VIDEO MICROSCOPY OF FAST AXONAL-TRANSPORT IN EXTRUDED AXOPLASM - A NEW MODEL FOR STUDY OF MOLECULAR MECHANISMS
VIDEO MICROSCOPY OF FAST AXONAL-TRANSPORT IN EXTRUDED AXOPLASM - A NEW MODEL FOR STUDY OF MOLECULAR MECHANISMS
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DOI：
10.1002/cm.970050203
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发表时间：
1985-01-01
影响因子：
--
通讯作者：
ALLEN, RD
中科院分区：
文献类型：
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作者：
BRADY, ST;LASEK, RJ;ALLEN, RD
AVEC-DIC（艾伦视频增强衬度，微分干涉衬度）显微镜的发展以及将该方法应用于研究从乌贼Loligo pealei巨轴突中挤出的分离轴浆中的快速轴突运输，为分析膜细胞器的细胞内运输提供了新的范例。轴突的位置、传输颗粒的数量以及该制剂中不存在像质膜这样的渗透性屏障，允许进行许多使用其他模型系统难以或不可能进行的实验。详细描述了该制剂的用途和特点，并首次评估了许多性能。挤压过程的特征在于。颗粒运动在挤压轴浆的内部和沿着从灌注轴浆外围延伸的单个原纤维进行评估。分析了二价阳离子，特别是 Ca2+ 的作用，以及 Ca2+ 升高对轴浆组织和运输的影响。使用一系列改变细胞骨架组织的药理学试剂和多肽来检查微丝和微管在快速运输中的作用。讨论了消耗 ATP 和添加 ATP 类似物的影响。挤出的轴浆制剂被证明是细胞内运输分子机制的生化和药理学分析的宝贵模型系统。
The development of AVEC-DIC [Allen video-enhanced contrast, differential interference contrast] microscopy and the application of this method to the study of fast axonal transport in isolated axoplasm extruded from the giant axon of the squid Loligo pealei provides a new paradigm for analyzing the intracellular transport of membranous organelles. The site of the axon, the number of transported particles, and the absence of permeability barriers like the plasma membrane in this preparation permit many experiments that are difficult or impossible to perform using other model systems. The use and features of this preparation are described in detail and a number of properties are evaluated for the 1st time. The process of extrusion is characterized. Particle movement is evaluated both in the interior of extruded axoplasm and along individual fibrils that extend from the periphery of perfused axoplasm. The role of divalent cations, particularly Ca2+, and the effects of elevated Ca2+ on axoplasmic organization and transport are analyzed. A series of pharmacological agents and polypeptides that alter cytoskeletal organization are used to examine the role of microfilaments and microtubules in fast transport. The effects of depleting ATP and of adding ATP analogs are discussed. The extruded axoplasm preparation is shown to be an invaluable model system for biochemical and pharmacological analyses of the molecular mechanisms of intracellular transport.