Figure 4. SAG-1 neurons specifically require Dsx for a suite of female virgin behaviors.

(A and B) Suppression of Dsx within the SAG-1 lineage enhances egg-laying and compromises virgin receptivity, similar to whole-animal mir-iab-4/8 deletion and hth[BSmut] mutants. dsx-RNAi[JF] has a stronger effect.

(C) Strategy for temporal depletion of dsx. EL, egg-laying; Rec., receptivity.

(D and E) Analysis of egg-laying (D) and receptivity (E) in flies with developmental or adult knockdown of dsx, highlighting the developmental role of dsx for virgin adult behaviors.

(F and G) Modulation of Dsx levels also decreases vaginal plates opening (F) and induces ovipositor extrusions (G), comparable with mir-iab-4/8 deletion and hth [BSmut] mutants.

(H) Model for the genetic and spatial control of female virgin behavior in the VNC. Upper left, larval VNC illustrates the protein domains of Hth (in thoracic segments and A1) and Dsx (in A2–A9), and RNA domains of mir-iab-4 (A2–A7) and mir-iab-8 (A8–A9). Restricted neural populations that govern the female post-mating switch are distributed within A2–A9 (VT-switch neurons), including four SAG-1 neurons within the mir-iab-8 domain. In wild-type abdominal VNC, the activity of mir-iab-4/8 binding sites on hth-30 UTR, or depleted of dsx within VT-switch neurons (and in as few as those of highly restricted SAG-1 neurons) exhibit a switch from virgin to post-mated behaviors.

Mann-Whitney non-parametric test (A, F, and G); Fisher’s exact test (B and E). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; ns, not significant. Error bars, SEM. Eggs were collected over 3 days for all virgin genotypes and over the first 24 h after copulation in mated flies. V, virgin; M, mated.