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Objective To examine the effects of melatonin on the dynamic changes in the concentration of intracellular free Ca 2+ ([Ca 2+ ]i) in single intact cultured cortical neurons isolated from fetal rats, in order to explore the possible antiaging mechanisms of melatonin (MT) Methods 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?μmol/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 [Ca 2+ ]i while not disturbing normal intracellular physiology The changes in fluorescent intensity were monitored by LSCM Results Bay K8644 (10 6 ?mol/L), KCl (20 ?mmol/L), sodium L glutamate (Glu, 50?μmol/L) caused a rapid increase of [Ca 2+ ]i in cortical neurons, and this increase could be significantly attenuated by 10 6 and 10 7 mol/L MT Conclusions MT could antagonize the extracellular Ca 2+ influx, reduce Ca 2+ overload, and have a protective effect on neurons This may be one of the important antiaging mechanisms of MT
Objective To examine the effects of melatonin on the dynamic changes in concentration of intracellular free Ca 2+ ([Ca 2+] i) in single intact cultured cortical neurons isolated from fetal rats, in order to explore the possible antiaging mechanisms of melatonin ( MT) Methods Using the highly fluorescent Ca 2+ sensitive indicator Fluo 3 / AM, cortical neurons cultured in a 35 μm Tissue Culture Dish were incubated for 45 min at room temperature with 5 μmol / L Fluo 3 / AM, in proper intracellular dye concentration to provide adequate signal strength for detection and excellent Laser Scanning Confocal Microscopy (LSCM) imaging of [Ca 2+] i while not disturbing normal intracellular physiology The changes in fluorescence intensity was monitored by LSCM Results Bay K8644 (10 6 · mol / L), KCl (20 mmol / L), sodium L glutamate (Glu, 50 μmol / L) caused a rapid increase of [Ca 2+] i in cortical neurons, and this increase could be significant ly attenuated by 10 6 and 10 7 mol / L MT Conclusions MT could antagonize the extracellular Ca 2+ influx, reduce Ca 2+ overload, and have a protective effect on neurons This may be one of the important anti-aging mechanisms of MT