Ferreyra G A, Cammarota M P, Golombek D A
Departamento de Fisiología, Facultad de Medicina, Universidad de Buenos Aires, Argentina.
Brain Res. 1998 Jun 29;797(2):190-6. doi: 10.1016/s0006-8993(98)00376-x.
Circadian rhythms are controlled by an endogenous clock, which in mammals is located in the hypothalamic suprachiasmatic nuclei (SCN). A role for nitric oxide in circadian responses to light has been indicated. To test the role of nitric oxide synthase (NOS) in the SCN and in circadian responses to light, we examined NOS specific activity at different time points and photic conditions. NOS activity was determined by the conversion of 3H-arginine to 3H-citrulline. NOS enzymatic activity in the SCN was significantly higher during the dark phase than during the day, without any changes in the levels of the NOS protein. However, this difference disappeared when animals were placed under constant darkness, and NOS activity was similar at CT 8 and CT 18 (with CT 12 defined as the onset of the subjective night). When 5-min light pulses were administered at these time points (when light would induce no phase shift or a phase advance, respectively), NOS activity was significantly increased almost equally. A spectrophotometric assay was used to determine NO content in the SCN, showing relatively high constitutive levels enhanced by 100 microM glutamate. These results suggest that NOS activity is not controlled by the circadian clock, although it might mediate some of the effects of light on biological rhythms.
昼夜节律受内源性生物钟控制,在哺乳动物中,该生物钟位于下丘脑视交叉上核(SCN)。已有研究表明一氧化氮在对光的昼夜反应中发挥作用。为了测试一氧化氮合酶(NOS)在SCN以及对光的昼夜反应中的作用,我们检测了不同时间点和光照条件下的NOS比活性。NOS活性通过3H-精氨酸向3H-瓜氨酸的转化来测定。SCN中的NOS酶活性在黑暗期显著高于白天,而NOS蛋白水平没有任何变化。然而,当动物处于持续黑暗环境中时,这种差异消失了,在CT 8和CT 18时NOS活性相似(将CT 12定义为主观夜晚的开始)。当在这些时间点给予5分钟的光脉冲时(分别是光不会引起相位偏移或相位提前的情况),NOS活性几乎同等程度地显著增加。使用分光光度法测定SCN中的NO含量,结果显示相对较高的组成水平会被100 microM谷氨酸增强。这些结果表明,NOS活性不受昼夜节律生物钟的控制,尽管它可能介导了光对生物节律的一些影响。