原位杂交组织化学与免疫组织化学结合法的研究

A STUDY ON THE COMBINATION OF IN SITU HYBRIDIZATION HISTOCHEMISTRY WITH IMMUNOHISTOCHEMISTRY

介绍一种用原位杂交组织化学与免疫组织化学结合法在同一切片中显示生长抑素mRNA和相对应的生长抑素.同时还比较了六种不同固定液对原位杂交组织化学和原位杂交组织化学与免疫组织化学结合法的影响.结果表明Zamboni氏固定液用于原位杂交组织化学与免疫组织化学结合法最佳,而4%多聚甲醛磷酸缓冲液用单纯原位杂交最佳.原位杂交组织化学与免疫组织化学结合法显示的下丘脑室周核生长抑素免疫反应阳性神经元均含有相应生长抑素mRNA,而且多数神经元生长抑素基因的转录量与生长抑素的翻译量成正比.

The combination of in situ hybridization histochemistry with immunocytochemistry was studied to demonstrate somatostatin immunoreactivity and somatostatin mRNA on the same section of the rat hypothalamus.A comparison was made to assess the effects of 6 fixatives on in situ hybridization histochemistry and its comination with immunohistochemistry.The results showed that Zamboni's fixative was the best for the combination of in situ hybridization histochemistry with immunohistochemistry while 4% paraformaldehyde-phosphate buffer was best for in situ hybridization histochemistry.All,somatostatin immunoreactive neurons demonstrated by combining in situ hybridization histochemistry with immunohistochemistry contained silver granules of somatostatin mRNA in the periventricular nucleus of the rat hypothalamus.The amount of somatostatin gene transcription in proportional that of somatostatin translation from its mRNA.