| 摘要: |
| 视黄酸X受体(Retinoid X receptor, RXR)是核受体超家族的重要成员,在后生动物变态发育过程中发挥关键调控作用,但其在长牡蛎(Crassostrea gigas)附着变态阶段是否行使功能尚不清楚。以长牡蛎眼点幼虫为研究对象,克隆获得cgRXR基因并构建GST-cgRXR融合蛋白,采用GST-pull down联合LC-MS/MS质谱分析的方法筛选并鉴定cgRXR的潜在互作蛋白,并通过Western blot对实验体系的特异性和可靠性进行验证。结果共鉴定获得57个cgRXR候选互作蛋白。GO功能注释与分类分析显示,这些蛋白在生物过程层面主要富集于翻译调控和通过剪接体介导的mRNA剪接等过程,在分子功能层面以RNA结合和ATP结合活性为主,在细胞组分层面主要定位于细胞质、细胞核以及剪接体和核糖核蛋白复合物。为进一步验证cgRXR在长牡蛎幼虫附着变态中的作用,采用抑制剂UVI3003和激动剂Bexarotene进行药理干预。结果显示,无论激活还是抑制cgRXR信号,均不同程度降低幼虫24 h和36 h附着率及36 h变态率,并呈一定的浓度依赖趋势。qRT-PCR结果表明,部分变态相关基因的表达在药物处理后发生变化。综合结果提示cgRXR参与长牡蛎幼虫附着与变态过程的调控,为进一步解析无脊椎动物核受体介导的变态调控机制提供了新的线索。 |
| 关键词: 长牡蛎 附着变态 视黄酸X受体 蛋白互作 |
| DOI：10.11693/hyhz20260300057 |
| 分类号: |
| 基金项目:国家重点研发计划项目,2022YFD2401204号；崂山实验室科技创新项目, LSKJ202203001号; 泰山学者项目, tsqn202211250号 |
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| Preliminary Study on the Mechanism of Retinoid X Receptor Involvement in Larval Settlement and Metamorphosis of the Pacific Oyster |
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Yang Mingyu, Xie Yusu, Liu Zhitong, Liu Xueqian, Xu Fei
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The Institute of Oceanology, Chinese Academy of Sciences
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| Abstract: |
| The Retinoid X Receptor (RXR) is an important member of the nuclear receptor superfamily and plays a key regulatory role in the metamorphic development of metazoans. However, whether it functions during the settlement and metamorphosis stage in the Pacific oyster remains unclear. Using eyed larvae of the Pacific oyster (Crassostrea gigas) as the research subject, the cgRXR gene was cloned, and a GST-cgRXR fusion protein was constructed. Potential interacting proteins of cgRXR were screened and identified using GST-pull down combined with LC-MS/MS mass spectrometry analysis, and the specificity and reliability of the experimental system were verified by Western blot. The results identified a total of 57 candidate interacting proteins for cgRXR. GO functional annotation and classification analysis showed that these proteins are mainly enriched in biological processes such as translational regulation and mRNA splicing via the spliceosome; in molecular functions, they are primarily associated with RNA binding and ATP binding activities; and in cellular components, they are mainly localized in the cytoplasm, nucleus, spliceosome, and ribonucleoprotein complexes. To further verify the role of cgRXR in larval settlement and metamorphosis of the Pacific oyster, pharmacological interventions were performed using the inhibitor UVI3003 and the agonist Bexarotene. The results showed that both activation and inhibition of cgRXR signaling reduced larval settlement rates at 24 h and 36 h, as well as the metamorphosis rate at 36 h, to varying degrees, exhibiting a certain concentration-dependent trend. qRT-PCR results indicated that the expression of some metamorphosis-related genes changed after drug treatment. Collectively, these findings suggest that cgRXR participates in the regulation of larval settlement and metamorphosis in the Pacific oyster, providing new clues for further elucidating the mechanisms of nuclear receptor-mediated metamorphosis regulation in invertebrates. |
| Key words: Pacific oyster settlement and metamorphosis Retinoid X Receptor protein interaction |