Department of Veterinary Gynaecology and Obstetrics, College of Veterinary and Animal Sciences, G B Pant University of Agri. & Tech., Pantnagar 263145, Uttarakhand, India.
Department of Biochemistry, College of Basic Sciences and Humanities, G B Pant University of Agri. & Tech., Pantnagar 263145, Uttarakhand, India.
Res Vet Sci. 2022 May;144:181-189. doi: 10.1016/j.rvsc.2021.11.013. Epub 2021 Nov 19.
The identification of differential proteins between X- and Y-sperm may be useful for immunological sexing of sperm. Hence, the present study was aimed to compare the protein profile of X- and Y-sorted Sahiwal bull semen using SDS-PAGE and Liquid Chromatography coupled with Mass Spectrometry (Nano LC-MS). 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). SDS PAGE revealed specific proteins of molecular weight between 18 and 24 kDa and between 30 and 37 kDa were present in X-sorted sperms. Also, band corresponding to 25 kDa was specific to Y-sorted sperms. Data obtained from Nano LC/MS is analysed by search engine database i.e., MASCOT and SEQUEST HT. Total, 241 proteins were identified, out of which 113 were differentially expressed between X- and Y-sorted sperms, in which 54 proteins showed 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 in X-sperm viz. Armadillo repeat containing 12 protein, NDC1 transmembrane nucleoporin, β-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.
X 和 Y 精子之间差异蛋白的鉴定可能有助于精子的免疫性别鉴定。因此，本研究旨在使用 SDS-PAGE 和液相色谱-质谱联用（Nano LC-MS）比较 X 和 Y 分选的萨希瓦尔公牛精液的蛋白质图谱。将精液样本（n=6）分为三组，即 I 组（X 分选）、II 组（Y 分选）和对照组（X 和 Y 精子）。SDS PAGE 显示 X 分选精子中存在分子量在 18 和 24 kDa 之间和 30 和 37 kDa 之间的特定蛋白质。此外，对应于 25 kDa 的条带是 Y 分选精子特有的。从 Nano LC/MS 获得的数据通过搜索引擎数据库（MASCOT 和 SEQUEST HT）进行分析。总共鉴定出 241 种蛋白质，其中 113 种在 X 和 Y 分选精子之间表达差异，其中 54 种蛋白质显示至少有两个独特的肽。在 54 种蛋白质中，27 种在 X 分选样本中上调，3 种在 Y 分选样本中上调，24 种下调。X 精子中高度上调的蛋白包括 Armadillo repeat containing 12 protein、NDC1 跨膜核孔蛋白、β-神经生长因子、C 型利钠肽、核结合蛋白-2、磷酸甘油酸变位酶 2、钙调蛋白以及一个具有 accession number F1MN9 的未鉴定蛋白，可能具有作为分离 X 和 Y 精子的生物标志物的潜力。
