ICAR-NDRI, Karnal, Haryana, 132001, India.
ICAR-National Bureau of Animal Genetics Resources, GT Road Bypass, Karnal, Haryana, 132001, India.
Cell Stress Chaperones. 2017 Sep;22(5):675-684. doi: 10.1007/s12192-017-0770-4. Epub 2017 Mar 6.
Heat stress proteins assist cellular proteins in the acquisition of native structure. The present research was conducted to study how thermo-tolerance is modulated by HSP90 and HSP70 gene polymorphism and its association with hemato-physio-biochemical parameters, supported by their expression profiles in Chokla, Magra, Marwari, and Madras Red sheep breeds. Least square analysis revealed significant effect (P < 0.05) of season and breed on all the physiological parameters, i.e., temperature, respiratory rate, and pulse rate (a.m. and p.m.), as well as hematological parameters like Hb, packed cell volume, total erythrocyte count (TEC), neutrophil/lymphocyte (N/L) ratio, and total leukocyte count (TLC). There was a significant influence (P < 0.05) of breed on biochemical parameters such as glucose, SGOT, phosphorous, triglyceride, and cholesterol. Eight fragments were amplified and sequenced in HSP90, and 70 genes and 13 single-nucleotide polymorphisms (SNPs) were identified. Tetra-primer amplification refractory mutation system PCR, PCR-RFLP, and allele-specific PCR genotyping protocols were developed for large-scale genotyping of five SNPs. A significant difference (P < 0.05) of rectal temperature (a.m.), respiratory rate (p.m.), triglyceride, and total protein was observed at SNP01; albumin at SNP2; pulse rate (p.m.) at SNP3; and rectal temperature (p.m.), pulse rate (p.m.), Hb (g/dL), and N/L ratio at SNP4 and TLC at SNP5. Gene expression analysis revealed higher expression in less adapted animals with Madras Red < Magra < Chokla < Marwari expression pattern [corrected]. Predominant allele was found to be superior in most of the SNPs (SNP1-4) indicating the selection acting in directional manner (positive selection). Finally, it is concluded that TACCA haplotype combination of SNP1-SNP2-SNP3-SNP4-SNP5 might be of some selection advantage for the identification of animals more adaptable to heat stress.
热应激蛋白有助于细胞蛋白获得天然结构。本研究旨在研究 HSP90 和 HSP70 基因多态性如何调节耐热性及其与血液生理生化参数的关系,并通过 Chokla、Magra、Marwari 和 Madras Red 绵羊品种的表达谱来支持这一研究。最小二乘法分析表明,季节和品种对所有生理参数(如体温、呼吸率和脉搏率(上午和下午))以及血液学参数(如 Hb、红细胞压积、总红细胞计数(TEC)、中性粒细胞/淋巴细胞(N/L)比值和总白细胞计数(TLC))都有显著影响(P<0.05)。品种对生化参数(如葡萄糖、SGOT、磷、甘油三酯和胆固醇)也有显著影响(P<0.05)。在 HSP90 中扩增和测序了 8 个片段,鉴定出 70 个基因和 13 个单核苷酸多态性(SNP)。开发了四引物扩增受阻突变系统 PCR、PCR-RFLP 和等位基因特异性 PCR 基因分型方案,用于对五个 SNP 进行大规模基因分型。在 SNP01 观察到直肠温度(上午)、呼吸率(下午)、甘油三酯和总蛋白、SNP2 白蛋白、SNP3 脉搏率(下午)、SNP4 直肠温度(下午)、脉搏率(下午)、Hb(g/dL)和 N/L 比值以及 SNP5 的 TLC 有显著差异(P<0.05)。基因表达分析显示,Madras Red<Magra<Chokla<Marwari 的表达模式下,适应能力较低的动物表达水平更高。主要等位基因在大多数 SNP(SNP1-4)中表现出优势,表明选择作用是定向的(正向选择)。最后得出结论,SNP1-SNP2-SNP3-SNP4-SNP5 的 TACCA 单倍型组合可能对识别对热应激更适应的动物具有一定的选择优势。