Interaction between neuronal nitric oxide synthase signaling and temperature influences sarcoplasmic reticulum calcium leak: role of nitroso-redox balance.
Interaction between neuronal nitric oxide synthase signaling and temperature influences sarcoplasmic reticulum calcium leak: role of nitroso-redox balance.
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DOI：
10.1161/circresaha.116.305172
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发表时间：
2015-01-02
影响因子：
20.1
通讯作者：
Hare JM
中科院分区：
文献类型：
--
作者：
Dulce RA;Mayo V;Rangel EB;Balkan W;Hare JM
尽管一氧化氮(NO)信号调节心脏功能和兴奋-收缩偶联，但由于不一致的实验条件，特别是温度，导致相反的结果，混淆了阐明NO信号通路的能力。在这里，我们证明了温度对NO的影响是显著的。测试温度深刻影响亚硝基-氧化还原平衡，从而影响肌网(SR)钙离子泄漏的假设。我们测定了野生型(WT)、NO/氧化还原失衡(NOS1−/−)和高S亚硝酸化(GSNOR−/−)小鼠心肌细胞的肌浆网钙离子泄漏。在WT心肌细胞中，当温度从37℃降至23℃时，肌浆网钙离子泄漏增加，而在NOS1−/−细胞中，当温度超过30℃时，其泄漏突然增加。GSNOR−/−心肌细胞在整个温度范围内表现为低泄漏。外源性添加NO对NOS1−/−心肌细胞有双相效应；37℃时减少漏出，亚生理温度时增加漏出。奥比嘌醇和坦泊尔可减少NOS1−/−心肌细胞的渗漏。从37℃到23℃的降温增加了WT细胞的ROS生成，但减少了NOS1−/−心肌细胞的ROS生成。奥比嘌醇进一步减少了ROS的产生。在23℃的WT细胞中，四氢生物蝶呤(一种重要的一氧化氮合酶辅助因子)可减少渗漏。降温可显著增加NOS1−/−细胞的肌浆网钙离子含量，但对WT和GSNOR−/−细胞无明显影响。Ca2+渗漏和温度通常成反比，而NOS1缺乏则相反，随着温度的升高，渗漏增加，ROS产生增加。减少脱硝化(GSNOR不足)消除了泄漏对温度的依赖。因此，温度调节NO和ROS之间的平衡，进而对肌质网钙离子产生重大影响。
While nitric oxide (NO) signaling modulates cardiac function and excitation-contraction coupling, opposing results due to inconsistent experimental conditions, particularly with respect to temperature, confound the ability to elucidate NO signaling pathways. Here we show that temperature significantly modulates NO effects. Test the hypothesis that temperature profoundly impacts nitroso-redox equilibrium, thereby affecting sarcomeric reticulum (SR) Ca2+ leak. We measured SR Ca2+ leak in cardiomyocytes from wild-type (WT), NO/redox imbalance (NOS1−/−), and hyper S-nitrosylation (GSNOR−/−) mice. In WT cardiomyocytes, SR Ca2+ leak increased as temperature decreased from 37°C to 23°C, whereas, in NOS1−/ −cells, the leak suddenly increased when the temperature surpassed 30°C. GSNOR−/ − cardiomyocytes exhibited low leak throughout the temperature range. Exogenously added NO had a biphasic effect on NOS1−/− cardiomyocytes; reducing leak at 37°C but increasing it at sub-physiologic temperatures. Oxypurinol and Tempol diminished the leak in NOS1−/ − cardiomyocytes. Cooling from 37° to 23°C increased ROS generation in WT but decreased it in NOS1−/− cardiomyocytes. Oxypurinol further reduced ROS generation. At 23°C in WT cells, leak was decreased by tetrahydrobiopterin, an essential NOS cofactor. Cooling significantly increased SR Ca2+ content in NOS1−/− cells but had no effect in WT or GSNOR−/−. Ca2+ leak and temperature are normally inversely proportional, whereas NOS1 deficiency reverses this effect, increasing leak and elevating ROS production as temperature increases. Reduced denitrosylation (GSNOR deficiency) eliminates the temperature dependence of leak. Thus, temperature regulates the balance between NO and ROS which in turn has a major impact on SR Ca2+.