Abstract
Myofibers have characteristic membrane compartments in their cytoplasm and sarcolemma, such as the sarcoplasmic reticulum, T-tubules, neuromuscular junction, and myotendinous junction. Little is known about the vesicular transport that is believed to mediate the development of these membrane compartments. We determined the locations of organelles in differentiating myotubes. Electron microscopic observation of a whole myotube revealed the arrangement of Golgi apparatus, rough endoplasmic reticulum, autolysosomes, mitochondria, and smooth endoplasmic reticulum from the perinuclear region toward the end of myotubes and the existence of a large number of vesicles near the ends of myotubes. Vesicles in myotubes were further characterized using immunofluorescence microscopy to analyze expression and localization of vesicle-associated membrane proteins (VAMPs). VAMPs are a family of seven proteins that regulate post-Golgi vesicular transport via the fusion of vesicles to the target membranes. Myotubes express five VAMPs in total. Vesicles with VAMP2, VAMP3, or VAMP5 were found near the ends of the myotubes. Some of these vesicles are also positive for caveolin-3, suggesting their participation in the development of T-tubules. Our morphological analyses revealed the characteristic arrangement of organelles in myotubes and the existence of transport vesicles near the ends of the myotubes.
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Acknowledgments
We thank Ms. Harumi Matsuda and Mr. Yoshihiro Morimura (Department of Anatomy, Gunma University Graduate School of Medicine) for both technical and secretarial assistance and Mr. Hiroyuki Seo and Dr. Touko Hirano (Laboratory for Analytical Instruments, Education and Research Support Center, Gunma University Graduate School of Medicine) for their support for the EM preparation and observation. We also thank Mr. Hisao Yajima and Mr. Ryota Koyama (Gunma University School of Medicine) for cooperating in technical matters as a part of Practical Course on Basic Medical Science. This work was supported by MEXT KAKENHI Grant Numbers 21790175, 23590230, and 25860138.
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Tajika, Y., Takahashi, M., Khairani, A.F. et al. Vesicular transport system in myotubes: ultrastructural study and signposting with vesicle-associated membrane proteins. Histochem Cell Biol 141, 441–454 (2014). https://doi.org/10.1007/s00418-013-1164-z
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DOI: https://doi.org/10.1007/s00418-013-1164-z