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牦牛组织和肌肉卫星细胞中 轴的表达模式与功能的综合分析。

Comprehensive analysis of the expression patterns and function of the axis in yak tissues and muscle satellite cells.

作者信息

Ma Zongliang, Chai Zhixin, Yang Huan, Zhang Xiangfei, Zhao Hongwen, Luo Xiaolin, Zhong Jincheng, Wu Zhijuan

机构信息

Qinghai-Tibetan Plateau Grass-Feeding Livestock Engineering Technology Research Center of Sichuan Province, Southwest Minzu University, Chengdu, China.

Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization Key Laboratory of Sichuan Province, Southwest Minzu University, Chengdu, China.

出版信息

Front Vet Sci. 2024 Sep 5;11:1448587. doi: 10.3389/fvets.2024.1448587. eCollection 2024.

Abstract

BACKGROUND

The long interspersed nuclear element 1 () retrotransposon has been identified as a specific substrate for fat mass and obesity-related gene (), which facilitates the removal of N-methyladenosine modifications from its targeted RNAs.

METHODS

This study examined the dynamic interaction between and in yak tissues and muscle satellite cells, utilizing RT-qPCR, RNA immunoprecipitation (RIP), immunofluorescence staining, and techniques involving overexpression and interference of and to elucidate the relationship between and in yak tissues and muscle satellite cells.

RESULTS

Cloning and analysis of the coding sequence in Jiulong yak revealed a conserved protein structure across various breeds, with notable homology observed with domestic yak, domestic cattle, and Java bison. Comprehensive examination of and gene expression patterns in Jiulong yaks revealed consistent trends across tissues in both sexes. mRNA levels were markedly elevated in the heart and kidney, while RNA was predominantly expressed in the heart. Immunoprecipitation confirmed the direct interaction between the FTO protein and RNA in yak tissues and muscle satellite cells. The - axis was confirmed by a significant decrease in RNA enrichment following its expression interference in yak muscle satellite cells. Overexpression of substantially reduced the expression of recombinant myogenic factor 5 (). However, interference had no discernible effect on and myoblast determination protein 1 () mRNA levels. Immunofluorescence analysis revealed no alterations in Ki-67 protein expression following interference or overexpression. However, phalloidin staining demonstrated enhancement in the myotube fusion rate of yak muscle satellite cells upon interference.

CONCLUSION

This comprehensive mapping of the and mRNA expression patterns establishes a direct interaction between the protein and RNA in yak. The findings suggest that overexpression promotes muscle satellite cells differentiation, whereas negatively regulates myotube fusion. The study provides fundamental insights into the role of the - axis in determining the fate of muscle satellite cells in yak, laying a solid theoretical foundation for future investigations.

摘要

背景

长散在核元件1(LINE-1)反转录转座子已被确定为脂肪量和肥胖相关基因(FTO)的特定底物,FTO可促进从其靶向RNA上去除N-甲基腺苷修饰。

方法

本研究利用RT-qPCR、RNA免疫沉淀(RIP)、免疫荧光染色以及LINE-1和FTO过表达与干扰技术,检测了牦牛组织和肌肉卫星细胞中LINE-1与FTO之间的动态相互作用,以阐明牦牛组织和肌肉卫星细胞中LINE-1与FTO的关系。

结果

九龙牦牛LINE-1编码序列的克隆与分析显示,不同LINE-1品种的蛋白质结构保守,与家牦牛、家牛和爪哇野牛具有显著同源性。对九龙牦牛中LINE-1和FTO基因表达模式的综合检测发现,两性各组织中的趋势一致。LINE-1 mRNA水平在心脏和肾脏中显著升高,而FTO RNA主要在心脏中表达。免疫沉淀证实了牦牛组织和肌肉卫星细胞中FTO蛋白与LINE-1 RNA之间的直接相互作用。在牦牛肌肉卫星细胞中干扰LINE-1表达后,LINE-1 RNA富集显著降低,证实了二者的负相关关系。FTO过表达显著降低了重组生肌因子5(Myf5)的表达。然而,干扰LINE-1对FTO和成肌决定蛋白1(MyoD)mRNA水平没有明显影响。免疫荧光分析显示,干扰或过表达LINE-1后,Ki-67蛋白表达没有变化。然而,鬼笔环肽染色显示,干扰LINE-1后牦牛肌肉卫星细胞的肌管融合率提高。

结论

LINE-1和FTO mRNA表达模式的全面图谱确定了牦牛中FTO蛋白与LINE-1 RNA之间的直接相互作用。研究结果表明,FTO过表达促进肌肉卫星细胞分化,而LINE-1负向调节肌管融合。该研究为LINE-1-FTO轴在决定牦牛肌肉卫星细胞命运中的作用提供了基本见解,为未来研究奠定了坚实的理论基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3aa8/11410761/84c423b73867/fvets-11-1448587-g001.jpg

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