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Reorganization of microtubule nucleation during muscle differentiation.

机译：肌肉分化过程中微管成核的重组。

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Skeletal muscle differentiation involves a complete reorganization of the microtubule network. Nearly 20 years ago, Tassin et al. [1985: J Cell Biol 100:35-46] suggested a mechanism for this reorganization by showing a redistribution of the microtubule organizing center from the centrosome to the nuclear membrane. Little progress has been made since. It is still not clear whether centrosomal proteins are redistributed together, whether microtubules are nucleated at the nuclear membrane or transported there post-nucleation, and whether gamma-tubulin (gammatub) remains necessary for nucleation in myotubes. To investigate these questions, we have examined the redistribution of the centrosomal proteins pericentrin (PC), gammatub, and ninein in the C2 muscle cell line. Immunofluorescence of differentiated myotubes shows PC along the nuclear membrane whereas gammatub is only detected there after pre-fixation detergent extraction. After expression of a GFP-tagged gammatub, we observe a weak fluorescence alongthe nuclear membrane, confirming the presence of gammatub at a low concentration relative to PC. Microinjection of anti-gammatub antibodies into myotubes blocks microtubule growth from both nuclear membranes and centrosomal sites. The centrosomal microtubule-anchoring protein, ninein, is found at the nuclear membrane as well and its distribution appears independent of microtubule integrity. We conclude that centrosomal proteins are redistributed independently during muscle differentiation, to sites that nucleate microtubules both along the nuclear membranes and through the cytoplasm.
机译：骨骼肌分化涉及微管网络的完全重组。近20年前，Tassin等人。 [1985：J Cell Biol 100：35-46]通过显示微管组织中心从中心体到核膜的重新分布，提出了这种重组的机制。此后，进展甚微。尚不清楚中心体蛋白是否重新分布在一起，微管是否在核膜上成核或在成核后转运到核膜上，以及是否仍需要γ-微管蛋白（γ-tubulin）在肌管中成核。为了调查这些问题，我们检查了C2肌肉细胞系中中心体蛋白高蛋白（PC），γ管和9in的重新分布。分化后的肌管的免疫荧光显示沿核膜有PC，而γ管仅在固定剂去污剂提取后才能在那里检测到。表达GFP标记的gammatub后，我们观察到沿核膜的荧光较弱，证实了相对于PC低浓度的gammatub的存在。将抗gammatub抗体显微注射到肌管中可阻止微管从核膜和中心体位点生长。在核膜上也发现了锚定在中心的微管蛋白，蛋白ininin，其分布与微管的完整性无关。我们得出的结论是，中心体蛋白在肌肉分化过程中独立地重新分布到沿核膜和通过细胞质使微管成核的位点。

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