Nitric oxide drives embryonic myogenesis in chicken through the upregulation of myogenic differentiation factors

Exp Cell Res. 2014 Jan 15;320(2):269-80. doi: 10.1016/j.yexcr.2013.11.006. Epub 2013 Nov 15.

Abstract

The muscle-specific variant of neuronal nitric oxide (NO) synthase (NOS-I), is developmentally regulated in mouse suggesting a role of NO during myogenesis. In chick embryo, a good model of development, we found that the expression of NOS-I is up-regulated, but only in the early phase of development. Through a pharmacological intervention in ovo we found that NO signalling plays a relevant role during embryonic development. The inhibition of NOS-I decreased the growth of embryo, in particular of muscle tissue, while the restoring of physiological NO levels, via administration of a NO donor, reversed this effect. We found a selective action of NO, produced by NOS-I, on regulatory factors involved in myogenic differentiation in the early phase of chick embryo development: inhibition of NO generation leads to a decreased expression of the Myocyte enhancer factor 2a (Mef2a), Mef2c, Myogenin and Myosin, which was reversed by the administration of a NO donor. NO had no effects on Myf5 and MyoD, the myogenic regulatory factors necessary for myogenic determination. The action of NO on the myogenic regulatory factors was mediated via generation of cyclic GMP (cGMP) and activation of the cGMP-dependent protein kinase G (PKG). Finally we found in myoblasts in vitro that the activation of Mef2c was the key event mediating the NO-induced modulation of myogenesis. Our results identify NO produced by NOS-I as a key messenger in the early phase of embryonic development of chicken, acting as a critical determinant of myogenesis through its physiological cGMP/PKG pathway.

Keywords: 1-[N-(2-aminoethyl)–N-(2-ammonioethyl)amino]diazen-1-ium-1,2-diolate, DMEM, Dulbecco's Modified Eagle's Medium; 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one; Chick embryo; DETA; FBS; GAPDH; HH; HRP; Hamburger and Hamilton; MRFs; Mef2; Mef2c; Murine myoblasts; Muscle development; Myocyte enhancer factor 2; N(ω)-propyl-l-arginine; NO; NO synthases; NOS; NOS-I; NPLA; ODQ; PBS; PKG; SDS; cGMP; cGMP-dependent protein kinase; cGMP/PKG; cyclic GMP; foetal bovine serum; glucose 6 phosphate dehydrogenase; horseradish-peroxidase; myogenic regulatory factors; neuronal NOS; nitric oxide; phosphate buffered saline; sGC; sodium dodecyl sulphate; soluble guanylate cyclase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Differentiation / drug effects
  • Cell Differentiation / genetics
  • Cells, Cultured
  • Chick Embryo
  • Chickens / genetics
  • Chickens / metabolism
  • Gene Expression Regulation, Developmental / drug effects*
  • Humans
  • Mice
  • Muscle Development / drug effects*
  • Muscle Development / genetics*
  • Myogenic Regulatory Factors / genetics*
  • Myogenic Regulatory Factors / metabolism
  • Nitric Oxide / pharmacology*
  • Nitric Oxide Synthase Type I / genetics
  • Nitric Oxide Synthase Type I / metabolism
  • Up-Regulation / drug effects

Substances

  • Myogenic Regulatory Factors
  • Nitric Oxide
  • Nitric Oxide Synthase Type I