biostudies-literatureYang MNICHD NIH HHSHHS | NIH | Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD)HHS | NIH | National Institute of General Medical Sciences (NIGMS)e2508783122https://www.ebi.ac.uk/biostudies/studies/S-EPMC12452909biostudies-literatureUnknown122(37)Ovulation is an intricate process that is essential for reproductive success. In <i>Drosophila melanogaster</i>, ovulation increases after mating. This increase is initiated by the male seminal fluid protein ovulin and is executed by female pathways, including octopamine (OA) neuronal signaling. Despite OA signaling's central role in ovulation regulation, the broader molecular landscape underlying female control of ovulation remains poorly understood. Here, using ovulin as a probe, we performed evolutionary rate covariation and AlphaFold-Multimer prediction screens to identify candidate female ovulation-regulating proteins. Ovulation assays performed on knockdowns or mutants of identified membrane-protein candidates revealed seven important female ovulation regulators: Lgr3, GabaβR1, SIFaRProceedings of the National Academy of Sciences of the United States of AmericaFemale membrane proteins regulate postmating ovulation in &lt;i&gt;Drosophila melanogaster&lt;/i&gt; by ovulin-dependent and -independent pathways.PMC12452909R37 HD038921R37-HD038921F32-GM155980R35-GM136258Apger-McGlaughon JClark NLFindlay GDVignogna RCWolfner MFChoi JYYang MWhite MAFromme JCfalseFemale membrane proteins regulate postmating ovulation in &lt;i&gt;Drosophila melanogaster&lt;/i&gt; by ovulin-dependent and -independent pathways.Ovulation is an intricate process that is essential for reproductive success. In <i>Drosophila melanogaster</i>, ovulation increases after mating. This increase is initiated by the male seminal fluid protein ovulin and is executed by female pathways, including octopamine (OA) neuronal signaling. Despite OA signaling's central role in ovulation regulation, the broader molecular landscape underlying female control of ovulation remains poorly understood. Here, using ovulin as a probe, we performed evolutionary rate covariation and AlphaFold-Multimer prediction screens to identify candidate female ovulation-regulating proteins. Ovulation assays performed on knockdowns or mutants of identified membrane-protein candidates revealed seven important female ovulation regulators: Lgr3, GabaβR1, SIFaR2025-01-01T00:00:00Z2025 Sep2026-06-04T10:10:09.999Z2026-05-08T03:10:10.54ZS-EPMC124529094092092110.1073/pnas.2508783122