(A) Schematic showing experimental protocol in which GnRH-Cre mice are injected with Cre-dependent Flex-hM4D(Gi)-mCherry AAVs bilaterally into the region of the median eminence and, later, CNO injected into the rostral preoptic area (rPOA). MS, medial septum. CNO, Clozapine N-oxide. (B,C) LH surge profiles in all GnRH-Cre OVX+E+P female mice following rPOA injection of saline (C, n = 9) or CNO (D, n = 10). (D) Mean (± SEM) LH levels. **p<0.01, ***p<0.001, two-way repeated measures ANOVA with Holm-Sidak test. (E-H), Representative profiles of LH pulsatile secretion in GnRH-Cre OVX female mice with rPOA injection of saline (E, F) or CNO (G, H). LH pulses are indicated by asterisks. Mean (± SEM) LH pulse amplitude (I) and frequency (J) in GnRH-cre OVX mice with rPOA injection of saline (n = 8) and CNO (n = 9).