霍乱毒素B亚单位结合胶体金的逆行轴浆转运复合免疫细胞化学的应用——一种新的非荧光双标法

RETROGRADELY AXOPLASMIC TRANSPORT OF CHOLERA TOXIN SUBUNIT B(CB) COUPLED WITH COLLOIDAL GOLD IN COMBINATION WTIH IMMUNOCYTOCHEMISTRY-A NEW NONFLUORESCENT DOUBLE-LABELLED METHOD

以CB-胶体金(Cholera toxin subunit B-colloidal gold)为逆行追踪剂(CB-胶体金,中国协和医科大学神经生物学室制备),观察将其肌肉注射后,大鼠运动神经元的逆行轴浆转运标记及复合免疫细胞化学(ICC.immunocytochemistry)应用的可行性.结果为:(1)运动神经元末梢能摄取CB-胶体金,并经轴浆转运到胞体,经银加强后于光镜下可见清晰标记的神经元胞体和近端树突;(2)转运后的胶体金,可长期稳定地标记运动神经元长达30天;(3)经银加强复合免疫细胞化学的双标记运动神经元,由于两种标记反差大极易辨认.实验结果显示了这一新的非荧光双标记技术的优越性.

The purpose of the present paper was to test whether CB colloidal gold (CB-gold), prepared by our laboratory, could be retrogradely transported in motoneurons after it was injected into muscles and whether it would be combined with immunocytochemistry(ICC). CB-gold was visualized by silver intensification under light microscope. The results were as follows: (1) Retrogradely transported CB-gold appeared as black deposits in the perilarya and proximal dendrites, (2)CB-gold persisted in motoneurons for at least 30 days. (3) The double-labelled neurons were easily recoganized by CB-gold retrograde tracing in combination with ICC. The above results show that CB-gold tracing combined with ICC may become a valuable tool in the neuroanatomical and ICC research.