大鼠“承山”穴的神经解剖学特征——霍乱毒素亚单位B结合荧光素488法  

Neuroanatomical Characteristics of Acupoint "Chengshan" (BL 57) in the Rat: A Cholera Toxin Subunit B Conjugated with Alexa Fluor 488 Method Study

作　　者：朱新龙[1] 白万柱[2] 吴富东[1] 蒋瑾[2] 景向红[2] 

机构地区：[1]山东中医药大学,济南250021 [2]中国中医科学院针灸研究所,北京100700

出　　处：《针刺研究》2010年第6期433-437,共5页Acupuncture Research

基　　金：中国中医科学院自主选题研究项目(No.2009Z02024);国家人力资源和社会保障部回国留学人员科技项目(No.2009);国家重点基础研究发展计划("973"计划)资助项目(No.2010CB530507)

摘　　要：目的：应用霍乱毒素亚单位B结合荧光素488（CTB—Alexa 488）新一代荧光示踪技术研究大鼠“承山”穴神经解剖学特征。方法：用微量注射器将5μL0．05％CTB—Alexa 488分别注入4只雄性SD大鼠“承山”穴。在40～48h后将动物灌流固定，取出脑、脊髓和双侧腰部脊神经节，并制成组织切片，后直接用荧光显微镜系统观察和记录被CTB-Alexa 488显示的神经标记。结果：被CTB-Alexa 488标记的神经元在荧光光栅450～490的条件下呈绿色，均位于示踪剂注入侧的相关神经组织中。其中标记的一级感觉神经元分布在L4-L5的脊神经节，而被标记的一级运动神经元呈柱状分布在L4-L5节段脊髓前角第Ⅸ层的中外侧部。结论：应用CTB-Alexa 488荧光示踪技术成功揭示了与“承山”穴相关的初级感觉和运动神经元在脊神经节和脊髓的分布规律。Objective To investigate neuroanatomical characteristics of the primary sensory afferent and the motor neurons coming from and innervating acupoint ＂Ohengshan＂ （BL 57） area in the rat by using cholera toxin subunit B conjugated with Alexa Fluor 488 （CYB-Alexa 488）, a new generation of fluorescent neural tracing reagent. Methods Four male SD rats were used in the present study. Under anesthesia, 0.05% CTB-Alexa 488 （5 μL） was injected into the central part of the rear of the hind leg, a corresponding site of ＂Ohengshan＂ （BL 57） in the human body. After 40-48 surviving hours, the rat＇s brain, spinal cord and dorsal root ganglia （DRGs） of the lumbar segments （L1- L6 ） were dissected following perfusion with 4% paraformaldehyde, cut into sections and observed under fluorescent microscope equipped with a digital camera. The neurons labeled by CTB-Alexa 488 were counted. Results All OTB-Alexa 488 labeled neurons appeared in green under fluorescent filters of 450 490 and were located ipsilaterally on the injection side. The labeled primary sensory neurons were found in the DRGs at L4 （ 1 1 neurons） and L5 （35 neurons）. Among them, 29 neurons （63.04%） were bigger, with their cell body diameters being 35-50 μm and 17 （36.96%） smaller, with their body diameters being lower than 35 pro. The labeled motor neurons were found to distri bute in the mediolateral portion of lamina Ⅸ, forming a longitudinal column from L4 to L5. Of the observed 316 motor neurons, 259 （81.96%） belong to a type with their body diameters being 25-40 μm and 57 （ 18.04% ） to γ type with their body diameters being lower than 25 lain. Conclusion The OTB-Alexa 488-labeled primary sensory and motor neurons innervating acupoint ＂Chengshan＂ （BL 57） distribute in the DRGs of L4 - L5 ; The present fluorescent tracing technique may be quite useful for investigating the neural characteristics of acupoints.

关 键 词：承山 神经支配 霍乱毒素亚单位B结合荧光素488 脊神经节 脊髓 

分 类 号：R224.2[医药卫生—中医基础理论]