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成年大鼠脑中已鉴定出的胆碱能神经元富含生长相关蛋白43(GAP - 43)信使核糖核酸(mRNA):一项双重原位杂交研究

Identified cholinergic neurones in the adult rat brain are enriched in GAP-43 mRNA: a double in situ hybridisation study.

作者信息

Augood S J, Arbuthnott G W, Emson P C

机构信息

Department of Neurobiology, Babraham Institute, Cambridge, UK.

出版信息

J Chem Neuroanat. 1995 Jul;9(1):17-26. doi: 10.1016/0891-0618(95)00059-g.

Abstract

The cellular expression of growth associated protein-43 mRNA by identified choline acetyl transferase mRNA positive cells was investigated in the mature rat brain using a combined radioactive and non-radioactive in situ hybridisation technique. Cellular sites of growth associated protein-43 mRNA were detected using a 35S-oligonucleotide while choline acetyl transferase mRNA positive neurones were identified using two alkaline phosphatase-labelled probes. In the cholinergic cells of the corpus striatum, basal forebrain and laterodorsal tegmental nucleus a specific growth associated protein-43 hybridisation signal (silver grains) was detected, demonstrating that these choline acetyl transferase mRNA positive cells are enriched in growth associated protein-43 gene transcripts. By contrast, the large cholinergic cells of the motor nucleus of the trigeminal nerve did not express growth associated protein-43 mRNA. Quantification of the growth associated protein-43 hybridisation signal expressed by identified choline acetyl transferase mRNA positive cells showed regional variations in the relative cellular abundance of this transcript; cholinergic cells in the laterodorsal tegmental nucleus and corpus striatum expressed the strongest cellular hybridisation signal. Mean cross-sectional somatic area measurements of these growth associated protein-43/cholinergic positive cells confirmed the identity of these neurones as belonging to the cholinergic phenotype. A strong 35S-growth associated protein-43 hybridisation signal was detected also in numerous other non-choline acetyl transferase mRNA positive nerve cells in other regions of the brain, although the chemical phenotypes of these neurones were not determined. Our data reveal that expression of the growth-associated protein GAP-43 is maintained in identified cholinergic neurones in the postnatal rat brain, suggesting that this protein may subserve important functions in cholinergic and other neurones of the adult mammalian brain.

摘要

采用放射性与非放射性原位杂交技术相结合的方法,在成年大鼠脑中研究了已鉴定的胆碱乙酰转移酶mRNA阳性细胞中生长相关蛋白-43 mRNA的细胞表达情况。利用35S-寡核苷酸检测生长相关蛋白-43 mRNA的细胞定位,同时使用两种碱性磷酸酶标记的探针鉴定胆碱乙酰转移酶mRNA阳性神经元。在纹状体、基底前脑和外侧背侧被盖核的胆碱能细胞中检测到特异性的生长相关蛋白-43杂交信号(银颗粒),表明这些胆碱乙酰转移酶mRNA阳性细胞富含生长相关蛋白-43基因转录本。相比之下,三叉神经运动核的大型胆碱能细胞不表达生长相关蛋白-43 mRNA。对已鉴定的胆碱乙酰转移酶mRNA阳性细胞所表达的生长相关蛋白-43杂交信号进行定量分析,结果显示该转录本的相对细胞丰度存在区域差异;外侧背侧被盖核和纹状体中的胆碱能细胞表达最强的细胞杂交信号。对这些生长相关蛋白-43/胆碱能阳性细胞的平均横截面积测量结果证实了这些神经元属于胆碱能表型。在脑的其他区域的许多其他非胆碱乙酰转移酶mRNA阳性神经细胞中也检测到强烈的35S-生长相关蛋白-43杂交信号,尽管这些神经元的化学表型尚未确定。我们的数据表明,出生后大鼠脑中已鉴定的胆碱能神经元中生长相关蛋白GAP-43的表达得以维持,这表明该蛋白可能在成年哺乳动物脑的胆碱能及其他神经元中发挥重要作用。

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