miR-196a 和 miR-196b 在斑马鱼运动功能中的作用。

Roles of miR-196a and miR-196b in Zebrafish Motor Function.

机构信息

Laboratory of Molecular and Statistical Genetics, College of Life Sciences, Hunan Normal University, Changsha 410081, China.

College of Physical Education and Health, Hunan University of Technology and Business, Changsha 410025, China.

出版信息

Biomolecules. 2023 Mar 17;13(3):554. doi: 10.3390/biom13030554.

DOI:10.3390/biom13030554

PMID:36979489

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10046552/

Abstract

BACKGROUND

The exertion of motor function depends on various tissues, such as bones and muscles. miR-196 has been widely studied in cancer and other fields, but its effect on bone and skeletal muscle is rarely reported. In order to explore the role of miR-196 family in bone and skeletal muscle, we used the previously successfully constructed miR-196a-1 and miR-196b gene knockout zebrafish animal models for research.

METHODS

The behavioral trajectories of zebrafish from 4 days post-fertilization (dpf) to 7 dpf were detected to analyze the effect of miR-196a-1 and miR-196b on motor ability. Hematoxylin-eosin (HE) staining and transmission electron microscopy (TEM) were used to detect the dorsal muscle tissue of zebrafish. The bone tissue of zebrafish was detected by microcomputed tomography (micro-CT). Real-time PCR was used to detect the expression levels of related genes, including vcp, dpm1, acta1b, mylpfb, col1a1a, bmp8a, gdf6a, and fgfr3.

RESULTS

The behavioral test showed that the total behavioral trajectory, movement time, and movement speed of zebrafish larvae were decreased in the miR-196a-1 and miR-196b gene knockout lines. Muscle tissue analysis showed that the structure of muscle fibers in the zebrafish lacking miR-196a-1 and miR-196b was abnormal and was characterized by vacuolar degeneration of muscle fibers, intranuclear migration, melanin deposition, and inflammatory cell infiltration. Bone CT examination revealed decreased bone mineral density and trabecular bone number. The real-time PCR results showed that the expression levels of vcp, dpm1, gdf6a, fgfr3, and col1a1a were decreased in the miR-196b gene knockout group. The expression levels of dpm1, acta1b, mylpfb, gdf6a, and col1a1a were decreased, and the expression level of fgfr3 was increased in the miR-196b gene knockout group compared with the wild-type group.

CONCLUSIONS

miR-196a-1 and miR-196b play an important role in muscle fiber structure, bone mineral density, and bone trabecular quantity by affecting the expression of vcp, dpm1, acta1b, mylpfb, gdf6a, fgfr3, and col1a1a and then affect the function of the motor system.

摘要

背景

运动功能的发挥依赖于骨骼和肌肉等多种组织。miR-196 在癌症等领域已有广泛研究，但对骨骼和骨骼肌的作用鲜有报道。为了探讨 miR-196 家族在骨骼和骨骼肌中的作用，我们利用前期成功构建的 miR-196a-1 和 miR-196b 基因敲除斑马鱼动物模型进行研究。

方法

检测 4 日龄至 7 日龄斑马鱼的行为轨迹，分析 miR-196a-1 和 miR-196b 对运动能力的影响。采用苏木精-伊红（HE）染色和透射电镜（TEM）观察斑马鱼背部肌肉组织。采用微计算机断层扫描（micro-CT）检测斑马鱼的骨骼组织。实时 PCR 检测 vcp、dpm1、acta1b、mylpfb、col1a1a、bmp8a、gdf6a 和 fgfr3 等相关基因的表达水平。

结果

行为学测试显示，miR-196a-1 和 miR-196b 基因敲除系斑马鱼幼虫的总行为轨迹、运动时间和运动速度均降低。肌肉组织分析显示，miR-196a-1 和 miR-196b 缺失的斑马鱼肌肉纤维结构异常，表现为肌纤维空泡变性、核内迁移、黑色素沉积和炎性细胞浸润。骨 CT 检查显示骨密度和小梁骨数量减少。实时 PCR 结果显示，miR-196b 基因敲除组 vcp、dpm1、gdf6a、fgfr3 和 col1a1a 的表达水平降低。与野生型相比，miR-196b 基因敲除组 dpm1、acta1b、mylpfb、gdf6a 和 col1a1a 的表达水平降低，fgfr3 的表达水平升高。