DISTRIBUTION OF SOMATOSTATIN MESSENGER-RNA IN THE RAT NERVOUS-SYSTEM AS VISUALIZED BY A NOVEL NONRADIOACTIVE INSITU HYBRIDIZATION HISTOCHEMISTRY PROCEDURE
DISTRIBUTION OF SOMATOSTATIN MESSENGER-RNA IN THE RAT NERVOUS-SYSTEM AS VISUALIZED BY A NOVEL NONRADIOACTIVE INSITU HYBRIDIZATION HISTOCHEMISTRY PROCEDURE
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DOI：
10.1016/0306-4522(90)90388-k
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发表时间：
1990-01-01
期刊：
影响因子：
3.3
通讯作者：
EMSON, PC
中科院分区：
文献类型：
--
作者：
KIYAMA, H;EMSON, PC
用一种新开发的高度敏感的非放射性原位杂交组织化学方法，详细研究了大鼠脑和感觉神经节中前生长抑素原mRNA的细胞定位。用碱性磷酸酶标记的反义寡核苷酸探针检测生长抑素mRNA。这种探针很容易证明生长抑素基因的表达在整个大鼠中枢神经系统具有非常高的对比度和良好的细胞定位。结果，我们在许多以前未显示含有mRNA信号的CNS区域中观察到大量生长抑素mRNA阳性细胞。该方法在副嗅球的僧帽细胞层、主嗅球的肾小球层、外侧隔的背侧部、上级丘、下丘的浅灰质层、耳蜗前腹侧核、小脑的颗粒层和浦肯野细胞层、延髓和脊髓的胶状质中检测到大量的生长抑素mRNA阳性细胞。所有使用放射性标记的原位探针进行信号检测以前相当困难的领域。本方法的主要优点包括信号的非常精确的细胞分辨率、快速的反应时间和低背景。本方法的灵敏度似乎至少相当于大多数免疫细胞化学程序，比大多数同位素原位杂交方法更敏感。
The cellular localization of preprosomatostatin mRNA in the rat brain and sensory ganglia has been examined in detail using a newly developed highly sensitive non-radioactive in situ hybridization histochemistry procedure. An alkaline phosphatase labelled anti-sense 30mer oligodeoxynucleotide probe was used for detection of somatostatin mRNA. This probe readily demonstrated somatostatin gene expression throughout the rat CNS with very high contrast and good cellular localization. As a result, we visualized numerous somatostatin mRNA-positive cells in many CNS areas which had previously not been shown to contain a mRNA signal. This method detected a number of somatostatin mRNA-positive cells, in the mitral cell layer of accessory olfactory bulb, the glomerular layer of the main olfactory bulb, the dorsal part of the lateral septum, superficial gray layer of superior colliculus, inferior colliculus, anterior ventral cochlear nucleus, granular layer and Purkinje cell layer of cerebellum, and substantia gelatinosa of medulla and spinal cord, all areas where signal detection using radiolabelled in situ probes has previously been rather difficult. The principle advantages of the present method include the very precise cellular resolution of signal, the rapid reaction time and low background. The sensitivity of the present method seems to be at least equivalent to most immunocytochemical procedures and more senstive than most isotopic in situ hybridization methods.