Identification of potential protein biomarkers for early detection of pregnancy in cow urine using 2D DIGE and label free quantitation

Preeti Rawat, Shveta Bathla, Rubina Baithalu, Munna Lal Yadav, Sudarshan Kumar, Syed Azamal Ali, Anurag Tiwari, Masoud Lotfan, Jasmine Naru, Manoj Jena, Pradip Behere, Ashok K. Balhara, Rajesh Vashisth, Inderjeet Singh, Ajay Dang, Jai K. Kaushik, Tushar K. Mohanty, Ashok K. Mohanty

An early, reliable and noninvasive method of early pregnancy diagnosis is prerequisite for efficient reproductive management in dairy industry. The early detection of pregnancy also help in to reduce the calving interval and rebreeding time which is beneficial for industries as well as farmers. The aim of this work is to identify potential biomarker for pregnancy detection at earlier stages (16-25 days). To achieve this goal we performed DIGE and LFQ for identification of protein which has significant differential expression during pregnancy. DIGE experiment revealed a total of eleven differentially expressed proteins out of which nine were up regulated having fold change ≥1.5 in all time points. The LFQ data analysis revealed 195 differentially expressed proteins (DEPs) out of 28 proteins were up-regulated and 40 down regulated having significant fold change ≥1.5 and ≤0.6 respectively. Bioinformatics analysis of DEPs showed that a majority of proteins were involved in regulation of leukocyte immunity, endopeptidase inhibitor activity, regulation of peptidase activity and polysaccharide binding. This is first report on differentially expressed protein during various time points of pregnancy in cow to our best knowledge. In our work, we identified few proteins such MBP, SERPIN, IGF which were differentially expressed and actively involved in various activities related to pregnancy such as embryo implantation, establishment and maintenance of pregnancy. Due to their involvement in these events, these can be considered as biomarker for pregnancy but further validation of is required.