H M Yathish, Kumar Subodh, Chaudhary Rajni, Mishra Chinmoy, A Sivakumar, Kumar Amit, Chauhan Anuj, Ghosh S K, Mitra Abhijit
Division of Animal Genetics, ICAR-Indian Veterinary Research Institute (IVRI), Izatnagar, Bareilly, 243 122, Uttar Pradesh, India.
Division of Animal Genetics, ICAR-Indian Veterinary Research Institute (IVRI), Izatnagar, Bareilly, 243 122, Uttar Pradesh, India.
Anim Reprod Sci. 2018 Jun;193:126-139. doi: 10.1016/j.anireprosci.2018.04.060. Epub 2018 Apr 10.
Protamines (PRMs), important proteins of chromatin condensation in spermiogenesis, are promising candidate genes to explore markers of sperm motility. The coding and in-silico predicted promoter regions of these genes were investigated in 102 crossbred and 32 purebred cattle. Also, mRNA quantification was done to explore its possibility as diagnostic tool of infertility. The PCR-SSCP analysis indicated there were two band patterns only in fragment I of the PRM1 and fragment II of the PRM2 gene. The sequence analysis revealed A152G and G179A transitions in the PRM1 gene. Similarly, G35A, A49G and A64G transitions were identified in the PRM2 gene which resulted in altered amino acid sequences from arginine (R) to glutamine (Q), from arginine (R) to glycine (G) and from arginine (R) to glycine (G), respectively. This caused the reduction in molecular weight of PRM2 from 2157.66 to 1931.33 Da due to reduction in the number of basic amino acids. These altered properties of the PRM2 protein led to the reduction in Mass Motility (MM: P < 0.01), Initial Progressive Motility (IPM; P < 0.05) and Post Thaw Motility (PTM; P < 0.05) in crossbred bulls. The least squares analysis of variance indicated there was an effect of PRM2 haplotypes on MM (P = 0.0069), IPM (P = 0.0306) and PTM (P = 0.0500) in crossbred cattle and on PTM (P = 0.0408) in the overall cattle population. Based on the RT-qPCR analysis, however, there was not any significant variation of PRM1 and PRM2 gene expression among sperm of Vrindavani bulls with relatively lesser and greater sperm motility.
鱼精蛋白（PRMs）是精子发生过程中染色质浓缩的重要蛋白质，是探索精子活力标志物的有前景的候选基因。在102头杂交牛和32头纯种牛中研究了这些基因的编码区和计算机预测的启动子区域。此外，进行了mRNA定量分析，以探索其作为不孕症诊断工具的可能性。PCR-SSCP分析表明，仅在PRM1基因的片段I和PRM2基因的片段II中存在两种条带模式。序列分析揭示了PRM1基因中的A152G和G179A转换。同样，在PRM2基因中鉴定出G35A、A49G和A64G转换，这分别导致氨基酸序列从精氨酸（R）变为谷氨酰胺（Q）、从精氨酸（R）变为甘氨酸（G）以及从精氨酸（R）变为甘氨酸（G）。由于碱性氨基酸数量减少，这导致PRM2的分子量从2157.6６Da降至1931.33Da。PRM2蛋白的这些改变的特性导致杂交公牛的群体运动能力（MM：P<0.01）、初始前进运动能力（IPM；P<0.05）和解冻后运动能力（PTM；P<0.05）降低。最小二乘方差分析表明，PRM2单倍型对杂交牛的MM（P=0.0069）、IPM（P=0.0306）和PTM（P=0.0500）以及整个牛群的PTM（P=0.0408）均有影响。然而，基于RT-qPCR分析，在精子活力相对较低和较高的弗林达瓦尼公牛的精子中，PRM1和PRM2基因表达没有任何显著差异。
