Differential proteomic profiling of X and Y sperm enriched Sahiwal bull semen
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Date
2020-10
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G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand)
Abstract
Pre-determining the foetal sex is of prime importance for economic and scientific growth of both dairy farmers and livestock owner. Hence, the present study was aimed to compare the protein profile of X- and Y-sorted semen of Sahiwal bull. Semen sample (n=6) were categorized into three groups i.e. group I (X-sorted), group II (Y-sorted) and control group (both X- and Y- sperms). Total protein estimation SDS PAGE and nano LC-MS of all three samples was performed. In this study, significant total protein concentration in control group, X-sorted and Y sorted Sahiwal semen was 2.189 ± 0.035 mg/ml, 2.098 ± 0.033 mg/ml, and 1.79 ± 0.042 mg/ml respectively. Significant difference (P < 0.05) was observed in total protein concentration between control group and Y-sorted semen. Also, between X-sorted and Y sorted semen the difference was found to be significant (P < 0.05) at 5% level of significance. In SDS PAGE, specific proteins of molecular weight between 18-24KDa and between 30-37KDa were present in X-sorted sperms. Also, band corresponding to 25KDa was specific to Y-sorted sperms. In Nano LC/MS, total 241 proteins were identified, out of which 113 were differentially expressed between X- and Y-sorted sperms, in which 54 proteins are having at
least two unique peptides. Out of 54 proteins, 27 were upregulated in X-sorted sample, 3 were upregulated in Y-sorted sample and 24 were differentially downregulated. Highly upregulated protein viz. Armadillo repeat containing 12 protein, NDC1 transmembrane nucleoporin, Beta-nerve growth factor, C-type natriuretic peptide, Nucleobindin-2, Phosphoglycerate mutase 2, Calmodulin along with one uncharacterised protein having accession number F1MN9 may have potential to be used as biomarker for separating X and Y sperm.