小鼠在高架十字迷宫中下丘脑CRFmRNA表达的变化.docVIP

5-羟色胺转运体敲除小鼠高架十字迷宫CRF mRNA表达的变化 文玲1，邓永兵1,2，朱小云1，蒋 雪1（400016重庆,重庆医科大学生物化学与分子生物学教研室1；400014重庆，重庆市急救医疗中心神经外科2） [摘要] 目的 研究5-羟色胺转运体敲除小鼠高架十字迷宫变化。方法 将小鼠按基因型（SERT+/+、 +/-、 -/-）分为对照组和EPM）组，每组只。利用高架十字迷宫作为应激原刺激小鼠，各组小鼠室旁核。结果在SERT敲除小鼠的EPM刺激下，SERT+/-和-/-小鼠的显著高于SERT+/+ P 0.05 结论 应激显。 [关键词] 5-羟色胺转运体；高架十字迷宫室旁核The Alteration of CRF mRNA Expression in Hypothalamus of SERT Knockout Mice to EPM [Abstract] Objective The alteration of CRF mRNA expression in hypothalamus of SERT knockout mice to EPM. Methods The mice were divided into two groups, according their genotype SERT+/+, SERT+/-,SERT-/- and each group has ten mice. Using the elevated plus maze as a stressor to stimulate mice. Then anxious like behavior was observed, and the CRF mRNA on the hypothalamus and nucleus paraventricularis PVN were measured. Results The basal expression of CRF mRNA in hypothalamus and PVN were lower in both SERT+/- and SERT-/-mice than that in SERT+/+ mice, compared with SERT+/+ mice, stress from exposure to the elevated plus maze siginificantly increased CFRmRNA in the hypothalamus and nucleus paraventricularis of SERT+/- and -/- mice P 0.05 。Conclusion Stress from exposure to the EPM significantly increased CRF mRNA in the PVN and hypothalamus of SERT+/- and SERT-/- mice. the abnormal stress of SERT knockout mice associated with the increased CRFmRNA expression. [Key Words] SERT；EPM；Hypothalamus；PVN；CRF mRNA 下丘脑-垂体-肾上腺皮质（hypothalamic-pituitary-adrenal cortex, HPA ）轴是重要的神经内分泌循环通路，其主要通过分泌皮质高架十字迷宫（elevated plus maze，EPM）本实验SERT敲除小鼠室旁核（PVN）为研究敏感性增加提供重要的。1 材料与方法 1.1 动物 基因型为SERT+/-小鼠购自Taconic farm公司，雌雄小鼠进行交配，产下子代小鼠的基因型通过基因型PCR进行鉴定。用于实验的小鼠为25个月大、体为2025g。 1.2 仪器 高架十字迷宫：San Diego ；Bio-Rad公司；微量离心机：Bio-Rad公司；冰箱、凝胶电泳仪、恒温水浴箱等常用仪器：重庆医科大学1.3 试剂 35S-UTP：Amersham公司；胶回收试剂盒：Q-Bio gene公司；体外转录试剂盒：Fermentas公司；7500 fast Real-Time PCR system试剂盒：Applied Biosystems公司；ProbQuant G-50 micro column：Pharmacia公司；大肠杆菌E. Coli DH10B. 、PCR II-TOPO TA克隆载体以及引物合成和PCR试剂盒：Invetrogen公司；分子生物学工具酶：NEB公司、Fermentas公司；常规试剂：Sigma公司。 1.4 方法1.4.