Zhang Q, Zhang J
Department of Pharmacology II, Institute of Materia Medica, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing 100050, China.
Chin Med J (Engl). 2000 Jun;113(6):558-62.
To examine the effects of melatonin on the dynamic changes in the concentration of intracellular free Ca2+ ([Ca2+]i) in single intact cultured cortical neurons isolated from fetal rats, in order to explore the possible antiaging mechanisms of melatonin (MT).
Using the highly fluorescent Ca(2+)-sensitive indicator Fluo-3/AM, cortical neurons cultured in a 35 mm Tissue Culture Dish were in incubated for 45 min at room temperature with 5 mumol/L Fluo-3/AM, resulting in proper intracellular dye concentration to provide adequate signal strength for detection and excellent Laser Scanning Confocal Microscopy (LSCM) imaging of [Ca2+]i while not disturbing normal intracellular physiology. The changes in fluorescent intensity were monitored by LSCM.
Bay K8644 (10(-6) mol/L), KCl (20 mmol/L), sodium L-glutamate (Glu, 50 mumol/L) caused a rapid increase of [Ca2+]i in cortical neurons, and this increase could be significantly attenuated by 10(-6) and 10(-7) mol/L MT.
MT could antagonize the extracellular Ca2+ influx, reduce Ca2+ overload, and have a protective effect on neurons. This may be one of the important antiaging mechanisms of MT.
研究褪黑素对原代培养的胎鼠皮层神经元细胞内游离钙离子浓度（[Ca2+]i）动态变化的影响，以探讨褪黑素（MT）可能的抗衰老机制。
采用高荧光钙敏指示剂Fluo-3/AM，将培养于35mm组织培养皿中的皮层神经元在室温下用5μmol/L Fluo-3/AM孵育45分钟，使细胞内染料浓度合适，以提供足够的信号强度用于检测，并在不干扰正常细胞内生理的情况下对[Ca2+]i进行出色的激光扫描共聚焦显微镜（LSCM）成像。通过LSCM监测荧光强度的变化。
Bay K8644（10^(-6)mol/L）、KCl（20mmol/L）、L-谷氨酸钠（Glu，50μmol/L）可使皮层神经元[Ca2+]i迅速升高，而10^(-6)和10^(-7)mol/L的MT可显著减弱这种升高。
MT可拮抗细胞外Ca2+内流，减少Ca2+超载，对神经元具有保护作用。这可能是MT重要的抗衰老机制之一。
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