Fig. 7
Effects of activating and ablating mPOA Esr1+ neurons on mount and pup retrieval. a–c Optogenetic activation of mPOA Esr1+ neurons elicited mount and pup retrieval. a AAVs encoding Cre-inducible ChR2 were injected unilaterally into the mPOA of Esr1Cre animals. b Representative images showing expression of ChR2 as indicated by mCherry signal in Esr1+ neurons. Scale bar represents 200 μm for images on the top and 20 μm for images at the bottom. c Percentage of trials that animals displayed mount or pup retrieval in mCherry and ChR2 animals before, during (blue bars), and after phasic photostimulation (12 mW, 10 ms, 40 Hz, 15 s). For mount, N = 8 ChR2 and 9 mCherry males and 10 ChR2 and 8 mCherry females. For retrieval, N = 9 ChR2 and 9 mCherry males and 10 ChR2 and 8 mCherry females. Paired t test. *p < 0.05, **p < 0.01. d–f Ablation of mPOA Esr1+ neurons abolished sex differences in mount and pup retrieval. d AAVs encoding Cre-inducible taCasp3 were injected bilaterally into the mPOA of Esr1Cre animals to ablate Esr1+ neurons. e Example images of Esr1+ immunostaining in green and Nissl as the counter staining in blue in control and experimental animals of both sexes with quantification of Esr1+ signals shown on the right. Two-way ANOVA followed by Bonferroni post tests showed: virus effect, F(1, 22) = 44.43, p < 0.0001, sex effect, F(1, 22) = 7.38, p < 0.05, interaction effect, F(1, 22) = 2.52, p = 0.127. N = 8 Casp3 and 5 control males and 6 Casp3 and 7 control females. Scale bar, 300 μm. f Sex differences in mounting behavior and pup retrieval were abolished in Esr1Cre animals injected with Casp3 virus. Mount, two-way ANOVA followed by Bonferroni post tests for mount count: virus effect, F(1, 29) = 17.28, p < 0.001, sex effect, F(1,29) = 4.79, p < 0.05, interaction effect, F(1, 29) = 5.74, p < 0.05. Retrieval, two tailed Fisher’s exact test, p < 0.001, Casp3 vs. control in females. N = 9 Casp3 and 7 control males and 9 Casp3 and 8 control females
