细胞内钙离子释放在嗅觉信号转导机制中的作用  

The role of intracellular Ca^(2+) release in olfactory signal transduction

作　　者：羡慕[1] 韩德民[1,2] 张罗[1,2] 

机构地区：[1]首都医科大学附属北京同仁医院耳鼻咽喉头颈外科,北京100730 [2]教育部耳鼻咽喉头颈科学重点实验室北京市耳鼻咽喉科研究所

出　　处：《临床耳鼻咽喉头颈外科杂志》2010年第20期940-944,共5页Journal of Clinical Otorhinolaryngology Head And Neck Surgery

基　　金：国家自然科学基金资助项目(No:30672302)

摘　　要：目的:建立组织块法体外培养嗅觉受体神经元(ORNs)细胞内游离钙离子浓度([Ca2+]i)的实时观察系统,分析嗅觉信号转导体系中几个关键环节的作用。方法:以双激发波长钙离子荧光探针Fura-2 AM孵育培养的ORNs,通过双波长比率法,实时定量观察ORNs的[Ca2+]i。分别以Forskolin和IBMX刺激ORNs,观察[Ca2+]i的改变。分别耗竭细胞内钙库和使用不含钙离子的细胞外液,判断细胞内[Ca2+]i的来源。结果:静息ORNs的[Ca2+]i为(58.5±12.8)nmol/L。Forskolin和IBMX刺激可使ORNs的[Ca2+]i快速升高,且作用可逆。二者所引发的钙信号来自于细胞外钙内流,而非细胞内钙库的释放。结论:本研究成功建立了实时定量观察ORNs细胞内[Ca2+]i的系统,ORNs的[Ca2+]i受第二信使门控钙离子通道的调节。Objective：To setup the real time monitor system of the concentration of free intracellular calcium（i） of olfactory receptor neurons（ORNs） cultured from olfactory epithelium explant,and to analyze the role of several important components in olfactory signal transduction.Method：The i of the cultured ORNs was determined by fluorescence microscopy using the fluorescent calcium indicator,Fura-2 AM,and calculated by means of dual-wavelength ratiometric method.Forskolin and IBMX were used to stimulate the cultured ORNs respectively.The source of corresponding i elevation was studied by the depletion of extracelluar or intracellular calcium.Result：The i of silent ORNs was（58.5±12.8） nmol/L.Forskolin or IBMX stimulation led to reversible accumulation of i in the ORNs.The i change was abolished with the removal of extracellular Ca2＋ and un-affected by treatment with thapsigargin.Conclusion：A system to visualize and quantify i of the ORNs was established.i of the ORNs was regulated by second messenger gated calcium channels.

关 键 词：嗅觉受体神经元 信号转导 钙离子 钙离子荧光探针 

分 类 号：R765[医药卫生—耳鼻咽喉科]