5-HT受体亚型在原代培养大鼠脊髓背角神经元的表达  

EXPRESSION OF 5-HT RECEPTOR SUBTYPE mRNAs IN PRIMARY CULTURED SPINAL DORSAL HORN NEURONS OF THE RAT

作　　者：王亚云[1] 武胜昔[1] 刘翔宇[1] 王文[1] 李云庆[1] 

机构地区：[1]第四军医大学基础医学部解剖学教研室,梁銶琚脑研究中心西安710032

出　　处：《神经解剖学杂志》2002年第3期191-196,T001,共7页Chinese Journal of Neuroanatomy

基　　金：国家自然科学基金 (No.3 980 0 0 3 4) ;高等学校骨干教师资助计划 ;国家重点基础研究规划"脑功能和脑重大疾病的基础研究"(G19990 5 40 0 4)资助项目

摘　　要：为探讨脊髓背角内 5 -HT发挥作用的受体类型及其胞内信号转导机制 ,本研究首先利用免疫荧光技术对胎鼠脊髓背角神经元的产率进行了观察 ,再利用反转录 PCR方法观察了 5 -HT受体 14种亚型 m RNAs在原代培养神经元中的表达。原代培养的背角神经元生长状态良好 ,存活时间达到 14～ 2 1d。对培养的背角细胞用神经元特异性标志物—神经细胞核蛋白 (Neu N)进行免疫荧光检测的结果表明 ,神经元的产率超过 90 % ;用 PCR方法在培养的背角神经元中检测到了 5 -HT1 A、5 -HT1 B、5 -HT1 D、5 -HT1 F、5 -HT2 A、5 -HT2 C、5 -HT3、5 -HT4 、5 -HT5A、5 -HT5B、5 -HT6 和 5 -HT7等受体亚型 m RNAs的表达。但上述各受体亚型的表达水平存在差异 ,未检测到 5 -HT1 E和 5 -HT2 B受体亚型 m RNAs的表达。结果表明 ,本研究建立的实验方法可满意地获得原代培养脊髓背角神经元 ;这些神经元不但表达多种受体亚型 ,而且表达类型与以往在成年大鼠脊髓背角观察到的表达状况基本一致。上述结果为进一步开展 5 -HT作用机制的体外研究奠定了基础。To investigate the receptor and intracellular signal transduction mechanism of 5 HT in the spinal dorsal horn, the spinal dorsal horn neurons of the embryonic rat were primary cultured and their classification was determined by immunofluorecent staining method. The expression of 14 subtypes of 5 HT receptor mRNAs in cultured dorsal horn neurons was also examined by using reverse transcription polymerase chain reaction (RT PCR). The results showed that primary cultured spinal dorsal horn neurons grew normally and exhibited satisfied morphological features. Immunofluorescent staining with neuronal nucleus protein (NeuN), a neuronal marker, revealed that more than 90% of the cultured dorsal horn cells were neurons. RT PCR results detected the expression of 5 HT 1A , 5 HT 1B , 5 HT 1D , 5 HT 1F , 5 HT 2A , 5 HT 2C , 5 HT 3, 5 HT 4, 5 HT 5A , 5 HT 5B , 5 HT 6 and 5 HT 7 receptor subtype mRNAs in cultured spinal dorsal horn neurons. But the difference of expression level existed between different 5 HT receptor subtypes. The expression of 5 HT 1E and 5 HT 2B receptor mRNAs, however, was not detected in the present study. The present results indicate that the experimental methods used in the present study were useful to obtain satisfied primary cultured spinal dorsal horn neurons. These neurons express mRNAs for various 5 HT receptor subtypes and provide valuable evidence for the further research on 5 HT in the spinal dorsal horn. (Figure 1 on plate 43)

关 键 词：5-HT受体亚型 原代培养 大鼠脊髓背角神经元 表达 

分 类 号：R322[医药卫生—人体解剖和组织胚胎学]