Csukly Kristina, Ascah Alexis, Matas Jimmy, Gardiner Phillip F, Fontaine Eric, Burelle Yan
Département de kinésiologie, Université de Montréal, P.O. Box 6128 Centre-Ville, Montreal, Quebec, Canada, H3C 3J7.
J Physiol. 2006 Jul 1;574(Pt 1):319-27. doi: 10.1113/jphysiol.2006.109702. Epub 2006 May 4.
Loss of neural input to skeletal muscle fibres induces atrophy and degeneration with evidence of mitochondria-mediated cell death. However, the effect of denervation on the permeability transition pore (PTP), a mitochondrial protein complex implicated in cell death, is uncertain. In the present study, the impact of 21 days of denervation on the sensitivity of the PTP to Ca2+-induced opening was studied in isolated muscle mitochondria. Muscle denervation increased the sensitivity to Ca2+-induced opening of the PTP, as indicated by a significant decrease in calcium retention capacity (CRC: 111 +/- 12 versus 475 +/- 33 nmol (mg protein)(-1) for denervated and sham, respectively). This phenomenon was partly attributable to in vivo mitochondrial and whole muscle Ca2+ overload. Cyclosporin A, which inhibits PTP opening by binding to cyclophilin D (CypD), was significantly more potent in mitochondria from denervated muscle and restored CRC to the level observed in mitochondria from sham-operated muscles. In contrast, the CypD independent inhibitor trifluoperazine was equally effective at inhibiting PTP opening in sham and denervated animals and did not correct the difference in CRC between groups. This phenomenon was associated with a significant increase in the content of the PTP regulating protein CypD relative to several mitochondrial marker proteins. Together, these results indicate that Ca2+ overload in vivo and an altered expression of CypD could predispose mitochondria to permeability transition in denervated muscles.
骨骼肌纤维的神经输入丧失会导致萎缩和退化,并伴有线粒体介导的细胞死亡迹象。然而,去神经支配对通透性转换孔(PTP)(一种与细胞死亡有关的线粒体蛋白复合物)的影响尚不确定。在本研究中,在分离的肌肉线粒体中研究了21天去神经支配对PTP对Ca2+诱导开放的敏感性的影响。肌肉去神经支配增加了PTP对Ca2+诱导开放的敏感性,这表现为钙保留能力(CRC)显著降低(去神经支配组和假手术组的CRC分别为111±12和475±33 nmol(mg蛋白)-1)。这种现象部分归因于体内线粒体和全肌Ca2+过载。环孢素A通过与亲环蛋白D(CypD)结合来抑制PTP开放,在去神经支配肌肉的线粒体中其效力显著更强,并使CRC恢复到假手术肌肉线粒体中观察到的水平。相反,不依赖CypD的抑制剂三氟拉嗪在假手术组和去神经支配组动物中抑制PTP开放的效果相同,并且没有纠正两组之间CRC的差异。这种现象与PTP调节蛋白CypD相对于几种线粒体标记蛋白的含量显著增加有关。总之,这些结果表明体内Ca2+过载和CypD表达改变可能使去神经支配肌肉中的线粒体易于发生通透性转换。