Letellier T, Malgat M, Coquet M, Moretto B, Parrot-Roulaud F, Mazat J P
Université Bordeaux II, Neurochimie-CNRS, France.
Pediatr Res. 1992 Jul;32(1):17-22. doi: 10.1203/00006450-199207000-00004.
Respiratory parameters of skeletal muscle were determined in permeabilized muscle fibers by adapting a technique described by Veksler et al. for cardiac fibers (Biochim Biophys Acta, 892:191-196, 1987). This method consists of the permeabilization of muscle fibers by saponin by allowing respiratory substrates and inhibitors to reach the mitochondria. In this way, the mitochondria may be studied inside the fibers as if they were isolated. We have verified, using various techniques, that the mitochondria remain intact during this procedure. This method has been applied to the study of six newborn infants for whom a diagnosis of a mitochondrial defect was suspected. In all cases, the defect was to be found on the permeabilized fibers, and this was confirmed by an enzymatic study. The advantage of this new method, associated with the measurement of the enzymatic activities on a crude homogenate, is to enable a simple and rapid diagnosis on a small amount of sample without damaging the mitochondria during the isolation procedure.
通过采用Veksler等人描述的用于心脏纤维的技术（《生物化学与生物物理学报》，892:191 - 196，1987年），在通透化的肌纤维中测定骨骼肌的呼吸参数。该方法包括通过皂角苷使肌纤维通透化，从而让呼吸底物和抑制剂能够到达线粒体。通过这种方式，可以在纤维内部研究线粒体，就好像它们是被分离出来的一样。我们已经使用各种技术验证，在此过程中线粒体保持完整。该方法已应用于对六名疑似线粒体缺陷的新生儿的研究。在所有病例中，在通透化的纤维上均发现了缺陷，并且这一点通过酶学研究得到了证实。这种新方法与在粗匀浆上测量酶活性相结合的优点是，能够在不破坏线粒体的分离过程中，对少量样本进行简单快速的诊断。
