Nitric oxide facilitates NFAT-dependent transcription in mouse myotubes
Nitric oxide facilitates NFAT-dependent transcription in mouse myotubes
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DOI：
10.1152/ajpcell.00523.2007
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发表时间：
2008-04-01
影响因子：
5.5
通讯作者：
Criswell, David S.
中科院分区：
文献类型：
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作者：
Drenning, Jason A.;Lira, Vitor A.;Criswell, David S.
骨骼肌细胞内钙瞬变通过钙调磷酸酶及其效应核因子活化t细胞（NFAT）启动表型适应。此外，内源性一氧化氮（NO）的产生通过钙钙调素依赖的NO合成酶（NOS）参与骨骼肌表型可塑性。在这里，我们提供的证据表明，NO增强钙依赖性核积累和NFAT的转录活性，并诱导C2C12肌管中糖原合成酶激酶- 3 β (GSK- 3 β)的磷酸化。钙离子载体A23187 （1 μ M持续9小时）或thapsignargin （2 μ M持续4小时）在肌管中瞬时转染NFAT依赖性报告质粒（pNFAT- luc, Stratagene）后，NFAT转录活性分别提高了7倍和4倍。与NOS-抑制剂N-G-硝基- L-精氨酸甲酯（L- NAME; 5 mM）或鸟苷酸环化酶抑制剂1H[1,2,4]恶二唑[4,3- a]喹啉- 1- 1 （ODQ; 10 μ M）共处理可阻止钙对NFAT活性的影响。NO供体二乙基三胺- nono （DETA- NO; 10 μ M）增强了A23187对NFAT依赖性转录的作用。同样，A23187 （0.4 μ M，作用4小时）引起细胞核NFAT的积累，并增加GSK- 3 β的磷酸化（即失活），而与L- NAME或ODQ共处理可抑制这些反应。最后，NO供体3-（2-羟基- 2-亚硝基- 1-丙基肼基）- 1-丙胺（PAPA-NO； 1 μ M作用1小时）以依赖于鸟苷酸环化酶活性的方式增加GSK- 3 β的磷酸化。我们得出结论，NOS活性介导钙诱导的GSK- 3 β磷酸化和肌管中NFAT依赖性转录的激活。此外，一氧化氮的这些作用是鸟苷酸环化酶依赖的。
Intracellular calcium transients in skeletal muscle cells initiate phenotypic adaptations via activation of calcineurin and its effector nuclear factor of activated t- cells ( NFAT). Furthermore, endogenous production of nitric oxide ( NO) via calcium- calmodulin- dependent NO synthase ( NOS) is involved in skeletal muscle phenotypic plasticity. Here, we provide evidence that NO enhances calcium- dependent nuclear accumulation and transcriptional activity of NFAT and induces phosphorylation of glycogen synthase kinase- 3 beta ( GSK- 3 beta) in C2C12 myotubes. The calcium ionophore A23187 ( 1 mu M for 9 h) or thapsigargin ( 2 mu M for 4 h) increased NFAT transcriptional activity by seven- and fourfold, respectively, in myotubes transiently transfected with an NFAT- dependent reporter plasmid ( pNFAT- luc, Stratagene). Cotreatment with the NOS- inhibitor N-G- nitro- L- arginine methyl ester ( L- NAME; 5 mM) or the guanylate cyclase inhibitor 1H[ 1,2,4] oxadiazolo[ 4,3- a] quinoxalin- 1- one ( ODQ; 10 mu M) prevented the calcium effects on NFAT activity. The NO donor diethylenetriamine-NONO ( DETA- NO; 10 mu M) augmented the effects of A23187 on NFAT- dependent transcription. Similarly, A23187 ( 0.4 mu M for 4 h) caused nuclear accumulation of NFAT and increased phosphorylation ( i. e., inactivation) of GSK- 3 beta, whereas cotreatment with L- NAME or ODQ inhibited these responses. Finally, the NO donor 3-( 2- hydroxy- 2- nitroso- 1- propylhydrazino)- 1- propanamine ( PAPA-NO; 1 mu M for 1 h) increased phosphorylation of GSK- 3 beta in a manner dependent on guanylate cyclase activity. We conclude that NOS activity mediates calcium- induced phosphorylation of GSK- 3 beta and activation of NFAT- dependent transcription in myotubes. Furthermore, these effects of NO are guanylate cyclase- dependent.