唇䱻硬骨蛋白基因克隆及其在肌间刺发生过程中的表达

MOLECULAR CHARACTERIZATION OF A SCLEROSTIN HOMOLOGUE IN BARBEL STEED (HEMIBARBUS LABEO) AND ITS RELATIONSHIP WITH INTERMUSCULAR BONE DEVELOPMENT

  • 摘要: 为探究硬化蛋白(Sclerostin, SOST)与肌间刺发生之间的关系, 研究通过转录组测序和RT-PCR获得了唇䱻(Hemibarbus labeo)硬化蛋白基因cDNA序列。序列分析表明, 硬化蛋白由信号肽和成熟肽两部分组成, 成熟肽包含一个胱氨酸结样结构域。系统进化树分析表明, 唇䱻硬化蛋白与金鱼(Carassius auratus)硬化蛋白最为接近。通过荧光定量PCR检测发现唇䱻硬化蛋白基因mRNA在所有被检测的组织中均有表达, 在鳃中的表达量最高。RNA原位杂交结果表明, 硬化蛋白mRNA分布于肌膈, 且随着肌间刺的发生信号逐步减弱。RT-qPCR显示, 在肌间刺4个发育阶段中, 硬化蛋白基因mRNA的变化显著, 在阶段Ⅰ表达量最高, 随后逐步降低。综上, 唇䱻硬化蛋白与肌间刺骨化存在一定的相关性。研究结果将为进一步调查鱼类硬化蛋白功能及硬化蛋白在肌间刺形成过程中的作用提供理论依据。

     

    Abstract: Previous studies have demonstrated that sclerostin (SOST) binds to and antagonizes the activity of bone morphogenetic proteins (BMPs). Because of the potential role of BMPs in intermuscular bone (IB) development, we explored the relationship between SOST and IB development. In this study, we identified a cDNA sequence encoding a SOST homologue in barbel steed (Hemibarbus labeo). Sequence analysis revealed that the SOST protein is composed of a signal peptide and a mature peptide which contained a C-terminal cystine knot-like domain. Phylogenetic tree analysis indicated that barbel steed SOST was closely related to goldfish (Carassius auratus) SOST. Expression of SOST transcripts were detected in all the tested tissues, with the highest transcript levels being found in the gill. in situ RNA hybridization results showed that SOST mRNA was obviously distributed in the myosepta. RT-qPCR showed that the transcript levels of the SOST gene significantly changed during the four developmental stages of IBs. This timing overlapped with IB formation, providing support for a role of SOST in IB development.

     

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