Figure 7. Functional assessment of Kiss1 neurons in Kiss1^Cre:Nhlh2^fl/fl male mice under fed and fasting conditions.

Central (icv) administration of Kp-10 (50 pmol/mouse in 5 μl) (Nhlh2^fl/fl [n=10] and Kiss1^Cre:Nhlh2^fl/fl [n=7]) (A) or senktide (600 pmol/mouse in 5 μl) Nhlh2^fl/fl (n=9) and Kiss1^Cre:Nhlh2^fl/fl (n=7) (B) was performed in adult male Kiss1^Cre:Nhlh2^fl/fl mice and controls. Blood samples were collected before and 30 min after treatment. Groups with different letters are significantly different two-way ANOVA followed by Bonferroni test. In (C), the ability of Kiss1^Cre:Nhlh2^fl/fl male mice to generate a compensatory LH rise after gonadectomy was assessed before surgery and 10 days post-gonadectomy. Nhlh2^fl/fl (n=10) and Kiss1^Cre:Nhlh2^fl/fl (n=7). Two-way ANOVA followed by Bonferroni test. (D) Response of LH in gonadectomized male mice during 24 hr of fasting and 12 hr of refeeding (Nhlh2^fl/fl [n=10] and Kiss1^Cre:Nhlh2^fl/fl [n=7]); *p<0.05 compared with the control group at 12 hr. Two-way ANOVA followed by Fisher’s test. The area under the curve (AUC) was determined during fasting and refeeding periods. (E) Response of mice to overnight fasting followed by leptin administration. Blood samples were collected before and 30 min after leptin (2 μg/mouse in 5 μl) treatment. Nhlh2^fl/fl (n=9) and Kiss1^Cre:Nhlh2^fl/fl (n=6); Groups with different letters are significantly different. Two-way ANOVA followed by Bonferroni post hoc test. Data presented as median (dotted line)+distribution of the data and its probability density (violin plot) or the mean± SEM (in (D)). cKO=Kiss1^Cre:Nhlh2^fl/fl. LH, luteinizing hormone.