(A) At day 10 of pregnancy chronic kisspeptin-234 (KP-234) administration had no significant effect on glucose tolerance; however, at day 16 of pregnancy mice treated with KP-234 had significantly impaired glucose tolerance following i.p. glucose administration (2 g/kg) when compared with untreated pregnant controls, as determined by both comparison of individual time points (2-way repeated-measures ANOVA; 60 minutes P = 0.022; 90 minutes P = 0.046) and (B) glucose AUC over the course of the test (1-way ANOVA, P = 0.031). (C) Blocking endogenous kisspeptin signaling with KP-234 did not have any effect on fasting plasma insulin levels at either day 10 or day 16 of pregnancy. At day 16 chronic KP-234 treatment resulted in significantly reduced insulin release in response to i.p. glucose administration (2 g/kg) after 30 minutes when compared with pregnant controls (1-way ANOVA, P = 0.009). A similar trend was observed at day 10; however, this was not significant. (D) KP-234 had no effect on the plasma glucose response to i.p. insulin administration (0.75 IU/kg) when compared to untreated controls at either day 12 or day 18 or pregnancy, through both comparison of individual time points and (E) glucose AUC. Mean ± SEM; n = 6; *P < 0.05 vs. control.