Funding tool: Basic research projects
Project-ID: LS09-008
Project start: 02. August 2010
Project end: will follow
Runtime: 36 months / finished
Funding amount: € 219.700,00

Brief summary:

Reproductive biology is of great interest, not only for human medicine but also for animal breeding. The processes leading to fertilization and the establishment of pregnancy are highly complex. A major requisite for successful fertilization is the proper dialog between oviductal epithelial cells and spermatozoa. Independent of the hormonal status alterations of oviductal epithelial cells as well as of male gametes upon physiological interaction have been demonstrated.

However, our knowledge of the molecular basis of the response of epithelial cells to spermatozoa in vivo and the further activated or deactivated signalling processes allowing the establishment of an optimal environment for fertilization is still fragmentary.
In the present study we aim to analyse the spermatozoa induced signalling processes in epithelial cells on the proteome level in a time dependent manner using an in vivo animal model. The proteome is a highly dynamic system. The expression of proteins, their abundance, the state of modification and the subcellular localization are influenced by various parameters. Thus proteomics has become the method of choice to gain information about numerous physiological as well as pathological processes. To focus on signalling processes in oviductal epithelial cells, particularly proteins embedded in cellular signalling namely phosphorylated proteins and components of lipid raft domains (specialised microdomains of the plasma membrane that form signalling platforms) will be studied in response to the interaction with spermatozoa. Moreover, a recently established in situ biotinylation technique will be utilized to enrich cell surface proteins of epithelial cells. In a further step the extracted and purified proteins will be subjected to two-dimensional difference gel electrophoresis and vacuum matrix-assisted laser desorption/ionisation tandem mass spectrometry for protein identification. To verify the obtained data and its relevance for fertilization an in vitro co-culture system will be established. The information about proteins modulated by the physical interaction between the two different cell types will allow us to get a deeper insight into this highly relevant step. The knowledge about the signalling processes, which are necessary to generate an optimal environment for fertilization may further help us to improve in vitro techniques.

Keywords:
oviduct, epithelial cells, spermatozoa, communication, proteomics