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Isolation of Human Myoblasts, Assessment of Myogenic Differentiation, and Store-operated Calcium Entry Measurement
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[Regulation of m^(6)A RNA Methylation and Its Effect on Myogenic Differentiation in Murine Myoblasts]

J N Chen1, Y Chen1, Y Y Wei1

  • 1Department of Zoology, College of Life Science, Sichuan Agricultural University, Ya'an, Sichuan, 625014 China.

Molekuliarnaia Biologiia
|June 12, 2019
PubMed

Insights

N-methyladenosine (m6A) RNA methylation positively regulates muscle cell differentiation. METTL3 and betaine increase m6A levels, while cycloleucine decreases them, impacting myogenesis.

Area of Science:

  • Epitranscriptomics
  • Molecular Biology
  • Cellular Metabolism

Background:

  • N^(6)-methyladenosine (m^(6)A) is a crucial epitranscriptomic modification in eukaryotic mRNA.
  • m^(6)A plays significant roles in regulating cellular metabolic processes.
  • The specific role of m^(6)A in myogenic differentiation remains largely unexplored.

Purpose of the Study:

  • To investigate the impact of m^(6)A RNA methylation on the myogenic differentiation of murine myoblasts.
  • To elucidate the regulatory mechanisms of m^(6)A in myogenesis.

Main Methods:

  • Overexpression of METTL3 to alter m^(6)A RNA methylation levels.
  • Utilizing chemical reagents: cycloleucine (Cyc) as an inhibitor and betaine (Bet) as a methyl donor.
  • In vitro experiments on murine myoblasts to assess myogenic differentiation.

Main Results:

  • METTL3 and betaine were found to positively regulate m^(6)A RNA methylation levels.
  • Cycloleucine demonstrated a negative regulatory effect on m^(6)A RNA methylation levels.
  • m^(6)A methylation was shown to positively influence myogenic differentiation in murine myoblasts.

Conclusions:

  • m^(6)A RNA methylation is a positive regulator of myogenic differentiation.
  • These findings offer insights into the molecular mechanisms governing m^(6)A's role in myogenesis.
Keywords:
METTL3m^(6)A RNA methylationmyogenic differentiation

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