Yang Bingye, Li Lingling, Pu Fei, You Weiwei, Huang Heqing, Ke Caihuan
Xiamen Medical College, Xiamen, 361005, People's Republic of China,
Mol Biol Rep. 2015 May;42(5):963-75. doi: 10.1007/s11033-014-3833-y. Epub 2014 Nov 16.
Caspases have been demonstrated to possess important functions in apoptosis and immune system in vertebrate. But there is less information reported on the oyster larval development. In the present work, two full-length molluscan caspase genes, named Cacaspase-2 and Cacaspase-3, were characterized for the first time from Fujian oyster, Crassostrea angulata. Which respectively encode two predicted proteins both containing two caspase domains of p20 and p10 including the cysteine active site pentapeptide "QACRG" and the histidine active site signature. Otherwise Cacaspase-2 also contains a caspase recruitment domain. Homology and phylogenetic analysis showed that Cacaspase-2 shared high similarity with initiator caspase-2 groups, but Cacaspase-3 clustered together with executioner caspase-3 groups. Cacaspase-2 and Cacaspase-3 mRNA were both highly expressed in gills and labial palp and were significantly expressed highly in larvae during settlement and metamorphosis. Through the whole mount in situ hybridization, the location of Cacaspase-2 is in the foot of the oyster larvae and the location of Cacaspase-3 is in both the foot and velum tissues. These results implied that Cacaspase-2 and Cacaspase-3 genes play a key role in the loss of foot and Cacaspase-3 gene has an important function in the loss of velum during larvae metamorphosis in C. angulata.
半胱天冬酶已被证明在脊椎动物的细胞凋亡和免疫系统中具有重要功能。但关于牡蛎幼虫发育的报道较少。在本研究中,首次从福建牡蛎(Crassostrea angulata)中鉴定出两个全长软体动物半胱天冬酶基因,命名为Cacaspase-2和Cacaspase-3。它们分别编码两种预测的蛋白质,均包含p20和p10的两个半胱天冬酶结构域,包括半胱氨酸活性位点五肽“QACRG”和组氨酸活性位点特征序列。此外,Cacaspase-2还包含一个半胱天冬酶募集结构域。同源性和系统发育分析表明,Cacaspase-2与起始半胱天冬酶-2组具有高度相似性,但Cacaspase-3与执行半胱天冬酶-3组聚集在一起。Cacaspase-2和Cacaspase-3的mRNA在鳃和唇触须中均高度表达,并且在幼虫附着和变态期间显著高表达。通过全组织原位杂交,Cacaspase-2的定位在牡蛎幼虫的足部,Cacaspase-3的定位在足部和缘膜组织中。这些结果表明,Cacaspase-2和Cacaspase-3基因在幼虫变态过程中足部消失中起关键作用,而Cacaspase-3基因在缘膜消失中具有重要功能。