Transmission Electron Microscopy for Analysis of Mitochondria in Mouse Skeletal Muscle
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Author:
Date:
2018-05-20
[Abstract] Skeletal muscle is the most abundant tissue in the human body and regulates a variety of functions including locomotion and whole-body metabolism. Skeletal muscle has a plethora of mitochondria, the organelles that are essential for aerobic generation of ATP which provides the chemical energy to fuel vital functions such as contraction. The number of mitochondria in skeletal muscle and their function decline with normal aging and in various neuromuscular diseases and in catabolic conditions such as cancer, starvation, denervation, and immobilization. Moreover, compromised mitochondrial function is also associated with metabolic disorders including type 2 diabetes mellitus. It is now clear that maintaining mitochondrial content and function in skeletal muscle is vital for sustained health ...
[摘要] 骨骼肌是人体中含量最丰富的组织,可调节各种功能,包括运动和全身代谢。骨骼肌有很多线粒体,这是ATP好氧生成所必需的细胞器,它提供化学能量来促进收缩等重要功能。骨骼肌中线粒体的数量及其功能随着正常衰老和各种神经肌肉疾病以及癌症,饥饿,去神经支配和固定等分解代谢条件而下降。此外,受损的线粒体功能也与包括2型糖尿病在内的代谢紊乱有关。现在清楚的是维持骨骼肌中的线粒体含量和功能对于整个寿命期间的持续健康是至关重要的。虽然有许多染色方法可用于研究线粒体,但透射电子显微镜(TEM)仍然是研究骨骼肌中线粒体结构和健康的最重要方法。它提供关于线粒体含量,嵴密度,组织,自噬体形成以及在各种疾病状况中经常观察到的任何其他异常的关键信息。在本文中,我们描述了一个详细的协议样本制备和透射电镜分析小鼠骨骼肌线粒体。
【背景】骨骼肌是一种高度塑性的组织,经过响应一些细胞外信号的形态和代谢适应性。包括抵抗或耐力运动在内的许多干扰刺激线粒体生物发生,导致增加的代谢能力和抵抗疲劳(Li等人,2008; Sandri,2008)。相反,在衰老期间,不活动,以及在许多分解代谢疾病状态下,骨骼肌线粒体数量和功能下降,导致易疲劳性和胰岛素抵抗增加(Sandri,2008)。功能失调的线粒体的累积也可能导致进行性活性氧物质诱导的损伤,从而进一步损害骨骼肌中的氧化能力(Bonnard等人,2008)。 ...
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Isolation, Culturing, and Differentiation of Primary Myoblasts from Skeletal Muscle of Adult Mice
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Author:
Date:
2017-05-05
[Abstract] Myogenesis is a multi-step process that leads to the formation of skeletal muscle during embryonic development and repair of injured myofibers. In this process, myoblasts are the main effector cell type which fuse with each other or to injured myofibers leading to the formation of new myofibers or regeneration of skeletal muscle in adults. Many steps of myogenesis can be recapitulated through in vitro differentiation of myoblasts into myotubes. Most laboratories use immortalized myogenic cells lines that also differentiate into myotubes. Although these cell lines have been found quite useful to delineating the regulatory mechanisms of myogenesis, they often show a great degree of variability depending on the origin of the cells and culture conditions. Primary myoblasts have been ...
[摘要] 造血是一种多步骤过程,导致在损伤的肌纤维的胚胎发育和修复期间骨骼肌的形成。在这个过程中,成肌细胞是主要的效应细胞类型,彼此融合或损伤肌纤维,导致新成肌纤维的形成或成年人骨骼肌的再生。通过体外成骨细胞分化成肌管可以概括出许多发生肌肉发育的步骤。大多数实验室使用也分化成肌管的永生化肌原细胞系。虽然已经发现这些细胞系对于描绘造血的调节机制非常有用,但是它们通常依赖于细胞的来源和培养条件而显示出很大的变异性。原代成肌细胞被认为是体外研究肌生成的最生理学相关模型。然而,由于成体骨骼肌的丰度低,原代成肌细胞的分离在技术上是有挑战性的。在本文中,我们描述了一种用于从小鼠的成年骨骼肌分离原代成肌细胞的改进方案。我们还描述了其培养和分化成肌管的方法。
背景 造血是一个复杂而高度协调的过程,其涉及多潜能中胚层细胞的测定,以产生成肌细胞,成肌细胞从细胞周期中排出,以及它们最终分化为骨骼肌纤维。 Myogen-5,MyoD,myogenin和MRF4的基因螺旋 - 环 - 螺旋转录因子的一组基因调控因子(MRFs)的顺序表达调控。 Myf-5和MyoD是成肌细胞形成,增殖和存活所需的主要MRFs,而其他MRF(如肌细胞生成素和MRF-4)在肌发生过程中起作用迟发,激活收缩蛋白和其他结构和代谢蛋白的基因表达(白金汉,2003; ...
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