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

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• 作者：Denise Cazzato ; Emma Assi ; Claudia Moscheni ; Silvia Brunelli ; Clara De Palma ; Davide Cervia ; Cristiana Perrotta ; Emilio Clementi
• 关键词：cGMP ; cyclic GMP ; DETA ; 1-[N-(2-aminoethyl)&ndash ; N-(2-ammonioethyl)amino]diazen-1-ium-1 ; 2-diolate ; DMEM ; Dulbecco's Modified Eagle's Medium ; FBS ; foetal bovine serum ; GAPDH ; glucose 6 phosphate dehydrogenase ; HH ; Hamburger and Hamilton ; HRP ; horseradish-peroxidase ; Mef2 ; Myocyte enhancer factor 2 ; MRFs ; myogenic regulatory factors ; NO ; nitric oxide ; NOS ; NO synthases ; NOS-I ; neuronal NOS ; NPLA ; N蠅-propyl-l-arginine ; ODQ ; 1H-[1 ; 2 ; 4]oxadiazolo[4 ; 3-a]quinoxalin-1-one ; PBS ; phosphate buffered sali
• 刊名：Experimental Cell Research
• 出版年：15 January, 2014
• 年：2014
• 卷：320
• 期：2
• 页码：269-280
• 全文大小：3675 K

文摘

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.