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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