Network distribution of mitochondria and lipid droplets in human muscle fibres
Network distribution of mitochondria and lipid droplets in human muscle fibres
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DOI：
10.1007/s00418-007-0349-8
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发表时间：
2008-01-01
影响因子：
2.3
通讯作者：
Wagenmakers, Anton J. M.
中科院分区：
文献类型：
--
作者：
Shaw, Christopher S.;Jones, David A.;Wagenmakers, Anton J. M.
本研究的目的是开发一种荧光染色组合，通过传统和共聚焦显微镜观察人类骨骼肌纤维中的线粒体和脂滴（肌细胞内脂质或 IMCL）网络。肌肉活检取自三名瘦弱、健康且身体活跃的男性受试者的股外侧肌。使用针对三种线粒体蛋白的抗体对冷冻肌肉切片进行线粒体染色；孔蛋白、细胞色素c氧化酶（COX）和NADH-泛醇氧化还原酶以及中性脂质用油红O染色。抗COX染色产生的图像具有最强的荧光信号和最高分辨率的线粒体网络，并且该染色剂成功地与抗I型纤维肌球蛋白的抗体结合。在氧化型 I 纤维中观察到肌节内线粒体的高度组织化矩阵排列（在 I 带成对）。肌膜下区域线粒体密度最高。抗 COX 染色与油红 O 结合表明，在 I 型纤维中，脂滴主要位于线粒体之间的空间。
The objective of the present study was to develop a combination of fluorescent stains that would allow visualisation of the network of mitochondria and lipid droplets (intramyocellular lipids or IMCL) in human skeletal muscle fibres by means of conventional and confocal microscopy. Muscle biopsies were taken from the vastus lateralis of three lean, healthy and physically active male subjects. Frozen muscle sections were stained for mitochondria using antibodies against three mitochondrial proteins; porin, cytochrome c oxidase (COX) and NADH-ubiquinol oxidoreductase and neutral lipids were stained with oil red O. Anti-COX staining produced images with the strongest fluorescence signal and the highest resolution of the mitochondrial network and this stain was successfully combined with the antibody against type I fibre myosin. A highly organised matrix arrangement of mitochondria within the sarcomeres (in pairs at the I-band) was observed in the oxidative type I fibres. The density of mitochondria was the highest in the subsarcolemmal region. Anti-COX staining was combined with oil red O demonstrating that in type I fibres lipid droplets are mainly located in the space between the mitochondria.