title: Retention of Ejaculate by Drosophila melanogaster Females Requires the Male-Derived Mating Plug Protein PEBme.
abstract: Within the mated reproductive tracts of females of many taxa, seminal fluid proteins (SFPs) coagulate into a structure known as the mating plug (MP). MPs have diverse roles, including preventing female remating, altering female receptivity postmating, and being necessary for mated females to successfully store sperm. The Drosophila melanogaster MP, which is maintained in the mated female for several hours postmating, is comprised of a posterior MP (PMP) that forms quickly after mating begins and an anterior MP (AMP) that forms later. The PMP is composed of seminal proteins from the ejaculatory bulb (EB) of the male reproductive tract. To examine the role of the PMP protein PEBme in D. melanogaster reproduction, we identified an EB GAL4 driver and used it to target PEBme for RNA interference (RNAi) knockdown. PEBme knockdown in males compromised PMP coagulation in their mates and resulted in a significant reduction in female fertility, adversely affecting postmating uterine conformation, sperm storage, mating refractoriness, egg laying, and progeny generation. These defects resulted from the inability of females to retain the ejaculate in their reproductive tracts after mating. The uncoagulated MP impaired uncoupling by the knockdown male, and when he ultimately uncoupled, the ejaculate was often pulled out of the female. Thus, PEBme and MP coagulation are required for optimal fertility in D. melanogaster. Given the importance of the PMP for fertility, we identified additional MP proteins by mass spectrometry and found fertility functions for two of them. Our results highlight the importance of the MP and the proteins that comprise it in reproduction and suggest that in Drosophila the PMP is required to retain the ejaculate within the female reproductive tract, ensuring the storage of sperm by mated females.
authors.affiliation: Department of Molecular Biology and Genetics, Cornell University, Ithaca, New York 14853 fwa5@cornell.edu mfw5@cornell.edu.
authors.forename: Frank W
authors.initials: FW
authors.lastname: Avila
authors.affiliation: Department of Molecular Biology and Genetics, Cornell University, Ithaca, New York 14853.
authors.forename: Allie B
authors.initials: AB
authors.lastname: Cohen
authors.affiliation: Department of Molecular Biology and Genetics, Cornell University, Ithaca, New York 14853.
authors.forename: Fatima S
authors.initials: FS
authors.lastname: Ameerudeen
authors.affiliation: Department of Entomology, Cornell University, Ithaca, New York 14853.
authors.forename: David
authors.initials: D
authors.lastname: Duneau
authors.affiliation: Department of Molecular Biology and Genetics, Cornell University, Ithaca, New York 14853.
authors.forename: Shruthi
authors.initials: S
authors.lastname: Suresh
authors.affiliation: Department of Molecular Biology and Genetics, Cornell University, Ithaca, New York 14853.
authors.forename: Alexandra L
authors.initials: AL
authors.lastname: Mattei
authors.affiliation: Department of Molecular Biology and Genetics, Cornell University, Ithaca, New York 14853 fwa5@cornell.edu mfw5@cornell.edu.
authors.forename: Mariana F
authors.identifier: http://orcid.org/0000-0003-2701-9505
authors.initials: MF
authors.lastname: Wolfner
mesh_terms: D000818:Animals; D029721:Drosophila Proteins / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000378:metabolism* / Q000502:physiology; D004542:Ejaculation; D005260:Female; D005298:Fertility; D055785:Gene Knockdown Techniques; D008297:Male; D034622:RNA Interference; D012726:Sexual Behavior, Animal; D013094:Spermatozoa / Q000378:metabolism; D014599:Uterus / Q000378:metabolism
chemical_list: D029721:Drosophila Proteins
keywords: Drosophila reproduction; PEBme; mating plug; sperm storage