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Higher PLIN5 but not PLIN3 content in isolated skeletal muscle mitochondria following acute in vivo contraction in rat hindlimb

机译:大鼠后肢急性体内收缩后分离的骨骼肌线粒体中pLIN5含量较高但pLIN3含量较高

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摘要

Contraction-mediated lipolysis increases the association of lipid droplets and mitochondria, indicating an important role in the passage of fatty acids from lipid droplets to mitochondria in skeletal muscle. PLIN3 and PLIN5 are of particular interest to the lipid droplet–mitochondria interaction because PLIN3 is able to move about within cells and PLIN5 associates with skeletalmuscle mitochondria. This study primarily investigated: 1) if PLIN3 is detected in skeletal muscle mitochondrial fraction; and 2) if mitochondrial protein content of PLIN3 and/or PLIN5 changes following stimulated contraction. A secondary aim was to determine if PLIN3 and PLIN5 associate and whether this changes following contraction. Male Long Evans rats (n = 21;age, 52 days; weight = 317 6 g) underwent 30 min of hindlimb stimulation (10 msec impulses, 100 Hz/3 sec at 10–20 V; train duration 100 msec). Contraction induced a ~50% reduction in intramuscular lipid content measured by oil red-O staining of red gastrocnemius muscle. Mitochondria were isolated from red gastrocnemius muscle by differential centrifugation and proteinswere detected by western blotting. Mitochondrial PLIN5 content was ~1.6-fold higher following 30 min of contraction and PLIN3 content was detected in the mitochondrial fraction, and unchanged following contraction. An association between PLIN3 and PLIN5 was observed and remained unaltered following contraction. PLIN5 may play a role in mitochondria during lipolysis, which is consistent with a role in facilitating/regulating mitochondrial fatty acid oxidation. PLIN3 and PLIN5 may be working together on the lipid droplet and mitochondria during contraction-induced lipolysis.
机译:收缩介导的脂解作用增加了脂滴与线粒体的缔合,表明在脂肪酸从脂滴到骨骼肌线粒体的通道中起着重要作用。 PLIN3和PLIN5对于脂滴与线粒体的相互作用特别感兴趣,因为PLIN3能够在细胞内移动,并且PLIN5与骨骼肌线粒体相关。这项研究主要调查:1)是否在骨骼肌线粒体部分检测到PLIN3; 2)PLIN3和/或PLIN5的线粒体蛋白含量在刺激后是否发生变化。第二个目的是确定PLIN3和PLIN5是否缔合,以及收缩后是否改变。雄性Long Evans大鼠(n = 21;年龄52天;体重= 317 6 g)进行了30分钟的后肢刺激(10毫秒脉冲,在10–20 V时为100 Hz / 3秒;训练持续时间为100毫秒)。通过腓肠肌红色油的油红-O染色测量,收缩引起的肌内脂质含量降低了约50%。通过差异离心法从腓肠肌的红色肌肉中分离出线粒体,并通过蛋白质印迹法检测蛋白质。线粒体收缩后30分钟,线粒体PLIN5含量高约1.6倍,线粒体级分中检测到PLIN3含量,收缩后未改变。观察到PLIN3和PLIN5之间的关联,并且在收缩后保持不变。 PLIN5可能在脂解过程中在线粒体中发挥作用,这与促进/调节线粒体脂肪酸氧化的作用一致。在收缩诱导的脂解过程中,PLIN3和PLIN5可能共同作用于脂质滴和线粒体。

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