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Open Access Research article

Identification of ejaculated proteins in the house mouse (Mus domesticus) via isotopic labeling

Matthew D Dean12*, Geoffrey D Findlay3, Michael R Hoopmann3, Christine C Wu4, Michael J MacCoss3, Willie J Swanson3 and Michael W Nachman2

Author Affiliations

1 Molecular and Computational Biology, University of Southern California, 1050 Childs Way, Los Angeles, CA, USA

2 Ecology and Evolutionary Biology, University of Arizona, Tucson, AZ, USA

3 Department of Genome Sciences, University of Washington, Seattle, WA, USA

4 Department of Cell Biology, University of Pittsburgh, Pittsburgh, PA, USA

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BMC Genomics 2011, 12:306  doi:10.1186/1471-2164-12-306

Published: 10 June 2011

Abstract

Background

Seminal fluid plays an important role in successful fertilization, but knowledge of the full suite of proteins transferred from males to females during copulation is incomplete. The list of ejaculated proteins remains particularly scant in one of the best-studied mammalian systems, the house mouse (Mus domesticus), where artificial ejaculation techniques have proven inadequate. Here we investigate an alternative method for identifying ejaculated proteins, by isotopically labeling females with 15N and then mating them to unlabeled, vasectomized males. Proteins were then isolated from mated females and identified using mass spectrometry. In addition to gaining insights into possible functions and fates of ejaculated proteins, our study serves as proof of concept that isotopic labeling is a powerful means to study reproductive proteins.

Results

We identified 69 male-derived proteins from the female reproductive tract following copulation. More than a third of all spectra detected mapped to just seven genes known to be structurally important in the formation of the copulatory plug, a hard coagulum that forms shortly after mating. Seminal fluid is significantly enriched for proteins that function in protection from oxidative stress and endopeptidase inhibition. Females, on the other hand, produce endopeptidases in response to mating. The 69 ejaculated proteins evolve significantly more rapidly than other proteins that we previously identified directly from dissection of the male reproductive tract.

Conclusion

Our study attempts to comprehensively identify the proteins transferred from males to females during mating, expanding the application of isotopic labeling to mammalian reproductive genomics. This technique opens the way to the targeted monitoring of the fate of ejaculated proteins as they incubate in the female reproductive tract.

Keywords:
seminal fluid; ejaculate; evolution