Fig. 7: SF1-positive cells in the VMHd are important for hemoglobin-dependent digging enhancement in lactating female mice.

a Schematic illustration of the animal setup and timeline for pharmacogenetic inhibition of SF1-expressing neurons in the VMHd. AAV-DIO-hM4Di-mCherry is injected into SF1-positive cells in the VMHd. Image adapted from Allen Mouse Brain Atlas⁴⁸. b A representative coronal section showing DREADD-Gi expression (mCherry-positive cells shown in red) in the VMHd. n = 8. Scale bar, 500 µm. c Quantification of the total digging duration (sec) of hemoglobin (Hb)-stimulated SF1-Cre lactating mothers with i.p. injection of saline or CNO. n = 4 for CNO group. n = 4 for saline group. Error bars, S.E.M. p = 0.030 by the two-sided Wilcoxon signed-rank test. d Schematic image and illustration of the setup and timeline for optogenetic activation of SF1-expressing neurons in the VMHd of lactating females. AAV-DIO-ChR2 or AAV-DIO-GFP are injected into SF1-positive cells in the VMHd and optic fibers are implanted above the target region. Image adapted from Allen Mouse Brain Atlas⁴⁸. e A representative coronal section showing ChR2 expression (eYFP-positive cells shown in green) in the VMHd. n = 5. Scale bar, 500 µm. f, g Quantification of digging behavior, with or without weak light stimulation (f 0 mW, g 0.01 mW). Error bars, S.E.M. p = 0.043 by the two-sided Wilcoxon signed-rank test, n = 5 for GFP and ChR2 group.