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Type of publication: Journal Article
Type of document: Full Paper

Year: 2015

Authors: Artemenko, K; Horáková, J; Steinberger, B; Besenfelder, U; Brem, G; Bergquist, J; Mayrhofer, C

Title: A proteomic approach to monitor the dynamic response of the female oviductal epithelial cell surface to male gametes.

Source: J Proteomics. 2015; 113:1-14

Authors Vetmeduni Vienna:

Besenfelder Urban
Brem Gottfried
Itze-Mayrhofer Corina
Steinberger Birgit

Vetmed Research Units
Institute of Animal Breeding and Genetics, Unit of Reproductive Biology

Project(s): Identification of molecules that effect spermatozoa and fertilization success

Sophisticated strategies to analyze cell surface proteins are indispensable to study fundamental biological processes, such as the response of cells to environmental changes or cell-cell communication. Herein, we describe a refined mass spectrometry-based approach for the specific characterization and quantitation of cell surface proteins expressed in the female reproductive tract. The strategy is based on in situ biotinylation of rabbit oviducts, affinity enrichment of surface exposed biotin tagged proteins and dimethyl labeling of the obtained tryptic peptides followed by LC-MS/MS analysis. This approach proved to be sensitive enough to analyze small sample amounts (<1μg) and allowed further to trace the dynamic composition of the surface proteome of the oviductal epithelium in response to male gametes. The relative protein expression ratios of 175 proteins were quantified. Thirty-one of them were found to be altered over time, namely immediately, 1h and 2h after insemination compared to the time-matched control groups. Functional analysis demonstrated that structural reorganization of the oviductal epithelial cell surface was involved in the early response of the female organ to semen. In summary, this study outlines a workflow that is capable to monitor alterations in the female oviduct that are related to key reproductive processes in vivo.

Keywords Pubmed: Animals
Cell Communication/physiology*
Epithelial Cells/cytology
Epithelial Cells/metabolism*
Fallopian Tubes/cytology
Fallopian Tubes/metabolism*
Membrane Proteins/biosynthesis*

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