辣根过氧化物酶(Horseradish Peroxidase 简称 HRP)

最早由 Richard、Graham 和Karnovsky

(1968年)等人用来研究肾近端小管的早期吸收情况

将 HRP 注入鼠静脉内

间隔一定时间进行观察。在70年代初期

Kristensson 是将 HRP 用于神经系统联系的创始人

他的第一部分工作是在周缘神经腓肠肌内注射 HRP 而观察中枢(前角运动细胞)的 HRP 阳性标记细胞。其第二部分工作是将 HRP 注入舌肌观察舌下神经核内的标The horseradish peroxidase(HRP)method was established at the begining of 1970's.Using the HRP method for the observation of the segmentation of sympathetic afferent neuronsof the stomach

we found it suitable that the value(RZ)of HRP was over 2.5

concentration 10%and the total injected volume 80-100μl.The injection speed was 1 μl per minute.The duration of survival of the experimental animals for observation varied according to theirsize.For big animals(cat

rabbit)four to five days were needed.The fixation fluid used was only2% paraformaldehyde dissolved in phosphate acid buffer with pH 7.2-7.3.The perfusion should beperformed under 0°-4°C

so that the activity of HRP could be kept.In staining

a concentration of0

05 3

3

-diaminobenzidine(DAB)was used and twice staining would be preferred.First

the sectionswere incubated in DAB solution in aconstant temperature of 37℃ for 30 min.then H_2O_2 was addedinto the DAB solution under room temparature(18°-20°)for 30 min.The dark brown granulae inthe ner cells were clearly seen after the above procedure had been applied.The result showed that the above mentioned method was effective in the tracing back to the rela-tion of the peripheral nervous system.