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X 性控和 Y 性控萨希瓦尔公牛精液的差异蛋白质组学图谱。

Differential proteomic profile of X- and Y- sorted Sahiwal bull semen.

机构信息

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.

出版信息

Res Vet Sci. 2022 May;144:181-189. doi: 10.1016/j.rvsc.2021.11.013. Epub 2021 Nov 19.

Abstract

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.

摘要

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。

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