HRP和AP呈现束缚-浸水应激大鼠NTS儿茶酚胺能神经元Fos表达的效果比较

用辣根过氧化物酶(HRP)单酶先后标记Fos和酪氨酸羟化酶(TH),用二氨基联苯胺(DAB)增强剂和DAB分别呈色；以及同时用碱性磷酸酶(AP)和HRP双酶分别标记TH和Fos,再分别用5-溴-4-氯-3-吲哚磷酸盐(BCIP)和氯化硝基四氮唑蓝(NBT)对TH和DAB对Fos呈色,比较免疫双标时两种不同酶标记显色方法...

Effect of HRP and AP Labeling on Fos Expression in Catecholamine Neurons of the NTS of Rats During Restraint Water-immersion Stress

A dual immunohistochemical technique is used to detect the effect of horse radish peroxidase(HRP) and alkaline phosphatase(AP) labeling on Fos expression in catecholaminergic neurons of the neucleus of solitary tract(NTS) of rats during restraint water-immersion stress(RWIS).One method is that horse radish peroxidase(HRP) labeled Fos and TH in turn,and then using diaminobenzidine hydrochloride(DAB),intensified with 0.05% cobalt chloride and 0.05% nickel ammonium sulfate,to visualize Fos-immunoreactive neurons(Fos-IR),and DAB to TH-immunoreactive neurons(TH-IR).The other method is that HRP labeled Fos and alkaline phosphatase(AP) labeled TH neurons,and then using DAB and 5-bromo-4-chloro-3-indolylphosphate(BCIP)/nitroblue tetrazolium(NBT) to visualize Fos-IR and TH-IR respectively.Fos-IR nuclei,TH-IR,Fos and TH double labeled neurons evidently increase in stressed rats.The two methods all fit to reveal the phenotypic nature of the activated neurons.But using HRP labeling Fos and TH respectively would cost much more time and lead to a nonspecific immunoreactivity,and if the antibodies come from different species in origin,HRP/AP is a priority.