Fig. 3: Overexpression of BDNF in vlOFC-to-DMS or M2-to-DLS neurons does not alter alcohol intake.

a–d Overexpression of BDNF in vlOFC-to-DMS projecting neurons. a Schematic representation of BDNF overexpression in vlOFC-to-DMS circuit. Mice received bilateral injections of AAV2-DIO-BDNF-mCherry or AAV2-DIO-mCherry in the vlOFC and AAVretro-Cre-GFP in the DMS. b Representative image depicting targeting of AAV2-DIO-BDNF-mCherry in vlOFC and AAVretro-Cre-GFP in the DMS. Red cells indicate the expression of Cre and infection of AAV2-DIO-BDNF-mCherry, confirming the overexpression of BDNF in vlOFC neurons projecting to the DMS. Top left panel depicts mCherry (red) and DAPI (cyan). Bottom left panel depicts GFP (green) and DAPI (cyan). Top and bottom right panels depict mCherry (red) and GFP (green). c Three weeks after the surgery, mice underwent IA20%2BC for 7 weeks in the home cage (Supplementary Table 1), and alcohol intake was recorded. d Alcohol preference was calculated as the ratio of alcohol intake relative to total fluid intake. e–h Overexpression of BDNF in M2-to-DLS projecting neurons. e Schematic representation of BDNF overexpression in M2-to-DLS circuit. Mice received bilateral injections of AAV2-DIO-BDNF-mCherry or AAV2-DIO-mCherry in M2 and AAVretro-Cre-GFP in the DLS. f Representative image depicting targeting of AAV2-DIO-BDNF-mCherry in M2 and AAVretro-Cre-GFP in the DLS. Red cells indicate expression of the Cre and infection with AAV2-DIO-BDNF-mCherry confirming the overexpression of BDNF in M2 neurons projecting to the DLS. Top left panel depicts mCherry (red) and DAPI (cyan). Bottom left panel depicts GFP (green) and DAPI (cyan). Top and bottom right panels depict mCherry (red) and GFP (green). g) Three weeks after the surgery, mice underwent IA20%2BC for 7 weeks in the home cage (Supplementary Table 1), and alcohol intake was recorded. h Alcohol preference was calculated as the ratio of alcohol intake relative to total fluid. Data are represented as mean ± SEM. n = 8–10 per group.