引用本文:	刘万,张晓臣,赵春暖,张浩瑜,杨李俊淞,韩怡静,高雁,王晓通.microRNA-183-5p通过调控MITF基因影响长牡蛎黑色素形成[J].海洋科学,2025,49(7):63-72.

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*microRNA-183-5p通过调控MITF基因影响长牡蛎黑色素形成*
刘万¹, 张晓臣¹, 赵春暖², 张浩瑜¹, 杨李俊淞¹, 韩怡静¹, 高雁², 王晓通¹
1.鲁东大学水产学院, 山东烟台 264025;2.烟台市海洋经济研究院, 山东烟台 264003

摘要:

黑色素的合成是一个极为复杂的过程, 多种基因及microRNAs(miRNAs)都参与其中。近年来, 越来越多的研究证实miRNAs在哺乳动物毛色生成过程中发挥着重要作用, 但miRNA对于贝类参与黑色素合成的调控机制研究较少。其中, 长牡蛎(Crassostrea gigas)外套膜与黑色素沉着密切相关, 是壳色形成及黑色素产生的关键部位。本研究采用RNAhybrid预测miR-183-5p的靶基因; 通过实时荧光定量PCR(quantitative real time polymerase chain reaction, qRT-PCR)、双荧光素酶报告基因检测、和马松丰塔纳黑色素染色技术等验证miR-183-5p对黑色素合成的影响。软件预测和双荧光素酶报告基因检测证实小眼畸形相关转录因子(Microphthalmia-associated transcription factor, MITF)是miR-183-5p的靶基因, qRT-PCR表明miR-183-5p在长牡蛎黑色和白色外套膜中均有表达且在白色中显著表达, 其miR-183-5p过表达后使黑色素形成相关基因表达量显著下调, 染色结果也显示模拟物组棕色颗粒较对照组明显减少。综上, miR-183-5p可能通过调控MITF的表达影响长牡蛎黑色素的形成。

关键词: 长牡蛎(Crassostrea gigas) 黑色素 miR-183-5p MITF

DOI：10.11759/hykx20250225001

分类号:S917.4

基金项目:国家自然科学基金资助项目(42076088);山东省自然科学基金资助项目(ZR2023QC107,ZR2022QC233);山东省重点研发计划资助项目(2022LZGC015);山东省泰山学者专项资金项目(tstp20240518);山东省现代农业产业技术体系项目(SDAIT-14-01);烟台市省级以上领军人才专项支持资金项目(202203);烟台市水产种业提升项目(2023HD)

Regulation of melanin formation in Crassostrea gigas by microRNA-183-5p via MITF gene

LIU Wan¹, ZHANG Xiaochen¹, ZHAO Chunnuan², ZHANG Haoyu¹, YANG Lijunsong¹, HAN Yijing¹, GAO Yan², WANG Xiaotong¹

1.School of Fisheries, Ludong University, Yantai 264025, China;2.Yaitai Marine Economic Research Institute, Yantai 264003, China

Abstract:

Melanin synthesis is an extremely complex process involving multiple genes and microRNAs (miRNAs). In recent years, numerous studies have confirmed that miRNAs play important roles in mammalian coat color formation. However, only a few studies have examined the mechanisms by which miRNAs regulate melanin synthesis in shellfish. In Crassostrea gigas (C. gigas), the mantle is closely related to melaninosis, a key component of shell color formation and melanin production. In this study, RNAhybrid was used to predict the target genes of miR-183-5p. The effects of miR-183-5p on melanin synthesis were verified by quantitative real-time polymerase chain reaction (qRT-PCR), dual-luciferase reporter gene assays, and Masson-Fontana melanin staining. In silico predictions and dual-luciferase reporter gene assays confirmed that microphthalmia-associated transcription factor (MITF) is a target of miR-183-5p. qRT-PCR analysis revealed that miR-183-5p is expressed at higher levels in white than in black mantles of C. gigas. The overexpression of miR-183-5p significantly downregulated genes related to melanin formation. Melanin staining showed that the brown particles in the mimic group were markedly reduced compared to those in the negative control group. In conclusion, miR-183-5p may regulate melanin formation in C. gigas by targeting MITF.

Key words: Crassostrea gigas melanin miR-183-5p MITF