lever was left intact by LED activation (Lever x LED: p = 0.26) but this depended on the group considered (Group x Lever x LED: F1,18 = 10.04, p < 0.01, η2 = 0.36). Nevertheless, simple effect analyses revealed that lever press rates were higher on the valued lever compared to the devalued lever in both groups whether the LED was OFF or ON (eYFP-OFF: F1,9 = 13.29, p < 0.01, η2 = 0.57; eYFP-ON: F1,9 = 47.45, p < 0.001, η2 = 0.84; eNpHR3.0-OFF: F1,9 = 31.68, p < 0.001, η2 = 0.78; eNpHR3.0-ON: F1,9 = 6.36, p < 0.05, η2 = 0.41). Data are shown as mean ± SEM. Panels E–G include individual data points for female (filled circle) and male (open circle) rats. Asterisks denote significant effect (***p < 0.001; *p < 0.05), n.s., nonsignificant.
Figure 4 with 1 supplement
NAc-S D2-SPNs mediate for outcome-specific Pavlovian instrumental transfer (PIT).
(A) A2a-Cre rats were bilaterally infused in the NAc-S with DIO-eYFP (black; 4 females and 4 males) or DIO-eNpHR3.0 (red; 4 females and 2 males). Fiber-optic cannulas were implanted above the NAc-S to provide 625 nm LED illumination (continuous). (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 625 nm 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).
Figure 4—figure supplement 1
Histological and behavioral data related to Figure 4.
A2a-Cre rats were bilaterally infused in the NAc-S with DIO-eYFP (black; 4 females and 4 males) or DIO-eNpHR3.0 (red; 4 females and 2 males). (A) Minimal (light gray for eYFP group and light red for eNpHR3.0 group) and maximal (darker gray for eYFP group and darker red for eNpHR3.0 group) extent of the NAc-S viral infection in A2a-Cre rats. Location of fiber-optic cannulas in the NAc-S (gray for eYFP group and red for eNpHR3.0 group) is also represented. Distances are indicated in mm from bregma. (B) Micrographs showing viral expression in NAc-S D2-SPNs in the eYFP group (left) and eNpHR3.0 group (right). (C) During Pavlovian conditioning, magazine entry rates were similar across group (Group: p = 0.26) and increased across days (Day: F1,12 = 89.63, p < 0.001, η2 = 0.88) regardless of group (Group x Day: p = 0.85). The rates were higher in the presence of the stimuli than in their absence (Period: F1,12 = 160.90, p < 0.001, η2 = 0.93) irrespective of group (Group x Period: p = 0.39). The discrimination between the two periods increased as training progressed (Day x Period: F1,12 = 74.66, p < 0.001, η2 = 0.86) regardless of group (Group x Day x Period: p = 0.70). (D) During instrumental conditioning, lever press rates were similar across group (Group: p = 0.45) and increased as training progressed (Day: F1,12 = 116.85, p < 0.001, η2 = 0.91) irrespective of group (Group x Day: p = 0.39). (E) Raw data for lever presses during the Pavlovian instrumental transfer (PIT) test. (F) During the PIT test, magazine entry rates were similar across groups (Group: p = 0.053). The rates remained unaffected by LED activation (LED: p = 0.055) regardless of group (Group x LED: p = 0.44). The rates were higher in the presence of the stimuli than in their absence (Period: F1,12 = 50.37, p < 0.001, η2 = 0.81) irrespective of group (Group x Period: p = 0.09). The discrimination between the two periods was unaffected by LED activation (Period x LED: p = 0.89) irrespective of group (Group x Period x LED: p = 0.75). (G) During the choice test, lever press rates were similar across groups (Group: p = 0.86) and were higher on the lever earning the valued outcome (Lever: F1,12 = 66.95, p < 0.001, η2 = 0.85) irrespective of group (Group x Lever: p = 0.54). Lever press rates were unaffected by LED activation (LED: p = 0.98) irrespective of group (Group x LED: p = 0.85). The difference between rates on the valued and devalued lever was left intact by LED activation (Lever x LED: p = 0.71) regardless of group (Group x Lever x LED: p = 0.36). Data are shown as mean ± SEM. Panels E–G include individual data points for female (filled circle) and male (open circle) rats.
Figure 5 with 1 supplement
NAc-S D1-SPNs projections to the ventral pallidum (VP) mediate outcome-specific Pavlovian instrumental transfer (PIT).
(A) D1-Cre rats were bilaterally infused in the NAc-S with DIO-eYFP (black; 3 females and 5 males) or DIO-eNpHR3.0 (blue; 6 females and 6 males). Fiber-optic cannulas were implanted above the VP to provide 625 nm LED illumination (continuous). (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 625 nm 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).
Figure 5—figure supplement 1
Histological and behavioral data related to Figure 5.
D1-Cre rats were bilaterally infused in the NAc-S with DIO-eYFP (black; 3 females and 5 males) or DIO-eNpHR3.0 (blue; 6 females and 6 males). (A) Minimal (light gray for eYFP group and light blue for eNpHR3.0 group) and maximal (darker gray for eYFP group and darker blue for eNpHR3.0 group) extent of the NAc-S viral infection in D1-Cre rats. Location of fiber-optic cannulas in the ventral pallidum (VP) (gray for eYFP group and blue for eNpHR3.0 group) is also represented. Distances are indicated in mm from bregma. (B) Micrographs showing viral expression in NAc-S D1-SPNs and their terminals in the VP for the eYFP group (left) and eNpHR3.0 group (right). (C) During Pavlovian conditioning, magazine entry rates were similar across group (Group: p = 0.79) and increased across days (Day: F1,18 = 83.14, p < 0.001, η2 = 0.82) regardless of group (Group x Day: p = 0.50). The rates were higher in the presence of the stimuli than in their absence (Period: F1,18 = 129.21, p < 0.001, η2 = 0.88) irrespective of group (Group x Period: p = 0.31). The discrimination between the two periods increased as training progressed (Day x Period: F1,18 = 48.18, p < 0.001, η2 = 0.73) regardless of group (Group x Day x Period: p = 0.49). (D) During instrumental conditioning, lever press rates were similar across groups (Group: p = 0.90) and increased as training progressed (Day: F1,18 = 171.16, p < 0.001, η2 = 0.90) irrespective of group (Group x Day: p = 0.65). (E) Raw data for lever presses during the Pavlovian instrumental transfer (PIT) test. (F) During the PIT test, magazine entry rates were similar across group (Group: p = 0.72). The rates remained unaffected by LED activation (LED: p = 0.77) regardless of group (Group x LED: p = 0.96). The rates were higher in the presence of the stimuli than in their absence (Period: F1,18 = 20.80, p < 0.001, η2 = 0.54) irrespective of group (Group x Period: p = 0.86). The discrimination between the two periods was unaffected by LED activation (Period x LED: p = 0.73) irrespective of group (Group x Period x LED: p = 0.56). (G) During the choice test, lever press rates were similar across group (Group: p = 0.72) and were higher on the lever earning the valued outcome (Lever: F1,18 = 36.12, p < 0.001, η2 = 0.67) irrespective of group (Group x Lever: p = 0.97). Lever press rates were unaffected by LED activation (LED: p = 0.89) irrespective of group (Group x LED: p = 0.45). The difference between rates on the valued and devalued lever was left intact by LED activation (Lever x LED: p = 0.99) regardless of group (Group x Lever x LED: p = 0.27). Data are shown as mean ± SEM. Panels E–G include individual data points for female (filled circle) and male (open circle) rats.
Figure 6 with 1 supplement
NAc-S D2-SPNs projections to the ventral pallidum (VP) do not mediate outcome-specific Pavlovian instrumental transfer (PIT).
(A) A2a-Cre rats were bilaterally infused in the NAc-S with DIO-eYFP (black; 3 females and 5 males) or DIO-eNpHR3.0 (red; 2 females and 5 males). Fiber-optic cannulas were implanted above the VP to provide 625 nm LED illumination (continuous). (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 625 nm 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.
Figure 6—figure supplement 1
Histological and behavioral data related to Figure 6.
A2a-Cre rats were bilaterally infused in the NAc-S with DIO-eYFP (black; 3 females and 5 males) or DIO-eNpHR3.0 (red; 2 females and 5 males). (A) Minimal (light gray for eYFP group and light red for eNpHR3.0 group) and maximal (darker gray for eYFP group and darker red for eNpHR3.0 group) extent of the NAc-S viral infection in A2a-Cre rats. Location of fiber-optic cannulas in the ventral pallidum (VP) (gray for eYFP group and red for eNpHR3.0 group) is also represented. Distances are indicated in mm from bregma. (B) Micrographs showing viral expression in NAc-S D2-SPNs and their terminals in the VP for the eYFP group (left) and eNpHR3.0 group (right). (C) During Pavlovian conditioning, magazine entry rates were similar across group (Group: p = 0.83) and increased across days (Day: F1,13 = 23.82, p < 0.001, η2 = 0.65) regardless of group (Group x Day: p = 0.42). The rates were higher in the presence of the stimuli than in their absence (Period: F1,13 = 767.22, p < 0.001, η2 = 0.98) irrespective of group (Group x Period: p = 0.64). The discrimination between the two periods increased as training progressed (Day x Period: F1,13 = 106.62, p < 0.001, η2 = 0.89) regardless of group (Group x Day x Period: p = 0.40). (D) During instrumental conditioning, lever press rates were higher in the eYFP group than the eNpHR3.0 group (Group: F1,13 = 6.18, p < 0.03, η2 = 0.32). The rates increased as training progressed (Day: F1,13 = 155.04, p < 0.001, η2 = 0.92) irrespective of group (Group x Day: p = 0.10). (E) Raw data for lever presses during the Pavlovian instrumental transfer (PIT) test. (F) During the PIT test, magazine entry rates were similar across groups (Group: p = 0.09). Larger rates were observed with LED activation (LED: F1,13 = 6.26, p < 0.05, η2 = 0.32) but this was true regardless of group (Group x LED: p = 0.74). The rates were higher in the presence of the stimuli than in their absence (Period: F1,13 = 76.18, p < 0.001, η2 = 0.86), especially in the eNpHR3.0 group compared to the eYFP group (Group x Period: F1,13 = 6.97, p < 0.05, η2 = 0.35). The discrimination between the two periods was increased by LED activation (Period x LED: F1,13 = 11.17, p < 0.01, η2 = 0.46) but this was true regardless of group (Group x Period x LED: p = 0.26). (G) During the choice test, lever press rates were similar across groups (Group: p = 0.99) and were higher on the lever earning the valued outcome (Lever: F1,13 = 27.04, p < 0.001, η2 = 0.68) irrespective of group (Group x Lever: p = 0.97). Lever press rates were unaffected by LED activation (LED: p = 0.67) irrespective of group (Group x LED: p = 0.51). The difference between rates on the valued and devalued lever was left intact by LED activation (Lever x LED: p = 0.62) regardless of group (Group x Lever x LED: p = 0.53). Data are shown as mean ± SEM. Panels E–G include individual data points for female (filled circle) and male (open circle) rats.
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The influence of nucleus accumbens shell D1 and D2 neurons on outcome-specific Pavlovian instrumental transfer
eLife 14:RP107566.
https://doi.org/10.7554/eLife.107566.4
