Figure 3.

Double-label ISH for cfos colocalization in Kiss1 neurons during the time of the LH surge. (A) Representative photomicrographs of cfos (silver grains) colocalizing with AVPV/PeN Kiss1 neurons (red fluorescence) in E₂-treated males and females at two time points. Kiss1 neurons in female mice euthanized at CT12 have substantially more cfos colocalization than at CT4; male Kiss1 neurons have low cfos induction at both time points. Green arrowheads, example Kiss1 cells with cfos; blue arrows, example Kiss1 cells lacking cfos expression. (B) Quantification of the percent colocalization of cfos in AVPV/PeN Kiss1 neurons in each sex across the circadian day. Different lowercase letters indicate significantly different groups (P < 0.05).