Neurochemical properties of enkephalinergic neurons in lumbar spinal dorsal horn revealed by preproenkephalin-green fluorescent protein transgenic mice
Neurochemical properties of enkephalinergic neurons in lumbar spinal dorsal horn revealed by preproenkephalin-green fluorescent protein transgenic mice
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前脑啡肽原-绿色荧光蛋白转基因小鼠揭示腰椎背角脑啡肽能神经元的神经化学特性
DOI：
10.1111/j.1471-4159.2010.06715.x
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发表时间：
2010-06-01
影响因子：
4.7
通讯作者：
Wu, Sheng-Xi
中科院分区：
文献类型：
--
作者：
Huang, Jing;Chen, Jing;Wu, Sheng-Xi
脑啡肽（ENK）与脊髓的伤害性传递有关，但由于难以理想地观察ENK能神经元，其功能作用尚不清楚。因此，我们开发了脑啡肽前绿色荧光蛋白转基因小鼠，以更好地识别ENKergic神经元。实时逆转录聚合酶链反应（RT-PCR）、免疫组织化学和原位杂交技术首次证实了转基因操作的成功及其在脊髓ENKergic神经元显示中的应用。脊髓I、II、III和IV-VI层的ENKergic神经元在背角神经元中的比例分别为15.8 +/- 3.1%、39.5 +/- 3.3%、11.8 +/- 1.9%和10.7 +/- 2.1%。然后与其他分子进行双重标记，以进一步阐明脊髓ENKergic神经元的神经化学特性。42.9 +/- 2.8%的ENKergic神经元中存在GABA，主要分布在I-III层。ENKergic神经元中细小白蛋白、钙黄蛋白、钙结合蛋白和神经元一氧化氮合酶阳性细胞的比例分别为5.2 +/- 0.7%、42.6 +/- 2.3%、25.8 +/- 2.2%和11.1 +/- 1.6%。与先前用ENK抗体标记获得的结果相比，这一新生小鼠系可以成为研究特定脊髓ENKergic神经元群体的可靠工具。
P>Enkephalin (ENK) has been implicated in nociceptive transmission in the spinal cord while its functional role is not clear because of difficulties in ideally visualizing ENKergic neurons. We thus developed preproenkephalin-green fluorescent protein transgenic mice to better identify ENKergic neurons. Real-time reverse transcriptase-polymerase chain reaction (RT-PCR) together with immunohistochemistry and in situ hybridization were first employed to confirm the successful transgenic manipulation and its application in showing spinal ENKergic neurons. The proportions of ENKergic neurons in the spinal cord laminae I, II, III and IV-VI among dorsal horn neurons were 15.8 +/- 3.1%, 39.5 +/- 3.3%, 11.8 +/- 1.9% and 10.7 +/- 2.1%, respectively. Double labeling with other molecules was then performed to further clarify the neurochemical properties of spinal ENKergic neurons. GABA was found to exist in 42.9 +/- 2.8% of ENKergic neurons that were mainly located in lamina I-III. The proportions of parvalbumin-, calretinin-, calbindin- and neuronal nitric oxide synthase-positive cells among the ENKergic neurons were 5.2 +/- 0.7%, 42.6 +/- 2.3%, 25.8 +/- 2.2% and 11.1 +/- 1.6%, respectively. Compared with previously findings obtained with ENK antibody labeling, this line of newly generated mice can be a reliable tool for the study of specific spinal ENKergic neuronal population.