TY - JOUR
T1 - Expression of neurofilament proteins in proliferating C2C12 mouse skeletal muscle cells
AU - Masami, Abe
AU - Saitoh, Osamu
AU - Nakata, Hiroyasu
AU - Yoda, Akinori
AU - Matsuda, Ryoichi
N1 - Funding Information:
We thank Drs. Takashi Obinata and Takeshi Endo at Department of Biology, Chiba University, for their generous gifts of NT302, MF20, C2C12 cells, and pTnT-15. This study was supported by research Grants 5A-1, from the National Center of Neurology and Psychiatry of the Ministry of Health and Welfare, Japan; 06213214 and 08233102, for scienti®c research on priority areas, and 06804054 and 088741194, from the Ministry of Education, Science and Culture, Japan, and the Fugaku Trust for Medicinal Research, Tokyo, Japan, to R.M.; and 951048 from Kanagawa Academy of Science and Technology to O.S.
PY - 1996/11/25
Y1 - 1996/11/25
N2 - Expression of neurofilament proteins, NF140K, NF68K, and NF200K, in C2C12 mouse skeletal muscle cells was studied. Immunofluorescence and immunoblot analyses revealed that NF140K was expressed in proliferating C2C12 cells and its localization was similar to desmin, a muscle-specific intermediate filament protein. NF140K became undetectable in C2C12 cells as muscle cell differentiation proceeded. Reverse transcription-polymerase chain reaction (RT-PCR) and Northern blot analyses confirmed the expression of NF140K, NF68K, and NF200K in proliferating C2C12 cells. Sequences of the RT- PCR products of NF140K and NF68K were identical to that of authentic mouse NF140K and NF68K, respectively. The NF140K and NF68K mRNA were down-regulated during myogenesis in contrast to the up-regulation of mRNA encoding troponin- T. Furthermore, subcloned C2C12 cells, which express NF140K at a higher level, exhibited retarded myogenesis, i.e., delayed onset of myosin heavy chain synthesis and myoblast fusion. These results suggest that neurofilament proteins may play an inhibitory role in commitment of muscle cell differentiation. The presence of neurofilament proteins in skeletal muscle cells indicates that desmin, vimentin, and neurofilament proteins can be expressed in a single muscle cell.
AB - Expression of neurofilament proteins, NF140K, NF68K, and NF200K, in C2C12 mouse skeletal muscle cells was studied. Immunofluorescence and immunoblot analyses revealed that NF140K was expressed in proliferating C2C12 cells and its localization was similar to desmin, a muscle-specific intermediate filament protein. NF140K became undetectable in C2C12 cells as muscle cell differentiation proceeded. Reverse transcription-polymerase chain reaction (RT-PCR) and Northern blot analyses confirmed the expression of NF140K, NF68K, and NF200K in proliferating C2C12 cells. Sequences of the RT- PCR products of NF140K and NF68K were identical to that of authentic mouse NF140K and NF68K, respectively. The NF140K and NF68K mRNA were down-regulated during myogenesis in contrast to the up-regulation of mRNA encoding troponin- T. Furthermore, subcloned C2C12 cells, which express NF140K at a higher level, exhibited retarded myogenesis, i.e., delayed onset of myosin heavy chain synthesis and myoblast fusion. These results suggest that neurofilament proteins may play an inhibitory role in commitment of muscle cell differentiation. The presence of neurofilament proteins in skeletal muscle cells indicates that desmin, vimentin, and neurofilament proteins can be expressed in a single muscle cell.
UR - https://www.scopus.com/pages/publications/17444439960
U2 - 10.1006/excr.1996.0342
DO - 10.1006/excr.1996.0342
M3 - Article
C2 - 8940248
AN - SCOPUS:17444439960
SN - 0014-4827
VL - 229
SP - 48
EP - 59
JO - Experimental Cell Research
JF - Experimental Cell Research
IS - 1
ER -