title: Seminal fluid protein depletion and replenishment in the fruit fly,Drosophila melanogaster: an ELISA-based method for tracking individual ejaculates.
abstract: In many species, seminal fluid proteins (SFPs) affect female post-mating behavioral patterns, including sperm storage, egg laying, feeding, and remating. Yet, few studies have investigated the patterns of allocation, depletion, and replenishment of SFPs in male animals, despite the importance of these proteins to male and female reproductive success. To investigate such SFP dynamics, it is necessary to have a sensitive method for quantifying SFP levels in males and mated females. We developed such a method by adapting the enzyme-linked immunosorbent assay (ELISA) using anti-SFP antibodies. Here, we first use twoDrosophila melanogasterSFPs (ovulin and sex peptide) to demonstrate that ELISAs provide accurate measures of SFP levels. We find that, consistent with previous data from Western blotting or immunofluorescence studies, levels of both ovulin and sex peptide decline in the mated female with time since mating, but they do so at different rates. We then use ELISAs to show that males become depleted of SFPs with repeated matings, but that previously mated males are able to transfer "virgin" levels of SFPs after 3 days of sexual inactivity. Finally, we demonstrate that ELISAs can detect SFPs from wild-caughtD. melanogastermales and, thus, potentially can be used to track mating patterns in the wild. This method of measuring SFP dynamics can be used in a wide range of species to address questions related to male reproductive investment, female mating history, and variation in female post-mating behavioral changes.
authors.affiliation: Department of Molecular Biology and Genetics, Cornell University, 421 Biotechnology Building, Ithaca, NY 14853, USA.
authors.forename: Laura K
authors.initials: LK
authors.lastname: Sirot
authors.affiliation: Department of Molecular Biology and Genetics, Cornell University, 421 Biotechnology Building, Ithaca, NY 14853, USA.
authors.forename: Norene A
authors.initials: NA
authors.lastname: Buehner
authors.affiliation: Biological Sciences Department, Binghamton University, Vestal Parkway East, P.O. Box 6000, Binghamton, NY 13902, USA.
authors.forename: Anthony C
authors.initials: AC
authors.lastname: Fiumera
authors.affiliation: Department of Molecular Biology and Genetics, Cornell University, 421 Biotechnology Building, Ithaca, NY 14853, USA.
authors.forename: Mariana F
authors.initials: MF
authors.lastname: Wolfner
keywords: Drosophila melanogaster; Ejaculate quantification; Ovulin; Reproductive investment; Sex peptide
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