PACKAGING AND AXONAL TRANSPORT OF THREE MACROMOLECULES
三种大分子的包装和轴突运输
基本信息
- 批准号:2266074
- 负责人:
- 金额:$ 19.57万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:1989
- 资助国家:美国
- 起止时间:1989-08-01 至 1996-07-31
- 项目状态:已结题
- 来源:
- 关键词:Torpedo acetylcholinesterase alternatives to animals in research axon cell membrane electron microscopy endoplasmic reticulum enzyme linked immunosorbent assay extracellular matrix laboratory rat macromolecule membrane structure neuroanatomy neuronal transport neurons potassium channel protein biosynthesis protein structure function protein transport radioimmunoassay sodium channel sodium potassium exchanging ATPase soma voltage gated channel
项目摘要
This proposal details a research program designed to localize three
specific macromolecules within intracellular membrane systems of both the
cell body and the axon. Localization of these macromolecules will
elucidate the involvement of specific somal membrane systems in the
packaging of proteins destined for different target areas of the plasma
membrane or extracellular space and the involvement of axonal membrane
systems in axonal transport. The three target macromolecules to be
studied are; 1) the Na+ +K+ ATPase; 2) the voltage dependent SODIUM
CHANNEL; and 3) ACETYLCHOLINESTERASE (AChE). These macromolecules (their
individuals constituent subunits or distinct molecular forms) will be
localized with the aid of specific antibody probes visualized by light
and electron microscopy using gold, ferritin, peroxidase, or fluorescent
antibody conjugates. The membrane systems under study are both those of
the cell body and of the axon. In the cell body, those involved in the
biosynthesis and packaging of membrane proteins or secretion products;
e.g., the rough endoplasmic reticulum (RER), the smooth endoplasmic
reticulum (SER), and the Golgi apparatus. In the axon, there are at least
three morphologically and functionally distinct membrane systems. One of
these is clearly involved in anterograde rapid axonal transport, another
in retrograde transport and the third which does not appear to be rapidly
transported and whose function or relationship to the other two is
unknown. These planned investigations into the biosynthesis and transport
of all three of these macromolecules are now enabled by the recent
production and characterization of specific antibodies to 10 the Na+ +K+
ATPase and its alpha and beta subunits from rat and eel (Electrophorus
electricus) tissues, 2) the Na+ channel from eel, and 3) two of the major
molecular forms of AChE from Torpedo. We have already proven antibodies
to each of these macromolecules suitable for use in high resolution
immunoelectron microscopic studies, immunofluorescent studies, or as
biochemical probes with techniques including Western blotting, enzyme
linked immunoadsorbent assays or radioimmunoassays. Species and tissue
specificity of antibodies have been examined. All are capable of
recognizing neuronal forms of each antigen and in most cases cytoplasmic
labeling capacity has been already confirmed.
该提案详细介绍了一项研究计划,旨在将三个
细胞内膜系统中的特定大分子
细胞体和轴突。这些大分子的本地化将
阐明特定的胞体膜系统参与
去往血浆不同靶区的蛋白质包装
膜或细胞外间隙与轴突膜的受累
轴突运输系统。这三个目标大分子是
研究有:1)Na++K+-ATPase;2)电压依赖性钠
3)乙酰胆碱酯酶(AChE)。这些大分子(他们的
个人组成亚单位或不同的分子形式)将是
借助光显示的特异性抗体探针进行定位
使用金、铁蛋白、过氧化物酶或荧光的电子显微镜
抗体偶联。正在研究的膜系统都是
细胞体和轴突。在细胞体中,那些参与
膜蛋白或分泌物的生物合成和包装;
例如,粗面内质网(RER)、光滑内质网
网(SER)和高尔基体。在轴突中,至少有
三个在形态和功能上截然不同的膜系统。其中之一
这些明显参与了顺行快速轴突运输,另一种
在逆行运输中,第三个看起来不是很快
其功能或与其他两个人的关系是
未知。这些计划对生物合成和运输的调查
所有这三个大分子现在都能通过最近的
抗10-Na++K+特异性抗体的制备及鉴定
大鼠和鳗鱼的ATPase及其α、β亚基
电)组织,2)鳗鱼的Na+通道,以及3)两个主要的
来自鱼雷的乙酰胆碱酯酶的分子形式。我们已经证实了抗体
适用于高分辨率的每一种大分子
免疫电子显微镜研究、免疫荧光研究或AS
带有蛋白质印迹、酶等技术的生化探针
联用免疫吸附测定法或放射免疫测定法。物种和组织
对抗体的特异性进行了检测。所有人都有能力
识别每个抗原的神经元形式,在大多数情况下是胞浆
贴标能力已经得到确认。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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Mark H Ellisman其他文献
Mark H Ellisman的其他文献
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{{ truncateString('Mark H Ellisman', 18)}}的其他基金
200keV, Energy Filtered, Intermediate-High Voltage Transmission Electron Microscope(IVEM)"
200keV、能量过滤、中高压透射电子显微镜(IVEM)"
- 批准号:
10642585 - 财政年份:2023
- 资助金额:
$ 19.57万 - 项目类别:
Scalable electron tomography for connectomics
用于连接组学的可扩展电子断层扫描
- 批准号:
10410742 - 财政年份:2022
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逆转阿尔茨海默病中的小胶质细胞炎症和线粒体功能障碍
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10607455 - 财政年份:2022
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National Center for Microscopy and Imaging Research: A BRAIN Technology Integration and Dissemination Resource
国家显微镜和成像研究中心:大脑技术集成和传播资源
- 批准号:
10334513 - 财政年份:2021
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National Center for Microscopy and Imaging Research: A BRAIN Technology Integration and Dissemination Resource
国家显微镜和成像研究中心:大脑技术集成和传播资源
- 批准号:
10544010 - 财政年份:2021
- 资助金额:
$ 19.57万 - 项目类别:
National Center for Microscopy and Imaging Research: A BRAIN Technology Integration and Dissemination Resource
国家显微镜和成像研究中心:大脑技术集成和传播资源
- 批准号:
10116087 - 财政年份:2021
- 资助金额:
$ 19.57万 - 项目类别:
The National Center for Microscopy and Imaging Research, a Community-wide Scientific Resource
国家显微镜和成像研究中心,社区范围的科学资源
- 批准号:
10399337 - 财政年份:2020
- 资助金额:
$ 19.57万 - 项目类别:
Advancing Multi-Color EM via Direct Detector-enabled 4D-STEM
通过支持直接检测器的 4D-STEM 推进多色 EM
- 批准号:
10031737 - 财政年份:2020
- 资助金额:
$ 19.57万 - 项目类别:
Advancing Multi-Color EM via Direct Detector-enabled 4D-STEM
通过支持直接检测器的 4D-STEM 推进多色 EM
- 批准号:
10795540 - 财政年份:2020
- 资助金额:
$ 19.57万 - 项目类别:
The National Center for Microscopy and Imaging Research, a Community-wide Scientific Resource
国家显微镜和成像研究中心,社区范围的科学资源
- 批准号:
10212509 - 财政年份:2020
- 资助金额:
$ 19.57万 - 项目类别:
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