厚壳贻贝Wnt4基因时空表达

为探究Wnt4基因在厚壳贻贝幼虫发育阶段和组织生长过程中的作用,通过RACE技术克隆了厚壳贻贝Wnt4基因c DNA全长序列,该序列全长3342 bp,开放阅读框为1074 bp,编码357个氨基酸。该序列与人、小鼠、海胆、栉孔扇贝和长牡蛎等物种的同源性分别为61%、61%、60%、71%和76%。通过实时荧光定量PCR(q RT-PCR)分析Wnt4基因在厚壳贻贝成体多个组织中(外套膜、闭壳肌、鳃、雌雄性腺、足和消化腺)均有表达,其中在外套膜中表达量最高,推测可能与贝壳形成有关;Wnt4基因在厚壳贻贝幼虫发育阶段高表达主要集中在壳顶期,并推测Wnt4基因可能参与了贝壳形态结构发生转变的过程以及某些器官的形成与发育。本研究为进一步开展双壳贝类Wnt基因家族的功能研究提供了理论依据。

Temporal-spatial expression of Wnt4 gene in the mussel Mytilus coruscus

To understand the potential mechanisms of the Wnt4 gene at different development stages and adult tissues of the mussel Mytilus coruscus, the Wnt4 gene was cloned by RACE technique. The full length of Wnt4 gene was 3 342 bp with 1 074 bp of open reading frame (ORF) encoding 357 amino acids. Homologous analysis of the amino acids of Wnt4 gene in M. coruscus shared high similarity with Homo sapiens (61%), Mus musculus (61%), Paracentrotus lividus (61%), Chlamys farreri (71%) and Crassostrea gigas (76%). qRT-PCR showed that Wnt4 mRNA expression was detected in all tissues (mantle, adductor muscle, gill, foot, digestive gland, male and female gonad) and the highest expression was found in mantle, indicating that Wnt4 gene may play an important role in shell formation. Relative gene expression of Wnt4 during larval development stages including trochophore, D-shaped, umbo, pediveliger and the juvenile stage was also detected. The Wnt4 mRNA was highest expressed in umbo stage larvae, suggesting that Wnt4 gene may participate in the process of shell morphological structure change and certain organ formation and development. Taken together, the finding provides a new insight into investigating the function of Wnt gene family.