视黄酸对海马神经元细胞内钙离子浓度的影响  

Effects of All-trans Retinoic Acid on Intracellular Calcium Concentration of Hippocampal Neurons

作　　者：张小娟[1] 李廷玉[1] 刘友学[1] 陈洁[1] 瞿平[1] 魏小平[1] 

机构地区：[1]重庆医科大学儿童医院儿童营养研究中心,重庆400014

出　　处：《中国细胞生物学学报》2010年第2期235-240,共6页Chinese Journal of Cell Biology

基　　金：国家自然科学基金资助项目(No;30600495)~~

摘　　要：探讨全反式视黄酸(all-trans retinoic acid,atRA)对原代培养的海马神经元胞内钙离子浓度的影响,以进一步了解atRA参与学习记忆可能机制。分离新生Wistar大鼠海马,采用添加B27的无血清培养液进行海马神经元原代培养,免疫荧光鉴定培养的神经细胞;以fura-2/AM温育海马神经元,采用钙离子测定系统动态观察视黄酸对海马细胞内钙离子浓度的影响。结果显示:(1)培养的神经元纯度达90%;(2)atRA作用于海马神经元,能引起海马神经元胞内钙离子浓度的升高;(3)这种升高与atRA浓度及神经元的发育时间相关;(4)钙离子升高的具体方式是通过细胞外钙离子内流;(5)视黄酸核受体alpha(RARα)的拮抗剂Ro41-5253(Ro)对atRA升高的神经元胞内钙离子浓度有抑制作用。atRA与RARα结合,促使海马神经元胞外钙离子的内流,这可能是atRA参与学习记忆的机制之一。To investigate the effects and possible mechanisms of all-trans retinoic acid （atRA） on the intracellular Ca^2＋ level of primary cultured hippocampal neurons. Hippocampal cells were isolated from newborn rats within 24 hours and cultured for 1-12 days prior to experiments. Immunofluorescence was performed to evaluate the purity of cultured cells.The cultured primary hippocampal neurons were labeled with fura-2/AM, and fluorescence images were monitored using a CCD camera, and subsequently analyzed using imaging analyzing software. Ca^2＋ activities were presented as Ratio 340/380, the ratio of fluorescence intensities excited by alternating illumination of 340 nm and 380 nm beams. We showed that atRA caused an increase in the intracellular calcium concentration in primary hippocampal neurons, which was related with concentrations of atRA and days of neurons in vitro （DIV）. Furthermore, the increase in calcium level caused by atRA was through influx of extracellular calcium. Finally, we found that Ro41-5253 （Ro） reduced the increased Ratio 340/380 of hippocampal neurons caused by atRA. This study suggests that atRA can modulate calcium influx in hippocampal neurons, which was related with concentrations of atRA and days of neurons in vitro. AtRA plays a role in modulating calcium levels in hippocampal neurons possibly by binding to RARe. Finally, the role of atRA in regulating calcium concentration might be related to its effect on long-term potentiation （LTP）, learning, and memory.

关 键 词：视黄酸 钙影像 海马神经元 FURA-2 

分 类 号：Q42[生物学—神经生物学]