Tong Qiaoqiong, Zhang Mengmeng, Cao Xiao, Xu Shanliang, Wang Danli, Zhao Yunlong
School of Marine Sciences, Ningbo University, Ningbo 315211, China.
School of Marine Sciences, Ningbo University, Ningbo 315211, China.
Gene. 2017 Nov 15;634:37-46. doi: 10.1016/j.gene.2017.08.035. Epub 2017 Sep 1.
Death-mediating proteases such as Caspases have been implicated in aging. Remarkably, active Caspase-3 can trigger widespread damage and degeneration, playing a key role in causing cell death. In order to explore the relationship between Caspase-3 and aging in Daphnia pulex, we cloned and analyzed the full-length cDNA sequence of its Caspase-3 gene. Both mRNA expression and activity of D. pulex Caspase-3 increased with age. Moreover, different forms of Caspase-3 appeared with aging. The expression of casp3-L was higher and decreased with age, while that of casp3-S was weak and increased with age, consistent with the trend in Caspase-3 activity. Mhc mRNA expression declined over time and was negatively correlated with age and Caspase-3. In situ hybridization results showed that Caspase-3 mRNA was expressed in different growth and reproduction stages, and its expression levels in embryos and larva were lower than that in adult D. pulex. Western blot analysis revealed the presence of Caspase-3 in the form of zymogens with a molecular weight of ~36kDa. Overall, this study explored age-associated gene regulation to provide a basis for the molecular mechanism of D. pulex reproductive conversion.
诸如半胱天冬酶等介导死亡的蛋白酶与衰老有关。值得注意的是，活性半胱天冬酶-3可引发广泛的损伤和退化，在导致细胞死亡中起关键作用。为了探究大型溞中半胱天冬酶-3与衰老之间的关系，我们克隆并分析了其半胱天冬酶-3基因的全长cDNA序列。大型溞半胱天冬酶-3的mRNA表达和活性均随年龄增长而增加。此外，随着衰老出现了不同形式的半胱天冬酶-3。casp3-L的表达较高且随年龄下降，而casp3-S的表达较弱且随年龄增加，这与半胱天冬酶-3活性的趋势一致。Mhc mRNA表达随时间下降，且与年龄和半胱天冬酶-3呈负相关。原位杂交结果表明，半胱天冬酶-3 mRNA在不同的生长和繁殖阶段均有表达，其在胚胎和幼虫中的表达水平低于成年大型溞。蛋白质免疫印迹分析显示存在分子量约为36kDa的酶原形式的半胱天冬酶-3。总体而言，本研究探索了与年龄相关的基因调控，为大型溞生殖转化的分子机制提供了依据。
