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. 2023 Feb 2;26(3):106123. doi: 10.1016/j.isci.2023.106123

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© 2023 The Authors

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

PMC Copyright notice

The active states of SPN reflect sexual receptivity

(A) Representative images of GCaMP6 signals (green) in SPN of a virgin (A1-2) and a newly mated (A3-4) female. Red signals in (A2) and (A4) show the corresponding cell clusters. The genotype is SPNsplit-Gal4>UAS-GCaMP6, UAS-myr::RFP. The dashed lines indicate the profiles of brains.

(B–D) Normalized fluorescence intensities of GCaMP6 signals in SPN of virgin females (gray) and mated females (red) at different times after mating. (B) Comparison of the age-matched virgins and newly-mated females (n = 25, 26). (C) Comparison of the age-matched virgins and mated females 1 day after mating (n = 24, 24). (D) Comparison of normalized fluorescence intensities of GCaMP6 signals in SPN in three classes of females. The top panel shows the intensity values of three groups (n = 21, 20, 20). Group 1: virgin females (un-mated); Group 2: females mated with Crz-Gal4>UAS-Kir2.1 males; Group 3: females mated with wild-type males (Canton S). The bottom panel shows the confidence intervals for the difference between indicated groups (95% confidence).

(E) Representative images of SPN in virgin females responding to 60 μM SP treatment. (E1-2): before treatment. (E3-4): after SP treatment. Green indicates the GCaMP signals, and red labels the corresponding cell clusters. The genotype is SPNsplit-Gal4>UAS-GCaMP6, UAS-myr::RFP. The dashed lines indicate the profiles of brains.

(F) Change of normalized fluorescence intensity (ΔF/F) of GCaMP6 signals in SPN of virgin females, after SP treatment (n = 26). The light gray and red dots indicate the mean of each dataset.

All genotypes and experimental conditions are indicated with the plots. In the box-and-whisker plot, the whiskers mark the minimum and maximum, the box includes the 25th to 75th percentiles, and the line within the box indicates the median of the dataset. The unpaired t-test was performed for (B)-(C). The Kruskal–Wallis test was performed for (D). The paired t-test was performed for (F). ns, not significant (p> 0.05); ∗∗p< 0.01; ∗∗∗p< 0.001; ∗∗∗∗p< 0.0001. Scale bar in (A) and (E): 100 μm.