Giaid A, Gibson S J, Herrero M T, Gentleman S, Legon S, Yanagisawa M, Masaki T, Ibrahim N B, Roberts G W, Rossi M L
Department of Histochemistry, Royal Postgraduate Medical School, Hammersmith Hospital, London, UK.
Histochemistry. 1991;95(3):303-14. doi: 10.1007/BF00266781.
The distribution of endothelin mRNA and immunoreactivity in the human brain was investigated using the technique of in situ hybridization and immunocytochemistry. Cryostat sections from 22 cases of neurologically normal adult human brain, collected 3-7 h post-mortem were hybridized with 35S-labelled complementary (c)RNA probes prepared from the 3' non-coding region of endothelin-1 cDNA, and the chromosomal genes encoding endothelin-2 and -3. In situ hybridization with all three cRNA probes revealed labelled neuronal cell bodies in laminae III-VI of the parietal, temporal and frontal cortices. Labelled cells were also seen, scattered throughout the para- and periventricular, supraoptic and lateral hypothalamic nuclei, the caudate nucleus, amygdala, hippocampus, basal nucleus of Meynert, substantia nigra, raphe nuclei, Purkinje cell layer of the cerebellum and in the dorsal motor nuclei of the vagus of the medulla oblongata. The distribution of neurones immunoreactive to endothelin was similar to that of endothelin mRNA, although fewer immunoreactive cells throughout the brain, were noted. Immunoreactive fibres were present mainly in the cortex and hypothalamus, and to a lesser extent in the brain stem. Combined in situ hybridization and immunocytochemistry on the same section revealed the presence of endothelin-1 mRNA and immunoreactivity in the same cortical neuronal cell. Colocalisation studies in the cortex revealed endothelin-1 mRNA and immunoreactivity in a number of cells which also expressed neuropeptide Y mRNA and immunoreactivity. In the hypothalamus and basal nucleus of Meynert endothelin immunoreactivity was colocalised to a subset of neurophysin- and galanin-immunoreactive cell bodies respectively. Endothelin mRNA and immunoreactivity was also seen in some blood vessel endothelial cells. The findings of endothelin mRNAs and immunoreactivity in heterogenous neuronal populations further emphasises the potential role of endothelin as a neuropeptide, probably having diverse actions in the nervous system of man.
采用原位杂交和免疫细胞化学技术研究了内皮素mRNA和免疫反应性在人脑内的分布情况。对22例神经功能正常的成年人脑进行尸检，在死后3 - 7小时采集冰冻切片，用从内皮素-1 cDNA的3'非编码区制备的35S标记互补（c）RNA探针以及编码内皮素-2和-3的染色体基因进行杂交。用所有三种cRNA探针进行原位杂交，均在顶叶、颞叶和额叶皮质的III - VI层发现了标记的神经元细胞体。在室旁核和室周核、视上核、下丘脑外侧核、尾状核、杏仁核、海马、迈内特基底核、黑质、中缝核、小脑浦肯野细胞层以及延髓迷走神经背运动核中也可见到散在的标记细胞。对内皮素免疫反应阳性的神经元分布与内皮素mRNA的分布相似，不过在全脑范围内免疫反应阳性细胞较少。免疫反应性纤维主要存在于皮质和下丘脑，在脑干中含量较少。在同一切片上进行原位杂交和免疫细胞化学联合检测，发现同一皮质神经元细胞中存在内皮素-1 mRNA和免疫反应性。在皮质进行的共定位研究显示，内皮素-1 mRNA和免疫反应性存在于一些同时表达神经肽Y mRNA和免疫反应性的细胞中。在下丘脑和迈内特基底核中，内皮素免疫反应性分别与一部分神经垂体素和甘丙肽免疫反应阳性的细胞体共定位。在一些血管内皮细胞中也可见到内皮素mRNA和免疫反应性。内皮素mRNA和免疫反应性在多种神经元群体中的发现，进一步强调了内皮素作为一种神经肽的潜在作用，它可能在人类神经系统中发挥多种作用。
