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Localization of high affinity choline transporter in the primate nervous system.

高亲和力胆碱转运蛋白在灵长类神经系统中的定位。

基本信息

批准号：
15500246
负责人：
MATSUMURA George
金额：
$ 1.98万
依托单位：
Kyorin University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2005
项目状态：
已结题

项目摘要

1.Localization in the central nervous systemUsing in situ hybridization and immunohistochemistry, we have determined the localization of high affinity choline transporter (CHT) messenger RNA and protein in the macaque monkey central nervous system. Signals for CHT mRNA and protein are localized in neurons in the motor nuclei of the cranial nerves (somatic motor and parasympathetic preganglionic), laterodorsal tegmental nucleus, parabrachial nucleus, pedunculopontine tegmental nucleus, globus pallidus, neostriatum, nucleus accumbens, nucleus of the diagonal band of Broca, nucleus basalis of Meynert, medial septal nucleus, and olfactory tubercle. Parasympathetic preganglionic neurons exhibited significantly lower density of CHT mRNA and protein than other positive neurons. In the spinal cord, motor neurons in the ventral horn showed both signals for CHT mRNA and protein. Putative gamma-motor neurons always showed strong immunoreactivity for CHT, while putative alpha-motor neurons showed … More wide variety of immunoreactivity intensities. Of putative alpha-motor neurons, neurons innervating principal dorsal muscles demonstrated higher content of CHT than neurons innervating limb muscles.2.Ultrastructural localization at the synapseTo determine whether CHT is located at the presynaptic membrane or at the membrane of synaptic vesicles, we analyzed the localization of CHT at the monkey neuromuscular junction using post-embedding immunocytochemistry with electronmicroscopy. Although immunoreactivity was weaker than reported in the rat, most of the immunoreactivity for CHT was located inside the nerve terminal rather than on the presynaptic membrane. The findings are compatible with the CHT localization on the vesicular membrane reported in the rat, but for the exact localization at the vesicular membrane, further study will be necessary to determine the optimal condition that preserve both the immunoreactivity for CHT and the structure of vesicular membrane. These electron microscopic studies were performed in collaboration with Professor Seiji Matsuda at Ehime University. Less

1.中枢神经系统定位利用原位杂交和免疫组化技术，我们确定了高亲和力胆碱转运蛋白（CHT）信使RNA和蛋白质在猕猴中枢神经系统中的定位。 CHT mRNA 和蛋白质的信号位于脑神经运动核（躯体运动神经节前和副交感神经节前）、背侧被盖核、臂旁核、桥脚被盖核、苍白球、新纹状体、伏隔核、布罗卡对角带核的神经元中， Meynert 基底核、内侧间隔核和嗅结节。副交感节前神经元的 CHT mRNA 和蛋白密度明显低于其他阳性神经元。在脊髓中，腹角的运动神经元同时显示 CHT mRNA 和蛋白质信号。假定的 γ 运动神经元总是对 CHT 显示出强烈的免疫反应性，而假定的 α 运动神经元则显示出各种各样的免疫反应强度。在假定的α运动神经元中，支配主要背肌的神经元比支配肢体肌肉的神经元表现出更高的CHT含量。2.突触的超微结构定位为了确定CHT是位于突触前膜还是突触小泡膜，我们分析了CHT在猴神经肌肉接头处的定位电子显微镜下的包埋后免疫细胞化学。尽管大鼠体内的免疫反应性比报道的要弱，但 CHT 的大部分免疫反应性位于神经末梢内部，而不是突触前膜上。这些发现与大鼠中报告的囊泡膜上的 CHT 定位相一致，但对于囊泡膜上的精确定位，需要进一步研究以确定保留 CHT 免疫反应性和囊泡膜结构的最佳条件。这些电子显微镜研究是与爱媛大学松田诚二教授合作进行的。较少的