264. Proteomic approaches to the study of conceptus fluids from first trimester bovine pregnancies
G. A. Riding A D , S. A. Lehnert A D , A. J. French B D and J. R. Hill C DA CSIRO Livestock Industries, Queensland Bioscience Precinct, St Lucia, QLD, Australia
B Centre for Reproduction and Development, Monash Institute of Medical Research, Monsah University, Clayton, VIC, Australia
C CSIRO Livestock Industries, FD McMaster Laboratory, Armidale, NSW, Australia
D CRC for Innovative Dairy Products, Melbourne, VIC, Australia
Reproduction, Fertility and Development 17(9) 108-108 https://doi.org/10.1071/SRB05Abs264
Submitted: 26 July 2005 Accepted: 26 July 2005 Published: 5 September 2005
Abstract
Advanced reproductive technologies using in vitro production systems in cattle and other livestock species offer significant opportunities for improving the reproductive performance of the female. Impeding the development is the overall inefficiency and significant costs surrounding the high incidence of embryo wastage, particularly during early pregnancy and the occurrence of foetal developmental abnormalities, placentation irregularities and late term losses.
Effective molecular interchange and communication between the conceptus and its mother is of utmost importance for the development, growth and continued viability of the foetus. We are using a proteomics approach to determine the protein profile (proteome) of the conceptus fluids (amniotic and allantoic) during the first trimester of normal, IVF and NT pregnancies. Proteomic analysis and comparison of the proteins present in these fluids may lead to the discovery of novel biomarkers which can be used to determine foetal viability status. Collections of foetal fluids from abattoir collections of Day 35 to Day 100 pregnancies were characterised with respect to volume and total protein estimates and subjected to various fractionation procedures. Protein fingerprints of conceptus fluid fractions have been generated by two-dimensional electrophoresis and the identity of proteins determined by matrix-assisted laser desorption ionisation time-of-flight mass spectrometry and database searches. Enrichment of the low-abundance and low molecular weight proteins has been achieved by ultrafiltration under denaturing and reducing conditions.
We were able to establish the identity of the majority of high-abundance proteins and a significant proportion of low-abundance proteins in both amniotic and allantoic fluids using this method. Searching of MSDB or NCBInr mammalian protein sequence databases coupled with the putative protein database deduced from the first draft assembly of the bovine genome sequence, was achieved using Mascot software.
