College of Animal Science, South China Agricultural University, Guangzhou, 510642, People's Republic of China.
Mol Biol Rep. 2013 Sep;40(9):5295-306. doi: 10.1007/s11033-013-2630-3. Epub 2013 Jun 21.
Thermo stress induces heat shock proteins (HSPs) expression and HSP90 family is one of them that has been reported to involve in cellular protection against heat stress. But whether there is any association of genetic variation in the Hsp90β gene in chicken with thermo tolerance is still unknown. Direct sequencing was used to detect possible SNPs in Hsp90β gene 5' flanking region in 3 chicken breeds (n = 663). Six mutations, among which 2 SNPs were chosen and genotypes were analyzed with PCR-RFLP method, were found in Hsp90β gene in these 3 chicken breeds. Association analysis indicated that SNP of C.-141G>A in the 5' flanking region of the Hsp90β gene in chicken had some effect on thermo tolerance traits, which may be a potential molecular marker of thermo tolerance, and the genotype GG was the thermo tolerance genotype. Hsp90β gene mRNA expression in different tissues detected by quantitative real-time PCR assay were demonstrated to be tissue dependent, implying that different tissues have distinct sensibilities to thermo stress. Besides, it was shown time specific and varieties differences. The expression of Hsp90β mRNA in Lingshan chickens in some tissues including heart, liver, brain and spleen were significantly higher or lower than that of White Recessive Rock (WRR). In this study, we presume that these mutations could be used in marker assisted selection for anti-heat stress chickens in our breeding program, and WRR were vulnerable to tropical thermo stress whereas Lingshan chickens were well adapted.
热应激诱导热休克蛋白(HSPs)的表达,其中 HSP90 家族已被报道参与细胞对热应激的保护。但是,鸡的 Hsp90β 基因的遗传变异是否与耐热性有关仍然未知。本研究采用直接测序的方法在 3 个鸡种(n=663)中检测 Hsp90β 基因 5'侧翼区可能存在的 SNP。在这 3 个鸡种的 Hsp90β 基因中发现了 6 个突变,其中选择了 2 个 SNP,并通过 PCR-RFLP 方法分析了基因型。关联分析表明,鸡 Hsp90β 基因 5'侧翼区的 SNP C.-141G>A 对耐热性性状有一定影响,可能是耐热性的潜在分子标记,基因型 GG 是耐热性基因型。通过定量实时 PCR 检测不同组织中的 Hsp90β 基因 mRNA 表达表明,其表达具有组织依赖性,这意味着不同组织对热应激有不同的敏感性。此外,还表现出时间特异性和品种差异。在心脏、肝脏、大脑和脾脏等组织中,灵山鸡的 Hsp90β mRNA 表达明显高于或低于隐性白洛克鸡(WRR)。本研究推测,这些突变可用于我们的育种计划中抗热应激鸡的标记辅助选择,而 WRR 对热带热应激敏感,而灵山鸡则适应良好。