Cardasis C A, Cooper G W
J Exp Zool. 1975 Mar;191(3):333-46. doi: 10.1002/jez.1401910304.
A method is described for isolating intact, individual skeletal muscle fibers from glutaraldehyde fixed muscle. This method was conceived to climinate the many limitations of determining muscle nuclear numbers in histological cross section. Glutaraldehyde fixed fibers are isolated by dissection while in a solution of low concentration guanidine in a borate buffer at high pH. Electron miscroscopy demonstrates that single fibers, isolated in this manner, are free of their microvasculature and connective tissue. Their basal laminas and the structures within them, including their satellite cells, are preserved. This method is employed to determine whether changes in nuclear numbers occur within the basal lamina of individual muscle cells from 1 to 28 days following denervation of mouse gastrocnemius muscle. The total number of nuclei located within the basal lamina of individual muscle fibers (i.e. muscle and satellite cell nuclei) does not change after denervation. This rules out the possibility that additional nuclei are arising from an influx of cells outside the basal lamina or by mitotic division of nuclei within the basal lamina. However, the possibility of a change in the ratio of satellite cell nuclei, to muscle cell nuclei, is not eliminated. Other possible applications of this isolation method are discussed.
本文描述了一种从戊二醛固定的肌肉中分离完整的单个骨骼肌纤维的方法。构思该方法是为了消除在组织学横切面上确定肌肉核数量时的诸多限制。戊二醛固定的纤维在高pH值的硼酸盐缓冲液中的低浓度胍溶液中通过解剖进行分离。电子显微镜显示，以这种方式分离的单根纤维没有其微血管和结缔组织。它们的基膜及其内部结构，包括卫星细胞，都得以保留。该方法用于确定小鼠腓肠肌去神经支配后1至28天内，单个肌肉细胞基膜内的核数量是否发生变化。去神经支配后，单个肌肉纤维基膜内的核总数（即肌核和卫星细胞核）没有变化。这排除了额外的核是由基膜外细胞流入或基膜内核的有丝分裂产生的可能性。然而，卫星细胞核与肌细胞核比例变化的可能性并未排除。还讨论了这种分离方法的其他可能应用。
