Simulated microgravity attenuates myogenic differentiation via epigenetic regulations.
Simulated microgravity attenuates myogenic differentiation via epigenetic regulations.
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DOI：
10.1038/s41526-018-0045-0
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发表时间：
2018
期刊：
影响因子：
5.1
通讯作者：
Yuge L
中科院分区：
文献类型：
--
作者：
Furukawa T;Tanimoto K;Fukazawa T;Imura T;Kawahara Y;Yuge L
肌源性分化的分子机制是相对众所周知的。成肌分化受碱性螺旋-环-螺旋成肌调节转录因子（MRF）的顺序激活调控，生物力学信号在成肌分化的调控中起重要作用。在这项研究中，我们试图确定使用Gravite®的模拟微重力培养是否会影响成肌细胞分化和MRF基因的表达。虽然大鼠成肌细胞，L6细胞分化成肌管在一个孵育期依赖的方式，L6细胞的成肌作用显着减弱模拟微重力（10 - 3G）条件下。实时逆转录聚合酶链反应（RT-PCR）显示，Myog，Myf 6，Mef2c，Des和Ckm的表达在1 G条件下增加的孵育期依赖性的方式，和Myod 1的表达，特别是观察到瞬时增加在早期阶段。然而，在模拟微重力条件下，Myod 1和Myog的表达显着抑制。为了阐明分子机制，L6细胞用5-AzaC处理，并在1 G或10 − 3 G条件下与分化培养基进一步孵育。结果表明，在1 G和10 − 3 G条件下，Myod 1、Myog和的表达水平以及肌管厚度的差异在该实验条件下完全消失。改良的HpaII连接介导PCR（HELP）-分析的微小片段富集表明，在模拟微重力条件下，DNA甲基化状态的动力学变化减弱。这些结果表明，微重力通过控制DNA甲基化来调节肌生成和Myod1表达。由于关键肌肉分化基因的表观遗传调控发生改变，肌肉发育在模拟微重力下受损。来自日本广岛大学的Louis Yuge及其同事在正常重力和微重力条件下培养了大鼠肌肉祖细胞，称为成肌细胞，两种情况下都使用和不使用药物抑制一种酶，该酶参与将甲基标签添加到DNA中以控制基因表达。在没有这种药物的情况下，微重力孵育的细胞以较慢的速度形成肌纤维，并显示出两个关键分化基因表达的缺陷。然而，使用药物后，这些差异消失了。研究结果表明，微重力影响与调节肌肉形成有关的DNA甲基化模式。类似的过程可能发生在与年龄相关的肌肉损失和虚弱的人身上，这突出了未来药物开发的潜在治疗目标。
The molecular mechanisms involved in myogenic differentiation are relatively well-known. Myogenic differentiation is regulated by the sequential activation of the basic helix-loop-helix myogenic regulatory transcription factors (MRFs), and biomechanical signals play an important role in the regulation of myogenesis. In this study, we sought to determine whether simulated microgravity culture using Gravite® may affect myoblast differentiation and expression of MRF genes. Although rat myoblasts, L6 cells were differentiated to myotubes in an incubation period-dependent manner, myogenesis of L6 cells was significantly attenuated under simulated microgravity (10-3G) conditions. Real-time Reverse transcription polymerase chain reaction (RT-PCR) showed that expressions of Myog, Myf6, Mef2c, Des, and Ckm under 1 G conditions increase in an incubation period-dependent manner, and that Myod1 expression was specifically observed to increase transiently in the early phase. However, expressions of Myod1 and Myog were significantly inhibited under simulated microgravity conditions. To clarify the molecular mechanisms, L6 cells were treated with 5-AzaC, and further incubated with differentiation medium under 1 G or 10−3 G conditions. The results showed differences in expression levels of Myod1, Myog, and, as well as those of myotube thickness between 1 G and 10−3 G conditions, completely disappeared in this experimental condition. Modified HpaII tiny fragment enrichment by ligation-mediated PCR (HELP)-assay showed that kinetic changes of DNA methylation status were attenuated in simulated microgravity conditions. These results indicate that microgravity regulates myogenesis and Myod1 expression by controlling DNA methylation. Muscle development is impaired under simulated microgravity because of alterations in the epigenetic regulation of critical muscle differentiation genes. Louis Yuge and colleagues from Hiroshima University, Japan, cultured rat muscle progenitor cells known as myoblasts under normal gravity and microgravity conditions, both with and without a drug that inhibits an enzyme involved in adding methyl tags to DNA to control gene expression. In the absence of this drug, microgravity-incubated cells formed muscle fibers at a slower rate and showed deficits in expression of two key differentiation genes. With the drug, however, these differences disappeared. The findings suggest that microgravity affects patterns of DNA methylation linked to regulating muscle formation. A similar process may be happening in the bodies of people with age-related muscle loss and weakness—highlighting a potential therapeutic target for future drug development.
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影响因子：
9.2
作者：
Montesano A;Luzi L;Senesi P;Terruzzi I
通讯作者：
Terruzzi I
影响因子：
4
作者：
Singh, Kamaleshwar P.;Kumari, Ragini;DuMond, James W.
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影响因子：
3.9
作者：
Pietrangelo, Tiziana;Puglielli, Cristina;Fulle, Stefania
通讯作者：
Fulle, Stefania
影响因子：
4.3
作者：
Laker RC;Ryall JG
通讯作者：
Ryall JG
影响因子：
29
作者：
Barres, Romain;Yan, Jie;Zierath, Juleen R.
通讯作者：
Zierath, Juleen R.