Cízková Dana, Soukup Tomás, Mokrý Jaroslav
Department of Histology and Embryology, Charles University in Prague, Faculty of Medicine in Hradec Kralove, Prague, Czech Republic.
Cells Tissues Organs. 2009;189(5):338-47. doi: 10.1159/000142161. Epub 2008 Jun 28.
This work provides a comprehensive view of expression and distribution patterns of the intermediate filament nestin in rat regenerating skeletal muscles. Regeneration was induced by heterochronous isotransplantation of extensor digitorum longus (EDL) or soleus muscles from 15-day-old rats into the EDL muscle of adult female inbred Lewis rats. Recipients were sacrificed by anesthetic overdose and the host muscles with the graft were excised after 7-, 16-, 21- and 29-day survival, fixed in 10% formalin solution and immunohistochemically stained. In the regenerating skeletal muscle maximal nestin expression was observed in newly formed myoblasts and myotubes and it decreased in the course of skeletal myofiber development. We demonstrate the phenomenon of a peripheral concentration of nestin expression within the differentiating myofibers called 'lateralization', which may represent a mechanism of nestin elimination during the development of skeletal myofibers. Furthermore, nestin has been detected in newly formed endothelial cells of blood vessels growing into the regenerated area and also in peripheral nerves innervating the graft as well as the host muscle. In conclusion, according to our results, nestin represents an ideal marker for skeletal muscle regeneration, as its expression reflects processes of development, revascularization and reinnervation of new myofibers.
这项研究全面呈现了中间丝蛋白巢蛋白在大鼠再生骨骼肌中的表达及分布模式。通过将15日龄大鼠的趾长伸肌(EDL)或比目鱼肌异时同体移植到成年雌性近交系Lewis大鼠的EDL肌中诱导再生。受体通过过量麻醉处死,在存活7天、16天、21天和29天后,切除带有移植物的宿主肌肉,用10%福尔马林溶液固定并进行免疫组织化学染色。在再生骨骼肌中,巢蛋白在新形成的成肌细胞和肌管中表达最高,并且在骨骼肌纤维发育过程中其表达降低。我们证明了在分化的肌纤维内巢蛋白表达呈外周集中的现象,即“侧向化”,这可能代表了骨骼肌纤维发育过程中巢蛋白消除的一种机制。此外,在长入再生区域的血管的新形成的内皮细胞中以及支配移植物和宿主肌肉的外周神经中也检测到了巢蛋白。总之,根据我们的结果,巢蛋白是骨骼肌再生的理想标志物,因为其表达反映了新肌纤维的发育、血管再生和神经再支配过程。