【摘要】 Wnt-4基因是Wnt基因家族重要的成员,在许多生物事件中都发挥着重要的作用。从中华绒螯蟹(Eriocheir sinensis)卵巢转录组数据库中筛选到EsWnt-4的cDNA序列1027 bp。RT-PCR检测发现EsWnt-4在神经组织、心脏和性腺中有表达,说明在中华绒螯蟹中EsWnt-4可能与多种组织细胞的活动有关。荧光定量检测发现,受精卵中EsWnt-4 mRNA的含量较成熟卵细胞和2、4、8细胞胚胎时期高,推测EsWnt-4基因有可能在受精卵中已经开始表达,而在之后的2、4、8细胞时期其表达又受到抑制。此外,EsWnt-4在16细胞期表达量再次升高,说明该时期可能有相关的生物学事件需要EsWnt-4基因的参与。在幼体阶段EsWnt-4在第二溞状幼体及第五溞状幼体阶段表达最高,说明EsWnt-4可能与幼体体节生成及变态发育有关。更多还原

【Abstract】 Wnt-4 gene is an important member of the Wnt gene family and plays an important role in many biological events. The EsWnt-4 cDNA was obtained from the transcriptome data of Eriocheir sinensis ovary and the length of EsWnt-4 cDNA contains 1027 nucleotides,which encodes 263 deduced amino acid residues. the cDNA contains 1027 nucleotides. Multiple sequence alignment revealed that EsWnt-4 shared the highest similarity(98%) with that of the Euperipotaidei kanangrensis in conserved domains. Further,a phylogenetic tree of WNT amino acid sequences from various species was constructed. EsWnt-4 was clustered into one branch with Wnt-4 of other species. RT-PCR analysis showed that EsWnt-4 was expressed in neural organization,heart and gonadal,suggesting that EsWnt-4 was related to the activity of various of tissues.While EsWnt-4 expression was different among different tissues,it has the highest expression level in heart and brain,the lowest in thoracic ganglion. Interestingly,significantly higher expression level of EsWnt-4 was detected in fertilized eggs than in mature eggs,2-cell stage,4-cell stage,and 8-cell stage,suggesting that EsWnt-4 gene might have started expression in the spermatozoon,but the expression was suppressed in the later 2-cell,4-cell,8-cell stage. In addition,the EsWnt-4 increased in 8-16 cell stages,suggesting that EsWnt-4 might participate or play role in related biological events in this period. In the larval stage,significantly higher expression level of EsWnt-4 was detected in zoea-2 larvae and zoea-5 larvae,indicted that EsWnt-4 could be related to segment formation and metamorphosis.更多还原