Activation of AstC-Gal4 neurons inhibits both egg production and egg laying. (A) Number of eggs laid per female (n = 40 to 60) of the indicated neuropeptide-Gal4 (NP-Gal4) genotype crossed with UAS-dTrpA1 over 48 h at 30 °C (hence activation of dTrpA1). The means ± SEM of the data are shown. Dunnett’s test; *P < 0.05, **P < 0.005, ***P < 0.001 for comparisons against UAS-dTrpA1 alone (yellow bar). The red arrowhead highlights AstC-Gal4/UAS-dTrpA1. (B, Upper) Behavioral protocol for measuring mated female egg laying used in A and B. (Lower) Number of eggs laid per female (n = 40 to 60) of the indicated genotype over 48 h at the indicated temperature (30 °C, red and 21 °C, blue). The means ± SEM of the data are shown. One-way ANOVA followed by Tukey’s test for multiple comparisons; ***P < 0.001 for comparisons of AstC-Gal4/UAS-dTrpA1 against the appropriate parental (light red bars) and temperature controls (blue bars). (C, Top) Protocol for measuring virgin female egg production. (Middle) Number of mature eggs per virgin female (n = 40 to 60) of the indicated genotype kept for 3 d posteclosion at the indicated temperature (30 °C, red and 21 °C, blue). The means ± SEM of the data are shown. One-way ANOVA followed by Tukey’s test for multiple comparisons; ***P < 0.001 for comparisons of AstC-Gal4/UAS-dTrpA1 against the appropriate parental and temperature controls. (Bottom) Ovaries of 3-d-old virgin females of the indicated genotype kept for 3 d posteclosion at 30 °C. (Scale bar, 1 mm.) (D, Top) Protocol for measuring mated female oviposition. (Middle) Number of mature eggs per mated female (n = 40 to 60) of the indicated genotype 2 d postcopulation at the indicated temperature (30 °C and 21 °C). The means ± SEM of the data are shown. One-way ANOVA followed by Tukey’s test for multiple comparisons; ***P < 0.001 for comparisons of AstC-Gal4/UAS-dTrpA1 against the appropriate parental and temperature controls. (Bottom) ovaries of mated females of the indicated genotype kept at 30 °C for 2 d postcopulation. (Scale bar, 1 mm.)