EXPRESSION OF NGF RECEPTOR IN THE RAT FOREBRAIN DETECTED WITH INSITU HYBRIDIZATION AND IMMUNOHISTOCHEMISTRY
EXPRESSION OF NGF RECEPTOR IN THE RAT FOREBRAIN DETECTED WITH INSITU HYBRIDIZATION AND IMMUNOHISTOCHEMISTRY
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DOI：
10.1016/0169-328x(89)90073-9
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发表时间：
1989-12-01
期刊：
影响因子：
--
通讯作者：
PFAFF, DW
中科院分区：
文献类型：
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作者：
GIBBS, RB;MCCABE, JT;PFAFF, DW
应用原位杂交和免疫组织化学技术检测神经生长因子受体(NGF)受体mRNA和蛋白在成年大鼠基底前脑中的表达。NGF受体在内侧隔核、Broca斜角带和Meynert基底核的细胞内均有表达。对照表明，杂交信号不是由于探针与异源RNA或其他分子的非特异性结合所致。正如预期的那样，含有NGF受体mRNA的细胞的分布与NGF受体免疫反应细胞在这些区域的分布有很好的相关性。这些数据扩展了以前的工作，即这些区域的神经元表达NGF受体mRNA，并产生功能性的NGF受体。NGF受体在下丘脑弓状核、脑底和顶盖上的软脑膜以及脑室侧壁的室管膜内也有表达。在下丘脑外侧和腹外侧纹状体也观察到少量弱染色神经元。相反，通过原位杂交，在任何脑膜、室管膜或下丘脑组织中都没有检测到NGF受体mRNA。交叉连接/免疫沉淀分析显示，NGF受体在背侧上丘、下丘脑腹内侧部和上面的脑膜组织中存在正常的膜结合受体，证明在这些区域观察到的染色不是与免疫组织化学相关的特定人工制品。这些区域的杂交缺失可能反映了NGF受体mRNA的水平太低，无法用正在使用的原位杂交方法检测到。或者，这种染色可能代表了这些区域的神经传入，其细胞体位于其他地方，其终末含有NGF受体蛋白。
The expression of nerve growth factor (NGF) receptor mRNA and NGF receptor protein was examined in the adult rat basal forebrain using in situ hybridization and immunohistochemical techniques. NGF receptor mRNA and protein were detected within cells in teh medial septum, diagonal band of Broca, and nucleus basalis of Meynert. Controls showed that the hybridization signal was not due to nonspecific binding of the probe to heterologous RNAs or other molecules. As expected, the distribution of NGF receptor mRNA-containing cells correlated nicely with the distribution of NGF receptor immunoreactive cells in each of these areas. These data extend previous work which suggests that neurons in these areas express the NGF receptor mRNA and manufacture functional NGF receptors. NGF receptor immunoreactivity was also detected in the arcuate nucleus of the hypothalamus, in the leptomeninges at the base of the brain and overlying the tectum, and within ependymal regions along the lateral walls of the cerebral ventricles. A few weakly stained neurons in the lateral hypothalamus and ventrolateral striatum werealso consistently observed. In contrast, NGF receptor mRNA was not detected within any meningial, ependymal, or hypothalamic tissues using in situ hybridization. A cross-linking/immunoprecipitation assay demonstrated normal, membrane-bound NGF receptors within extracts of dorsal superior colliculus, ventromedial hypothalamic, and overlying meningial tissues, proving that the staining observed in these areas was not a on-specific artifact associated with the immunohistochemistry. The lack of hybridization in these areas may reflect levels of NGF receptor mRNA which are too low to be detected by the in situ hybridization methods being used. Alternatively, the staining may represent innervation of these areas by afferents whose cell bodies are located elsewhere, and whose terminals contain the NGF receptor protein.