半滑舌鳎Dorsalin-1-like基因的克隆与表达

摘要:  Dorsalin-1 是骨形态发生蛋白(bone morphogenetic proteins, BMPs)家族成员,  在调控脊椎动物神经系统的发
育中具有重要作用。目前,  国内外有关脊椎动物 Dorsalin-1 的研究报道十分有限。本研究克隆了半滑舌鳎
(Cynoglossus semilaevis) Dorsalin-1-like基因的cDNA序列,  并分析了Dorsalin-1-like mRNA在半滑舌鳎成鱼的组织
表达差异及其在早期发育阶段的表达水平变化。 研究结果表明,  半滑舌鳎Dorsalin-1-like基因cDNA全长为1 961 bp,
包括了 153 bp 的 5′非翻译区(5′UTR)、 编码 419 个氨基酸的 1 260 bp 的开放读码框(ORF),  以及 548 bp  的 3′非翻译
区(3′UTR)。 同源性分析和分子进化聚类分析结果显示,  半滑舌鳎 Dorsalin-1-like 鲀 与红鳍东方 (Taleifugu rubripes)、
尼罗罗非鱼(Oreochromis niloticus)等的同源性最高,  并与之共同聚为鱼类 Dorsalin-1 分支。此外, Dorsalin-1-like
mRNA 在半滑舌鳎成鱼的 13 个组织中具有广泛的分布,  并在心脏组织中表达量最高,  在肝、脊髓等组织中次之。
在半滑舌鳎早期发育阶段(卵期、仔鱼、稚鱼、幼鱼)中, Dorsalin-1-like 在卵期中的胚胎期表达量最高,  其次为前期
仔鱼期。上述实验结果表明, Dorsalin-1 在调控卵期和仔鱼期的早期发育中具有重要作用,  并可能通过调节神经系
统的发育进而参与心脏、骨骼等组织的发育调控。本研究旨在揭示 Dorsalin-1-like 基因在调控半滑舌鳎早期神经
系统发育的重要作用,  并为筛选半滑舌鳎骨骼发育的调控因子奠定理论基础。

Molecular cloning and expression analysis of Dorsalin-1-like gene in the tongue sole (Cynoglossus semilaevis)

Abstract: As a member of the bone morphogenetic protein (BMP) family, Dorsalin-1 plays an important role in
regulating the development of the nervous system. To date, studies involving the vetebrate Dorsalin-1 has thus far
only rarely been reported. In this study, molecular cloning and expression analysis of Dorsalin-1 gene were
performed in the tongue sole (Cynoglossus semilaevis). As a result, the Dorsalin-1-like cDNA sequence is 1 961
bp, which contains a 5’-untranslated region (UTR) of 153 bp, an open reading frame (ORF) of 1 260 bp encoding a
protein of 419 amino acids, and a 3’-untranslated region (UTR) of 548 bp. The homology and phylogenetic
analysis revealed that the deduced amino acid sequence of C. semilaevis Dorsalin-1-like gene exhibited the highest
identity with those of Takifugu rubripes and Oreochromis niloticus, and the fish Dorsalin-1 formed a single lineage
from the other vertebrates. The tissue expression distribution pattern of Dorsalin-1-like gene was examined in the
13 tissues of female and male adults. The results showed that the Dorsalin-1-like mRNA was predominantly
expressed in heart, and less expressed in the liver and spinal cord. Finally, the Dorsalin-1-like mRNA levels at
early developmental stages were also detected at egg, larva, juvenile and fingerling stages. The highest level was
observed in the embryonic stage (egg stage) and early-stage larvae (larva stage). These findings suggest that
Dorsalin-1 may play an important role in regulating early development of the egg and larva stages, and maybe
involved in the heart and skeletal development through the regulation of the nervous system.