School of Life Science, East China Normal University, No 3663 North Zhong-Shan Road, Shanghai, China.
Fish Shellfish Immunol. 2011 Jan;30(1):263-72. doi: 10.1016/j.fsi.2010.10.016. Epub 2010 Nov 2.
Apoptosis is a central regulatory feature of the immune system, and the most common form of death among immunological cells. However, the function of apoptosis, within the innate immune system of invertebrates, remains largely unknown. For this reason, we investigated the immune functionality of two apoptosis genes, caspase and nm23, in the Chinese mitten crab (Eriocheir sinensis), which is a commercially important and disease vulnerable aquaculture species. The entire length caspase and nm23 cDNA genes were cloned using PCR, based on an initial expressed sequence tag (EST) isolated from a hepatopancreatic cDNA library. The caspase cDNA contained an 1119 bp open reading frame that encoded a putative 372 amino acid protein, while nm23 cDNA contained a 456 bp open reading frame that encoded a putative 151 amino acid protein. Comparison, with other reported invertebrate and vertebrate sequences, revealed the presence of conserved enzyme active sites that were common among caspase and nm23 superfamilies. In brief, caspase and nm23 mRNA expression in E. sinensis were (a) both detected in all tissues, including the hemocytes, heart, hepatopancreas, gill, stomach, muscle, intestine, brain and eyestalk, and (b) responsive in hemocytes, gill and hepatopancreas to a Vibrio anguillarum immuno-challenge all appeared sharp increase. Collectively, the data presented here demonstrate the successful isolation of caspase and nm23 apoptosis genes from the Chinese mitten crab, and their role in the innate immune system of an invertebrate.
细胞凋亡是免疫系统的核心调控特征，也是免疫细胞最常见的死亡形式。然而，无脊椎动物先天免疫系统中细胞凋亡的功能在很大程度上仍然未知。基于从中华绒螯蟹肝胰腺 cDNA 文库中分离出的一个初始表达序列标签 (EST)，我们使用 PCR 克隆了两个凋亡基因 caspase 和 nm23 的全长 cDNA。caspase cDNA 包含一个 1119 bp 的开放阅读框，编码一个假定的 372 个氨基酸的蛋白质，而 nm23 cDNA 包含一个 456 bp 的开放阅读框，编码一个假定的 151 个氨基酸的蛋白质。与其他报道的无脊椎动物和脊椎动物序列的比较表明，存在保守的酶活性位点，这些位点在 caspase 和 nm23 超家族中是共同的。简而言之，E. sinensis 中的 caspase 和 nm23 mRNA 表达(a)均在所有组织中检测到，包括血细胞、心脏、肝胰腺、鳃、胃、肌肉、肠、脑和眼柄，(b)在血细胞、鳃和肝胰腺中对鳗弧菌免疫挑战的反应均明显增加。总的来说，这里呈现的数据表明成功地从中华绒螯蟹中分离出了 caspase 和 nm23 凋亡基因，并证明了它们在无脊椎动物先天免疫系统中的作用。
