Fig. 4. Projecting sites of GE- and GI-ERα^vlVMH neurons.

a Schematic experimental strategy using the Ad-iN/WED virus as a transsynaptic anterograde tracer to identify downstream circuits of female ERα^vlVMH neurons. b Immunoreactivity of WGA in the mpARH. The right panel shows the higher magnification image of the black box in the left panel. Scale bars are indicated in each panel. The similar results were replicated three times. c Schematic experimental strategy for recordings in WGA-labeled neurons in response to photostimulation of ChR2-labeled ERα^vlVMH-originated fibers within the mpARH in female mice. d Representative traces for light-evoked EPSCs, which were blocked by 30 μM D-AP5 and 50 μM CNQX, but not affected by 400 μM 4-AP and 1 μM TTX. e The percentage of WGA (+) neurons in the mpARH that showed light-evoked EPSCs or no response. f Amplitude of evoked EPSCs. g Latency of evoked EPSCs. N = 10 neurons from two mice per group. Results are shown as mean ± SEM. ***P = 0.0009 vs. other groups in one-way ANOVA followed by post hoc Sidak tests. h Schematic experimental strategy using the CAV2-Cre virus as a retrograde tracer to map GI-ERα^vlVMH and GE-ERα^vlVMH-originated neural circuits. i Composition of GI and GE neurons in the female ERα^vlVMH populations with specific projections to the mpARH, the LH, the DRN, and the MPB. N = 14–23 neurons from three mice per projecting site. Source data are provided as a Source Data Fig. 4.