NEUROBIOLOGIC STUDIES OF NEURONS AND GLIA IN CELL CULTURE
细胞培养中神经元和神经胶质细胞的神经生物学研究
基本信息
- 批准号:3778505
- 负责人:
- 金额:--
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:
- 资助国家:美国
- 起止时间:至
- 项目状态:未结题
- 来源:
- 关键词:acetylcholine action potentials aprotinin axon calcium flux chemical kinetics chick embryo electrophysiology glia growth cones hirudins horseradish peroxidase innervation muscle cells neurobiology neurons northern blottings optic chiasmas optic nerve optic tract protease inhibitor retinal ganglion synapses thrombin tissue /cell culture
项目摘要
Activity-dependent synapse elimination can be demonstrated in a
compartmental tissue culture system in which two populations of
cholinergic neurons converge on and innervate a common target
population of muscle cells. This synapse elimination can be blocked by
the inhibitor of serine and thiol proteases, leupeptin, at concentrations
of 10 nM and greater. The more specific blocker of serine proteases,
aprotinin, fails to block elimination as does the thiol protease blocker
cystatin. Hirudin is a highly specific inhibitor of the serine protease,
thrombin and hirudin does block synapse elimination at 1 nM
concentration (aprotinin does not block thrombin). Protease nexin I
(PNI), a naturally occurring protease inhibitor (identical with glial
derived nexin or GDN) also blocks thrombin and prevents activity-
dependent synapse elimination. These data strongly implicate thrombin
in the process of synapse elimination. Kinetic analysis of internal
calcium, [Ca++]i,, shows that single action potentials produce a larger
increase in [Ca++]i than do the individual action potentials of rapid (10
Hz) pulse trains. The peak [Ca++]i reached during repetitive bursts of
action potentials, however, is much higher than with single action
potentials. These kinetic data are useful in interpreting the mechanisms
coupling electrical activity to gene expression. Retinal ganglion cells
(RGC) in living chick embryos can be labelled with horseradish
peroxidase. The growth cones of the axons of these RGC are visualized
with this technique and exhibit dramatic changes at different places in
the optic pathway (nerve, chiasm and tract). This preparation should
allow evaluation of the effects of different manipulations of the visual
system (electrical stimulation, protease inhibitors, cell adhesion molecule
antibodies) upon growth cone morphology and axonal projection
patterns.
活动依赖性突触消除可以在
分室组织培养系统,其中两个群体的
胆碱能神经元聚集并支配共同的靶点
肌肉细胞群。 这种突触消除可以被以下物质阻断:
丝氨酸和巯基蛋白酶的抑制剂亮抑酶肽,
10 nM或更高。 丝氨酸蛋白酶的更特异性阻断剂,
抑肽酶不能像巯基蛋白酶阻断剂那样阻断消除
胱抑素 水蛭素是丝氨酸蛋白酶的高度特异性抑制剂,
凝血酶和水蛭素在1 nM时确实阻断突触消除
浓度(抑肽酶不阻断凝血酶)。 蛋白酶连接蛋白I
(PNI),一种天然存在的蛋白酶抑制剂(与神经胶质
衍生的连接蛋白或GDN)也阻断凝血酶并阻止活性-
依赖性突触消除 这些数据强烈暗示凝血酶
在突触消除的过程中。 内部动力学分析
钙,[Ca++]i,表明单个动作电位产生较大的
[Ca++]i的增加比快速(10
Hz)脉冲序列。 [Ca ~(++)]i在重复爆发时达到峰值,
然而,动作电位远高于单一动作,
潜力 这些动力学数据有助于解释这些机制
将电活动与基因表达相结合。 视网膜神经节细胞
(RGC)在活的鸡胚中可以用辣根标记
过氧化物酶。 这些RGC的轴突的生长锥是可视化的
并在不同的地方表现出戏剧性的变化,
视路(神经、视交叉和视束)。 这一准备工作应
允许对视觉的不同操作的效果进行评价
系统(电刺激、蛋白酶抑制剂、细胞粘附分子
抗体)对生长锥形态和轴突投射的影响
模式.
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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P G NELSON其他文献
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{{ truncateString('P G NELSON', 18)}}的其他基金
NEUROBIOLOGIC STUDIES OF NEURONS AND GLIA IN CELL CULTURE
细胞培养中神经元和神经胶质细胞的神经生物学研究
- 批准号:
3965719 - 财政年份:
- 资助金额:
-- - 项目类别:
NEUROBIOLOGIC STUDIES OF NEURONS AND GLIA IN CELL CULTURE
细胞培养中神经元和神经胶质细胞的神经生物学研究
- 批准号:
3842235 - 财政年份:
- 资助金额:
-- - 项目类别:
NEUROBIOLOGIC STUDIES OF NEURONS AND GLIA IN CELL CULTURE
细胞培养中神经元和神经胶质细胞的神经生物学研究
- 批准号:
3878029 - 财政年份:
- 资助金额:
-- - 项目类别:
NEUROBIOLOGIC STUDIES OF NEURONS AND GLIA IN CELL CULTURE
细胞培养中神经元和神经胶质细胞的神经生物学研究
- 批准号:
3898461 - 财政年份:
- 资助金额:
-- - 项目类别:
NEUROBIOLOGIC STUDIES OF NEURONS AND GLIA IN CELL CULTURE
细胞培养中神经元和神经胶质细胞的神经生物学研究
- 批准号:
4693707 - 财政年份:
- 资助金额:
-- - 项目类别:
NEUROBIOLOGIC STUDIES OF NEURONS AND GLIA IN CELL CULTURE
细胞培养中神经元和神经胶质细胞的神经生物学研究
- 批准号:
6162399 - 财政年份:
- 资助金额:
-- - 项目类别:
NEUROBIOLOGIC STUDIES OF NEURONS AND GLIA IN CELL CULTURE
细胞培养中神经元和神经胶质细胞的神经生物学研究
- 批准号:
3942010 - 财政年份:
- 资助金额:
-- - 项目类别:
NEUROBIOLOGIC STUDIES OF NEURONS AND GLIA IN CELL CULTURE
细胞培养中神经元和神经胶质细胞的神经生物学研究
- 批准号:
3756616 - 财政年份:
- 资助金额:
-- - 项目类别:
NEUROBIOLOGIC STUDIES OF NEURONS AND GLIA IN CELL CULTURE
细胞培养中神经元和神经胶质细胞的神经生物学研究
- 批准号:
3857046 - 财政年份:
- 资助金额:
-- - 项目类别:
NEUROBIOLOGIC STUDIES OF NEURONS AND GLIA IN CELL CULTURE
细胞培养中神经元和神经胶质细胞的神经生物学研究
- 批准号:
5203272 - 财政年份:
- 资助金额:
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