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. 2019 Mar 6;5(3):eaav1640. doi: 10.1126/sciadv.aav1640

Fig. 3. DBBGABA neurons mediate the effect of LHGABA→DBB projections.

Fig. 3

(A to C) DBBGABA neurons receive direct synaptic input from LHGABA neurons. (A) Schematic showing Vgat-cre mice receiving LH injection of AAV-Flex-ChR2 (LHVgat-ChR2) and DBB injection of AAV-DIO-hM3Dq-mCherry and implant of optic fiber (DBBVgat-hM3Dq), and Vglut2-cre mice receiving LHVglut2-Fas-ChR2:DBBhM3Dq:fiber. (B and C) Brain slice electrophysiology of oIPSC in DBBmCherry+ and DBBmCherry− neurons of LHVgat-ChR2:DBBVgat-hM3Dq-mCherry mice (B; n = 3) and LHVglut2-Fas-ChR2:DBBVglut2-hM3Dq-mCherry mice (C; n = 2). (D and E) Effect of CNO-mediated activation of DBBhM3Dq on photostimulation-induced feeding in LHVgat-ChR2:DBBVgat-hm3Dq mice [one-way analysis of variance (ANOVA) and Tukey post hoc test] (D; n = 11; saline:photostimulation, P < 0.001; saline:CNO-photostimulation, P = 0.07: photostimulation:CNO-photostimulation, P < 0.001), and LHVglut2-Fas-ChR2:DBBVglut2-hm3Dq mice) (E; n = 8; saline:photostimulation, P = 0.002; saline:CNO-photostimulation, P = 0.006; photostimulation:CNO-photostimulation, P = 0.88). (F) Simultaneous recording of LHVgat-GCaMP6m and DBBVgat-GCaMP6m population activity during feeding bouts (shaded areas). (G and H) Effect of photostimulation of LHVgat-ChR2 cell bodies on DBBVgat-GCaMP6 neuronal activity at various frequencies (G) and pulse durations (H). *P < 0.05; **P < 0.01; ***P < 0.001.