Immunohistochemical localisation of the 5-HT2C receptor protein in the rat CNS
Immunohistochemical localisation of the 5-HT2C receptor protein in the rat CNS
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DOI：
10.1016/s0028-3908(99)00086-6
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发表时间：
2000-01-01
影响因子：
4.7
通讯作者：
Fone, KCF
中科院分区：
文献类型：
--
作者：
Clemett, DA;Punhani, T;Fone, KCF
5-HT 2C受体mRNA在人和大鼠CNS中具有广泛的分布，但是缺乏特异性高亲和力配体使得受体的放射自显影定位困难。通过制备针对大鼠5-HT 2C受体蛋白的多克隆抗体，本研究报告了该受体在大鼠中枢神经系统中的免疫组织化学分布。Sephadex纯化的5-HT 2C抗血清在从几个大鼠脑区域制备的膜的Western印迹中显现出单一的免疫阳性条带（54 kDa），并在用h5-HT 2C cDNA转染的HEK 293细胞中引起强烈的膜免疫荧光，这两者都通过与抗原肽序列（200-300 μ M）孵育而减弱。5-HT 2C样免疫反应位于整个中枢神经系统的神经元。最丰富的5-HT 2C样免疫反应细胞体在前嗅核，内侧和闰杏仁核，海马CA 1至CA 3层，laterodoral和外侧膝状体丘脑核，尾壳核和几个地区的皮质（包括梨状和额叶），与此受体位于突触后的多巴胺能神经元。在中缝背核中也发现了免疫阳性神经元，提示5-HT 2C受体可能存在于某些肾上腺素能神经元上。5-HT 2C样免疫反应性的总体分布补充了以前的研究结果与传统的放射性配体，并同意与5-HT 2C受体mRNA的报告水平。(C)1999 Elsevier Science Ltd.保留所有权利。
5-HT2C receptor mRNA has a widespread distribution in the human and rat CNS but the absence of a specific high affinity ligand has made autoradiographic localisation of the receptor difficult. By raising polyclonal antibodies against the rat 5-HT2C receptor protein this study reports the immunohistochemical distribution of this receptor in the rat CNS. A sephadex purified 5-HT2C antiserum visualised a single immunopositive band (54 kDa) in Western blots of membranes prepared from several rat brain regions and caused intense membrane immunofluorescence in HEK 293 cells transfected with h5-HT2C cDNA, which were both attenuated by incubation with the antigenic peptide sequence (200-300 mu M). 5-HT2C-like immunoreactivity was located on neurones throughout the CNS. The most abundant 5-HT2C-like immunoreactive cell bodies were in the anterior olfactory nucleus, medial and intercalated amygdaloid nuclei, hippocampus layers CA1 to CA3, laterodorsal and lateral geniculate thalamic nuclei, caudate-putamen and several areas of the cortex (including piriform and frontal), consistent with this receptor being located postsynaptic to serotonergic neurones. Immunopositive neurones were also found in the dorsal raphe, suggesting that 5-HT2C receptors may be on some serotonergic neurones. The overall distribution of 5-HT2C-like immunoreactivity complements previous findings with conventional radioligands and agrees well with reported levels of 5-HT2C receptor mRNA. (C) 1999 Elsevier Science Ltd. All rights reserved.