Transcriptional Regulation of Muscle-Specific Genes during Myoblast Differentiation

• Published in: Biochemical and biophysical research communications 1998-05, Vol.246 (1), p.287-292
• Main Authors: Shimokawa, Teruhiko, Kato, Miyuki, Ezaki, Osamu, Hashimoto, Seiichi
• Format: Article
• Language: English
• Subjects: Animals; Base Sequence; Cell Differentiation - genetics; Cell Line; DNA Primers - genetics; DNA, Complementary - genetics; Gene Expression Regulation, Developmental; Kinetics; Mice; Molecular Sequence Data; Muscle Proteins - genetics; Muscles - cytology; Muscles - metabolism; Myogenic Regulatory Factors - genetics; Polymerase Chain Reaction - methods; Polymerase Chain Reaction - statistics & numerical data; RNA, Messenger - genetics; RNA, Messenger - metabolism; Sensitivity and Specificity
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1006/bbrc.1998.8600

A rapid, highly sensitive method to determine the mRNA level of muscle-specific markers using TaqMan PCR analysis was developed and used to study sequential gene regulation of myoblasts during induced differentiation of C2C12cells. mRNA levels of muscle regulatory factor (MRF) myogenin, α-actin, thermoregulatory uncoupling protein UCP2 and glucose transporter isotype glut4 increased rapidly during early stage differentiation. In contrast, myf5, β-actin, UCP1 and glut1 mRNA levels gradually decreased during 8 days. However, the mRNA level of other genes such as MyoD, glyceraldehyde-3-phosphate dehydrogenase and hexokinase II changed only slightly in comparison. Muscle-specific uncoupling protein UCP3 mRNA was detected during differentiation and increased rapidly within 8 days. These data clearly show the sequential and the differential regulation of muscle-specific genes in C2C12cells during multinucleate myotube formation.