高渗条件下大鼠下丘脑神经元和星形胶质细胞内钙的动态变化

背景既往有很多关于培养神经元或胶质细胞内钙的研究.然而关于高渗条件下共同观察神经元和胶质细胞内钙变化的研究目前较少.目的探讨高渗刺激后大鼠下丘脑神经元和星形胶质细胞(AST)内钙的变化.设计随机对照实验研究.单位一所大学医院的神经外科和一所大学神经科学研究所的实验室.材料实验在南方医科大学医院神经外科和解放军第四军医大学全军神经科学研究所完成.新生两三天的SD大鼠2只和孕18 d大鼠胚胎6只,清洁级,由解放军第四军医大学实验动物中心提供.方法体外联合培养的神经元和AST经钙荧光指示剂Fluo-3负载后,用激光扫描共聚焦显微镜测定高渗NaCl溶液影响前后的细胞内钙水平随时间变化情况.主要观察指标联合培养的神经元和AST的形态观察及等渗和高渗状态下神经元和AST内Ca2+含量测定.结果高渗刺激使培养神经元和AST内钙水平先升高后降低,最后维持在比高渗刺激前稍高的静息钙水平上.AST钙水平达到峰值的速度略快于神经元.结论神经元和AST内Ca2+在渗透压信息传递过程起作用.

Dynamic changes of calcium in rat hypothalamic neurons and astrocytes following hyperosmotic stimulation

BACKGROUND: There have been lots of reports about study on intracellular Ca2 + in cultured neurons or astrocytes. However, little is known about the changes of intracellular Ca2 + in co-cultured astrocytes and neurons following hyperosmotic stimulation.OBJECTIVE: To investigate the changes of intracellular calcium in rat hypothalamic neurons and astrocytes following hyperosmotic stimulation DESIGN: A randomized controlled study.SETTING: Department of Neurosurgery, Laboratory of Neuroscience Research Institute of a hospital of a university.MATERIALS: This experiment was carried out at the Department of Neurosurgery of Zhujiang Hospital of Sourthern Medical University, and the Institute of Neuroscience of the Fourth Military Medical University of Chinese PLA. Two postnatal (2 -3 days) and six embryonic(E18) healthy male SD rats of cleaning grade were purchased from the Experimental Animal Center of the Fourth Military Medical University of Chinese PLA.METHODS: Time-dependant changes of intracellular calcium in co-cultured neurons and astrocytes marked by fluorescent Ca2 + indicator Fluo-3 following hyperosmotic NaCl stimulation were observed with a laser scanning eonfocal microscope.MAIN OUTCOME MEASURES: The morphology of co-cultured neurons and astrocytes, as well as the content of intracellular Ca2 + under isotonic and hyperosmotic conditions.RESULTS: The level of intracellular Ca2+ of co-cultured hypothalamic neurons and astrocytes was founid increased due to hyperosmotic stimulation,which later decreased and finally stayed at a level slightly higher than the resting Ca2 + level before stimulation. Peak time of intracellular Ca2 + in astrocytes was slightly earlier than that of neurons.CONCLUSION: The results indicated that Ca2 + in neurons and astrocytes might play a certain role during the transmission of the message of osmotic pressure.