摘要
用荧光素逆行双标和免疫组化结合三标技术,研究脊神经节含CCK-8的感觉神经元的皮肤-内脏分支投射。将fast blue(FB)和nuclear yellow(NY)分別注入大鼠左侧腹腔神经节和左侧第9～11肋间神经皮支内。灌注固定,取左侧胸9～11脊神经节,恒冷箱切片。在Nikon荧光显微镜下观察,激发光波长为365nm。镜下可见三种不同标记的神经元:NY标记神经元,胞核发黄色荧光;FB标记神经元,胞浆发蓝色荧光;NY+FB双标神经元(具有分支投射的神经元),胞浆发蓝色荧光和胞核发黄色荧光。双标神经元约占所有荧光标记神经元的2.8％。含有双标神经元的切片,用CCK-8抗血清孵育,用改良的PAP法显色。镜下可见CCK免疫反应阳性神经元。在CCK免疫反应阳性神经元中,可以区分:1.被CCK标记的NY神经元(NY+CCK)。2.被CCK标记的FB神经元(FB+CCK)。3.被CCK标记的NY+FB双标神经元(NY+FB+CCK)。4.单纯CCK免疫反应阳性神经元。NY+FB+CCK三标神经元约占NY+FB双标神经元的11.5％,仅占CCK阳性神经元的0.4％。以上结果表明:脊神经节中有些感觉神经元的周围突具有分支,分别投射到内脏和皮肤;这类神经元中,有些神经元含有CCK-8。从而提示:内脏牵涉性痛的会聚可在脊神经节进行;CCK可能参与内脏和皮肤感觉会聚的调节。因此,本文为牵涉性痛的发生机制和躯体-内脏反射提供了重要的化学神经解剖学资料。
Skin-visceral divergent projections of cholecystokinin(CCK)-containing dorsal root ganglion neurons were studied by combined technique of fluorescent double-labelling and immunohistochemistry. Fast Blue(FB) and Nuclear Yellow (NY) were injected into the coeliac ganglion and the cutaneous branches of left 9th-11th intercostal nerves, respectively. Three kinds of neurons labelled with fluorescein were observed in T_(9-11) dorsal root ganglia under Nikon fluorescence microscope with 365 nm excitation light: FB-labelled neurons with blue-fluorescing cytoplasm; NY-labelled neurons with yellow-fluorescing nucleus and double-labelled neurons with blue cytoplasm and yellow nucleus. The double-labelled neurons were found to be 2.8％ of total labelled neurons.
The sections containing fluorescein labelled neurons were then stained by CCKimmunohistochemical procedure. Four kinds of neurons could be identified: NY-neurons, with CCK-immunoreactivity (NY+CCK); FB-neurons with CCK-immunoreactivity(FB + CCK);NY + FB neurons with CCK-immunoreactivity(NY + FB + CCK); Single CCK-positive neurons. NY + FB + CCK triple-labelled neurons accounted for approximately 11.5％ of NY + FB double-labelled neurons,and 0.4％ of all CCK-positive neurons.
The findings clearly indicated that the peripheral processes of some sensory dorsal root ganglion neurons project divergently to both skin and visceral structure, and contain CCK. The present results suggest that the peripheral dichotomization of the dorsal root ganglion nearons might converge sensory inputs from both skin and visceral fields, and thus not only provide one of the structural basis for the referred pain but also reveal that CCK might play a mediation role in the skin-visceral reflection and referred pain.
出处
《解剖学报》
CAS
CSCD
北大核心
1990年第1期25-30,共6页
Acta Anatomica Sinica
基金
中国科学院科学基金
关键词
荧光素
免疫组化
脊神经节细胞
Fluorescent tracers
Immunohistochemistry
Cholecystokinin
Dorsal root ganglion neurons
Divergent projections
