Tyrosine phosphorylation of neuronal nitric oxide synthase (nNOS) during hypoxia in the cerebral cortex of newborn piglets: the role of nitric oxide.
Tyrosine phosphorylation of neuronal nitric oxide synthase (nNOS) during hypoxia in the cerebral cortex of newborn piglets: the role of nitric oxide.
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DOI：
10.1016/j.neulet.2009.06.075
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发表时间：
2009-10-02
影响因子：
2.5
通讯作者：
Delivoria-Papadopoulos M
中科院分区：
文献类型：
--
作者：
Mishra OP;Ashraf QM;Delivoria-Papadopoulos M
本研究旨在探讨缺氧时nNOS激活的机制，并验证缺氧诱导的新生仔猪大脑皮质膜nNOS酪氨酸磷酸化水平升高是由nNOS衍生的一氧化氮（NO）介导的假说。15头新生仔猪分为常氧组（Nx，n=5）、低氧组（Hx，n=5）和低氧预处理组（Hx-nNOSi）。通过0.07的FiO 2持续60分钟诱导缺氧。施用nNOS抑制剂I（选择性>2500 vs内皮NOS和>500 vs诱导型NOS）（0.4 mg/kg，i. v.）的缺氧前30分钟。分离皮质膜，通过Western印迹法测定nNOS的酪氨酸磷酸化。膜蛋白用nNOS抗体免疫沉淀，在12% SDS-PAGE上分离，并用抗磷酸酪氨酸抗体印迹。用增强化学发光法检测蛋白条带，用光密度法分析，以吸光度（OD× mm 2）表示。密度（O.D. Hx组为118.39 ± 14.17（p< 0.05 vs Nx），Hx-nNOSi组为45.56±10.34（p <0.05 vs Hx，p=NS vs Nx）。结果表明，用nNOS抑制剂预处理防止缺氧诱导的nNOS酪氨酸磷酸化增加。我们的结论是缺氧诱导的nNOS酪氨酸磷酸化增加的机制是由nNOS衍生的NO介导的。
The present study aims to investigate the mechanism of activation of nNOS during hypoxia and tests the hypothesis that the hypoxia-induced increased tyrosine phosphorylation of nNOS in the cerebral cortical membranes of newborn piglets is mediated by nNOS-derived nitric oxide (NO). Fifteen newborn piglets were divided into normoxic (Nx, n=5), hypoxic (Hx, n=5) and hypoxic-pretreated with nNOS inhibitor I (Hx-nNOSi) groups. Hypoxia was induced by an FiO2 of 0.07 for 60 min. nNOS inhibitor I (selectivity >2500 vs endothelial NOS and >500 vs inducible NOS) was administered (0.4 mg/kg, i. v.) 30 min prior to hypoxia. Cortical membranes were isolated and tyrosine phosphorylation of nNOS determined by Western blot. Membrane protein was immunoprecipitated with nNOS antibody, separated on 12% SDS-PAGE and blotted with anti-phosphotyrosine antibody. Protein bands were detected by enhanced chemiluminescence, analyzed by densitometry and expressed as absorbance (OD×mm2). Density (O.D. x mm2) of tyrosine phosphorylated nNOS was 51.66±14.11 in Nx, 118.39 ±14.17 in Hx (p< 0.05 vs Nx) and 45.56±10.34 in Hx-nNOSi (p< 0.05 vs Hx, p=NS vs Nx). The results demonstrate that pretreatment with nNOS inhibitor prevents the hypoxia-induced increased tyrosine phosphorylation of nNOS. We conclude that the mechanism of hypoxia-induced increased tyrosine phosphorylation of nNOS is mediated by nNOS derived NO.
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影响因子：
2.5
作者：
HOFFMAN, DJ;MCGOWAN, JE;DELIVORIAPAPADOPOULOS, M
通讯作者：
DELIVORIAPAPADOPOULOS, M
影响因子：
3.3
作者：
Mishra, OP;Ashraf, QM;Delivoria-Papadopoulos, M
通讯作者：
Delivoria-Papadopoulos, M
影响因子：
4.4
作者：
MISHRA, OP;DELIVORIAPAPADOPOULOS, M
通讯作者：
DELIVORIAPAPADOPOULOS, M
DOI：
10.1038/6675
发表时间：
1999-03-01
期刊：
NATURE STRUCTURAL BIOLOGY
影响因子：
--
作者：
Fischmann, TO;Hruza, A;Weber, PC
通讯作者：
Weber, PC
影响因子：
3.9
作者：
ISCHIROPOULOS, H;ZHU, L;BECKMAN, JS
通讯作者：
BECKMAN, JS