The sterility phenotype of the octβ2r mutant female is due to defective ovulation.

(A) Copulation behavior. Virgin wild-type CS or octβ2r females were singly paired with naïve CS males to examine copulation behavior. The CS males paired with octβ2r females exhibited copulation latency and duration comparable to those paired with CS females (p>0.05 by Student’s t-test; n = 17–24). (B) Sperm retention. CS and octβ2r females were mated with dj-GFP males carrying GFP-tagged sperm (green) and the reproductive systems were dissected and counterstained with DAPI (blue). CS and octβ2r females had comparable sperm storage in the seminal receptacle and spermathecae at 24 and 48 h after mating. Spt, spermathecae; SmRcp, seminal receptacle. Scale bar, 50 µm. (C) Ovulation. CS and octβ2r females were mated with CS males and the reproductive systems were dissected to examine the location of the egg in the oviduct or uterus. The octβ2r females had significantly lower levels of ovulation than CS females (***, p<0.0001 by Student’s t-tests; n = 10). (D) Courtship behavior. Virgin CS and octβ2r females were singly paired with naïve CS males to measure courtship activity. The mated females were tested again with new naïve males 48 h later to measure courtship receptivity. The percentage of time that the CS males spent courting CS or octβ2r females represents courtship index. With both CS and octβ2r females, courtship activities of the males paired with mated females were significant lower than those of the males paired with virgin females (p<0.0001 by Mann-Whitney; n = 32–69). Thus, CS and octβ2r females have comparable pre- and post-mating courtship activity. ns, not significant.