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使用亚基特异性抗体测定大鼠脑和交感神经节中的神经元烟碱型乙酰胆碱受体α3亚基蛋白:区域和个体发育表达

Neuronal nicotinic acetylcholine receptor alpha3 subunit protein in rat brain and sympathetic ganglion measured using a subunit-specific antibody: regional and ontogenic expression.

作者信息

Yeh J J, Yasuda R P, Dávila-García M I, Xiao Y, Ebert S, Gupta T, Kellar K J, Wolfe B B

机构信息

Department of Pharmacology, Georgetown University School of Medicine, Washington DC, USA. Department of Pharmacology, Howard University, Washington DC, USA.

出版信息

J Neurochem. 2001 Apr;77(1):336-46. doi: 10.1046/j.1471-4159.2001.t01-1-00259.x.

Abstract

A synthetic peptide corresponding to the C-terminus of the alpha 3 subunit of the rat neuronal nicotinic acetylcholine receptor (nAChR) was used to generate a rabbit polyclonal alpha 3 antibody. The specificity of this antibody was characterized by immunoblotting, immunohistochemical and immunoprecipitation techniques. Using this antibody, the relative densities of the alpha 3 subunit were quantitatively determined in different brain regions and in superior cervical ganglion (SCG). Among these regions, SCG, interpeduncular nucleus (IPN) and pineal gland showed the highest levels of alpha 3 protein expression. Habenula and superior colliculi had intermediate levels of expression. Low levels were found in cerebral cortex, hippocampus and cerebellum. The ontogenic profile of the alpha 3 subunit in the SCG was also determined. The alpha 3 protein level is low at postnatal day (P 1), but increases rapidly during the first seven postnatal days. This level then plateaus and remains stable through postnatal day 35. These findings suggest that neuronal nAChRs containing the alpha 3 subunit participate in important roles in specific regions of the rat brain and the SCG.

摘要

一种与大鼠神经元烟碱型乙酰胆碱受体(nAChR)α3亚基C末端对应的合成肽被用于制备兔抗α3多克隆抗体。该抗体的特异性通过免疫印迹、免疫组织化学和免疫沉淀技术进行表征。使用此抗体,在不同脑区和颈上神经节(SCG)中定量测定了α3亚基的相对密度。在这些区域中,SCG、脚间核(IPN)和松果体显示出最高水平的α3蛋白表达。缰核和上丘表达水平中等。在大脑皮层、海马和小脑中发现水平较低。还确定了SCG中α3亚基的个体发育情况。α3蛋白水平在出生后第1天(P1)较低,但在出生后的前7天迅速增加。然后该水平趋于平稳,并在出生后第35天保持稳定。这些发现表明,含有α3亚基的神经元nAChR在大鼠脑和SCG的特定区域中发挥重要作用。

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