University of Veterinary Medicine Vienna - Research portal
Type of publication:
Type of document:
Yu, H; Hackenbroch, L; Meyer, FRL; Reiser, J; Razzazi-Fazeli, E; Nöbauer, K; Besenfelder, U; Vogl, C; Brem, G; Mayrhofer, C
Identification of Rabbit Oviductal Fluid Proteins Involved in Pre-Fertilization Processes by Quantitative Proteomics.
Proteomics. 2019; 19(5):e1800319
Authors Vetmeduni Vienna:
Vetmed Research Units
Institute of Animal Breeding and Genetics, Unit of Molecular Genetics
Institute of Animal Breeding and Genetics, Unit of Reproductive Biology
Identification of molecules that effect spermatozoa and fertilization success
- Oviductal fluid (ODF) proteins modulate and support reproductive processes in the oviduct. In the present study, proteins involved in the biological events that precede fertilization have been identified in the rabbit ODF proteome, isolated from the ampulla and isthmus of the oviduct at different time points within 8 h after intrauterine insemination. A workflow is used that integrates lectin affinity capture with stable-isotope dimethyl labeling prior to nanoLC-MS/MS analysis. In total, over 400 ODF proteins, including 214 lectin enriched glycoproteins, are identified and quantified. Selected data are validated by Western blot analysis. Spatiotemporal alterations in the abundance of ODF proteins in response to insemination are detected by global analysis. A subset of 63 potentially biologically relevant ODF proteins is identified, including extracellular matrix components, chaperones, oxidoreductases, and immunity proteins. Functional enrichment analysis reveals an altered peptidase regulator activity upon insemination. In addition to protein identification and abundance changes, N-glycopeptide analysis further identifies 281 glycosites on 199 proteins. Taken together, these results show, for the first time, the evolving oviductal milieu early upon insemination. The identified proteins are likely those that modulate in vitro processes, including spermatozoa function.© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Chromatography, High Pressure Liquidmethods
Tandem Mass Spectrometrymethods