在同一神经元内同时显示Fos－蛋白、酪氨酸羟化酶和HRP逆行标记的三重标记技术

本文介绍了一种能在同一神经元内同时显示ＨＲＰ逆行标记以及Ｆｏｓ和某类神经递质或相关酶（例如ＴＨ）的三重标记技术。首先将ＨＲＰ注入脑内某一核团（如中央杏仁核），动物存活４８ｈ后，再向胃内导入１％福尔马林２ｍｌ，以造成对胃的伤害性刺激，２ｈ后将动物处死。脑的切片（如本文的延髓切片）先用四甲基联苯胺－钨酸钠（ＴＭＢ－ＳＴ）法进行ＨＲＰ呈色反应，后用二氨基联苯胺（ＤＡＢ）和氯化钴作加强；然后按ＡＢＣ法进行Ｆｏｓ免疫组织化学反应，继之按ＰＡＰ法进行ＴＨ免疫组织化学反应。光镜下在延髓切片观察到７种标记神经元，即ＴＨ－、ＨＲＰ－或Ｆｏｓ－单标记神经元，ＴＨ和ＨＲＰ－、ＴＨ和Ｆｏｓ－或ＨＲＰ和Ｆｏｓ的双标记神经元以及ＴＨ和ＨＲＰ和Ｆｏｓ的三标记神经元。该方法为研究同时显示中枢神经元的功能活动状态、传出投射及所含神经递质提供了一个强有力的工具。

STUDIES ON METHODS FOR ELECTROFUSION OF MOUSE 2-CELL EMBRYOS

This paper recommends a new triple-labeling technique,which demonstrates simutaneously Fos protein,neurotransmitter and horseradish peroxidase(HRP)retrograde labeling in same neurons.The procedure of this technique proceeds as follows.At first HRP was stereotaxically injected into central amygdaloid nucleus unilaterally,48 hours later,2 ml of 1% formalin was introduced into the stomach to induce visceral noxious stimulation,after 2hours,the animals were sacrificed.The sections of medulla oblongata were processed for histochemical demonstration of HRP with tetramethylbenzidine-sodium tungstate(TMB-ST)method,then treated with diaminobenzidine(DAB)and CoCl2 to intensify the HRP reactive products.Then Fos immunohistochemical reaction were processed according to ABC method , subsequently tyrosine hydroxylase(TH)immunohistochemical reaction were processed with PAP method.It was found that seven kinds of labeled neurons,i.e.TH-,HRP-,Fos-single labeled,TH/HRP,TH/Fos,HRP/Fos- double labeled and TH/HRP/Fos-triple labeled neurons were detected in the medulla,This technique provides a powerful tool for investigation of functional activity,efferent projection and neurotransmitter contents of the neurons of the central nervous system.