PDF(Pubmed)
Abstract：
Global warming has a significant impact on the dairy farming industry, as heat stress causes reproductive endocrine imbalances and leads to substantial economic losses, particularly in tropical-subtropical regions. The Holstein breed, which is widely used for dairy production, is highly susceptible to heat stress, resulting in a dramatic reduction in milk production during hot seasons. However, previous studies have shown that cells of cows produced from reconstructed embryos containing cytoplasm (o) from Taiwan yellow cattle (Y) have improved thermotolerance despite their nuclei (n) being derived from heat-sensitive Holstein cattle (H). Using spindle transfer (ST) technology, we successfully produced ST-Yo-Hn cattle and proved that the thermotolerance of their ear fibroblasts is similar to that of Y and significantly better than that of H (p < 0.05). Despite these findings, the genes and molecules responsible for the different sensitivities of cells derived from ST-Yo-Hn and H cattle have not been extensively investigated. In the present study, ear fibroblasts from ST-Yo-Hn and H cattle were isolated, and differentially expressed protein and gene profiles were compared with or without heat stress (hs) (42 °C for 12 h). The results revealed that the relative protein expression levels of pro-apoptotic factors, including Caspase-3, -8, and -9, in the ear fibroblasts from the ST-Yo-Hn-hs group were significantly lower (p < 0.05) than those from the H-hs group. Conversely, the relative expression levels of anti-apoptotic factors, including GNA14 protein and the CRELD2 and PRKCQ genes, were significantly higher (p < 0.05) in the ear fibroblasts from the ST-Yo-Hn-hs group compared to those from the H-hs group. Analysis of oxidative phosphorylation-related factors revealed that the relative expression levels of the GPX1 gene and Complex-I, Complex-IV, CAT, and PGLS proteins were significantly higher (p < 0.05) in the ear fibroblasts from the ST-Yo-Hn-hs group compared to those from the H-hs group. Taken together, these findings suggest that ear fibroblasts from ST-Yo-Hn cattle have superior thermotolerance compared to those from H cattle due to their lower expression of pro-apoptotic factors and higher expression of oxidative phosphorylation and antioxidant factors. Moreover, this improved thermotolerance is attributed, at least partially, to the cytoplasm derived from more heat-tolerant Y cattle. Hence, using ST technology to produce more heat-tolerant H cattle containing Y cytoplasm could be a feasible approach to alleviate the negative impacts of heat stress on dairy cattle in tropical-subtropical regions.
摘要：
全球变暖对奶牛养殖业产生重大影响,由于热应激导致生殖内分泌失衡并导致巨大的经济损失，特别是在热带-亚热带地区。Holstein品种,广泛用于乳制品生产，极易受到热应力的影响，导致在炎热季节牛奶产量大幅减少。然而,先前的研究表明，尽管细胞核（n）来自对热敏感的荷斯坦牛（H），但由台湾黄牛（Y）的含有细胞质（o）的重建胚胎产生的母牛的细胞却提高了耐热性。采用主轴传递（ST）技术，我们成功生产了ST-Yo-Hn牛，并证明其耳成纤维细胞的耐热性与Y相似，并且明显优于H（p<0.05）。尽管有这些发现,尚未广泛研究负责ST-Yo-Hn和H牛的细胞的不同敏感性的基因和分子。在本研究中,分离了来自ST-Yo-Hn和H牛的耳成纤维细胞，在有或没有热应激（hs）（42°C持续12小时）的情况下，比较了差异表达的蛋白质和基因谱。结果表明，促凋亡因子的相对蛋白表达水平，包括Caspase-3，-8和-9在内，在ST-Yo-Hn-hs组的耳成纤维细胞中明显低于H-hs组（p<0.05）。相反，抗凋亡因子的相对表达水平，包括GNA14蛋白和CRELD2和PRKCQ基因，与来自H-hs组的那些相比,来自ST-Yo-Hn-hs组的耳成纤维细胞显著更高(p<0.05)。对氧化磷酸化相关因子的分析表明，GPX1基因和Complex-I的相对表达水平，Complex-IV,CAT,和PGLS蛋白在来自ST-Yo-Hn-hs组的耳成纤维细胞中显著高于(p<0.05)来自H-hs组的那些。一起来看,这些发现表明，与H牛相比，ST-Yo-Hn牛的耳成纤维细胞具有更好的耐热性，因为它们的促凋亡因子表达较低，氧化磷酸化和抗氧化因子表达较高。此外，这种改进的耐热性归因于，至少部分地,来自更耐热的Y牛的细胞质。因此，利用ST技术生产含Y细胞质的耐热H牛可能是减轻热带亚热带地区热应激对奶牛的负面影响的可行方法。
