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Mechanisms of embryo-maternal

Coordinator: Prof. Dr. Eckhard Wolf, Genzentrum der LMU-München

Unit 478

Project 2:

Proteomics-based investigations of interactions between preimplantation bovine embryos and uterine epithelial cells

Georg Arnold

During the first funding period we were focused on embryo-induced alterations in the bovine endometrium during the pre-attachment period (Day 18). Using modern holistic proteome analysis, endometrium samples derived from pregnant and non-pregnant animals as well as samples of endometrium/embryo co-culture experiments were investigated (Berendt et al.). To allow a fast and effective validation of our candidate proteins as well as the validation of candidates found by project partners, a set of antibodies was generated and characterized. These antibodies allow a fast and effective detection and quantification of candidate proteins in a higher number of samples and enable their immunhistochemical localization.
During the second funding period we will analyze endometrium samples from additional stages of gravidity using quantitative LC-MS/MS methods to get a kinetic view of alteration in the endometrium proteome during early gravidity. In a second approach we will make use of recently introduced ultrasensitive fluorophores (DIGE saturation labels) and expand our investigations to the bovine embryo proteome.

In detail our work program includes:

a) Comparative proteome analysis of four different development stages of blastocytes.

b) LC-MS based analysis of endometrium proteomes during early gravidity.

c) Investigation of Interferon tau induced alterations in the bovine endometrium using stable isotope labelling and LC-MS techniques.

d) Targeted validation of protein candidates using antibodies generated in the first funding period.

Berendt FJ, Fröhlich T, Schmidt SEM, Reichenbach HD, Wolf E, Arnold GJ (2005) Holistic differential analysis of embryo-induced alterations in the proteome of bovine endometrium in the pre-attachment period. Proteomics 5, 2551-2560

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