Kiyama H, Morita Y, Noguchi K, Wang Y, Nakanishi S, Shiotani Y, Tohyama M
Department of Neuroanatomy, Osaka University Medical School, Japan.
J Chem Neuroanat. 1988 May-Jun;1(3):125-32.
The localization of gamma-preprotachykinin A mRNAs in the rat trigeminal ganglion was demonstrated by in situ hybridization histochemistry using the 32P and 35S labelled gamma-preprotachykinin A complementary DNA. In situ hybridization using 32P allowed shorter exposure times, whereas higher resolution of the hybridization signal on both film and emulsion autoradiograms was obtained using 35S. Preprotachykinin A mRNA detected by the gamma-preprotachykinin A probe was localized in about 15 per cent of the trigeminal ganglion cells, most of which were small or medium sized. Immunohistochemical studies using anti-substance P antibodies demonstrated that 15-20 per cent of total trigeminal ganglion cells were positive. These cells were small or medium sized. The result of immunohistochemistry coincided well with that of in situ hybridization histochemistry. The present study showed that the cellular localization of preprotachykinin A mRNA could be analysed by in situ hybridization histochemistry.
利用32P和35S标记的γ-前速激肽原A互补DNA，通过原位杂交组织化学法证实了γ-前速激肽原A mRNA在大鼠三叉神经节中的定位。使用32P进行原位杂交可缩短曝光时间，而使用35S可在胶片和乳胶放射自显影片上获得更高分辨率的杂交信号。用γ-前速激肽原A探针检测到的前速激肽原A mRNA定位于约15%的三叉神经节细胞中，其中大多数为小或中等大小。使用抗P物质抗体的免疫组织化学研究表明，三叉神经节细胞总数的15%-20%呈阳性。这些细胞为小或中等大小。免疫组织化学结果与原位杂交组织化学结果吻合良好。本研究表明，可通过原位杂交组织化学法分析前速激肽原A mRNA的细胞定位。
