中缝背核触液神经元对荧光活性染料DiI亲和性的实验研究  

Labeling cerebrospinal fluid-contacting neurons in the dorsal raphe nucleus with the fluorescent dye DiI

作　　者：刘苏[1,2,3] 高甜惠[2,3] 范建伟[2,3] 刘岳鹏[2,3] 

机构地区：[1]徐州医学院附属医院麻醉科,江苏徐州221002 [2]徐州医学院江苏省麻醉学重点实验室,江苏徐州221004 [3]江苏省麻醉与镇痛应用技术重点实验室,江苏徐州221004

出　　处：《徐州医学院学报》2013年第12期799-801,共3页Acta Academiae Medicinae Xuzhou

基　　金：国家自然科学基金青年基金（8100047）

摘　　要：目的 观察荧光染料DiI标记中缝背核触液神经元的可行性.方法 6只雄性SD大鼠,侧脑室注射1％3μl的DiI染料,3天后经4％多聚甲醛灌注固定,冰冻切片机切片,共聚焦显微镜下观察,并进行连续光学切片兼对标记的触液神经元进行三维重塑.结果 DiI对中脑导水管腹侧处中缝背核内的触液神经元标记清楚,且阳性标记神经元的三维重塑显示,中缝背核触液神经元的胞体和近胞体的一级树突可被清楚标记.结论 中缝背核触液神经元对荧光活性染料DiI的亲和力较强,脑切片无需进行其他染色处理,可直接在荧光显微镜下观察到触液神经元,是一种示踪标记中缝背核触液神经元的良好方法.Objective To observe the feasibility of labeling cerebrospinal fluid （CSF） - contacting neurons in the dorsal rapbe nucleus with the fluorescent dye DiI. Methods 6 SD male rats were selected to receive intra - cerebroventricular injection of 1% 3 1 DiI dye. Then the brains were removed and fixed in 4% paraformaldehyde for 3 days followed by sectioning using a frozen slicer. The resultant slices were observed under a confoeal microscope. Meanwhile the labeled CSF - contacting neurons underwent optical sectioning for three dimensional remodeling. Results The DiI stain in the CSF - contacting neurons of the dorsal raphe nucleus around cerebral aqueduct was clear. Based on the three - dimensional remodeling of DiI - labeled neurons, the neurons and their primary dendrites could be labeled. Conclusion DiI dye has strong affinity to CSF -contacting neurons in the dorsal rape nuclear. The neurons on the brain sections can be seen under a fluorescent microscope without other dying procedures. Therefore, DiI staining can be adopted to label CSF - contacting neurons.

关 键 词：DiI染料 触液神经元 中缝背核 大鼠 

分 类 号：R338[医药卫生—人体生理学]