Identification of ADAM10 and ADAM17 with potential roles in the spermatogenesis of the Chinese mitten crab, Eriocheir sinensis
Identification of ADAM10 and ADAM17 with potential roles in the spermatogenesis of the Chinese mitten crab, Eriocheir sinensis
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鉴定 ADAM10 和 ADAM17 在中华绒螯蟹精子发生中的潜在作用
DOI：
10.1016/j.gene.2015.02.060
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发表时间：
2015-05-10
期刊：
影响因子：
3.5
通讯作者：
Wang, Qun
中科院分区：
文献类型：
--
作者：
Li, Qing;Xie, Jing;Wang, Qun
ADAM（一种崩解素和金属蛋白酶）家族在精子和卵子的融合、发育、炎症、粘附和迁移中起重要作用。ADAM10和ADAM17参与精子发生。为了更好地了解ADAM10和ADAM17在中华绒螯蟹（Eriocheir sinensis）中的作用，我们克隆了ADAM10和ADAM17的全长cdna，分别命名为Es-ADAM10和Es-ADAM17。序列和结构分析表明，Es-ADAM10和Es-ADAM17具有ADAM家族的典型结构。实时定量逆转录聚合酶链反应分析显示，Es-ADAM10和Es-ADAM17 mrna分布在心脏、肝胰脏、肠、脑、肌肉、胸神经节、血淋巴、胃、睾丸、卵巢、鳃和副腺。这两种mrna在肌肉中均高表达，在睾丸、卵巢和副腺中表达相对较高。Es-ADAM17 mRNA在睾丸发育各阶段均有表达，7 - 9月较高，10 - 11月最低，12 - 1月逐渐升高，1月达到峰值。相比之下，Es-ADAM10 mRNA的表达在睾丸发育过程中是恒定的。免疫荧光进一步显示，Es-ADAM10和Es-ADAM17蛋白存在于精母细胞的细胞质和细胞膜中，在精子中均检测到。此外，依托波苷诱导Es-ADAM17和Es-ADAM10 mRNA和蛋白水平上调。本研究首次发现Es-ADAM10和Es-ADAM17也参与了中华绒螯蟹的精子发生，主要参与了后期生殖细胞的凋亡。(C) 2015 Elsevier B.V.版权所有
The ADAM (a disintegrin and metalloprotease) family plays an important role in sperm and egg fusion, development, inflammation, adhesion and migration. ADAM10 and ADAM17 are involved in the spermatogenesis. To better understand the role of ADAM10 and ADAM17 in the Chinese mitten crab, Eriocheir sinensis, the full-length cDNAs of ADAM10 and ADAM17 were cloned, and named Es-ADAM10 and Es-ADAM17, respectively. Sequence and structural analysis showed that Es-ADAM10 and Es-ADAM17 have the typical structure of the ADAM family. Quantitative real-time reverse transcription polymerase chain reaction analysis showed that Es-ADAM10 and Es-ADAM17 mRNAs were distributed in the heart, hepatopancreas, intestines, brain, muscle, thoracic ganglia, hemolymph, stomach, testis, ovary, gill and accessory gland. Both mRNAs were highly expressed in the muscles, and relatively high in the testis, ovary and accessory gland. In addition, the Es-ADAM17 mRNA level was detected in every stage of testis development, being relatively high from July to September, the lowest during October and November, increasing from December to January, and reached a peak in January. By contrast, the expression of Es-ADAM10 mRNA was constant during testis development. Immunofluorescence further showed that Es-ADAM10 and Es-ADAM17 proteins were present in the cytoplasm and cytomembrane of spermatocytes, and both detected in the sperm. Furthermore, etoposide induced upregulation of Es-ADAM17 and Es-ADAM10 at both the mRNA and protein levels. This study first showed that Es-ADAM10 and Es-ADAM17 were also involved in the spermatogenesis and mainly participated in the later germ cell apoptosis in E. sinensis. (C) 2015 Elsevier B.V. All rights reserved.