MOTILE MODELS OF TELEOST RETINAL CONES

TELEOST 视网膜锥体的运动模型

基本信息

批准号：
3281533
负责人：
MARY BETH BURNSIDE
金额：
$ 8.72万
依托单位：
UNIVERSITY OF CALIFORNIA BERKELEY
依托单位国家：
美国
项目类别：
财政年份：
1983
资助国家：
美国
起止时间：
1983-08-01 至 1986-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/3281533
关键词：
aquatic organism biological models calcium channel blockers cell motility cyclic AMP electron microscopy immunofluorescence technique monoclonal antibody nucleotides smooth muscle tissue /cell culture

项目摘要

The general objective of this research program is to use lysed-cell models of teleost retinal cones to investigate the mechanisms and regulation of actomyosin-dependent cell contraction and microtubule-dependent cell elongation. We have developed procedures for obtaining ATP-dependent re-activation of both contraction and elongation in models of teleost cones lysed with 1% Brij-58. By altering cAMP and calcium levels we can produce either reactivated elongation or contraction with sufficiently extensive length changes (20-40u) to permit detailed quantitation. We will analyze the effects of reactivation conditions and various probes by quantifying rates and extents of reactivated movement and by thin section TEM of models. We propose specifically 1) to examine the roles of myosin, myosin light chain kinase and calmodulin in regulating cone contraction and 2) to investigate the possibility that dynein-based inter-microtubule sliding provides motive force for cone elongation. Proposed studies include analyses of the effects of nucleotides, ions, pH, vanadate and EHNA on reactivated elongation and contraction, immunofluorescence localization and ferritin-antibody localization studies with antibodies to platelet myosin, calmodulin, and ciliary and egg dynein, investigations of the effects of exogenous calmodulin, smooth muscle myosin and myosin light chain kinase on reactivated contraction, and morphological analyses of microtubule polarities and intertubule associations in elongating cone myoids.

该研究计划的总体目标是使用裂解细胞模型硬骨鱼视网膜锥体的研究机制和调节肌动球蛋白依赖性细胞收缩和微管依赖性细胞伸长。我们已经开发了获得 ATP 依赖性的程序硬骨鱼锥体模型中收缩和伸长的重新激活用 1% Brij-58 裂解。通过改变 cAMP 和钙水平，我们可以产生要么重新激活伸长，要么收缩足够广泛长度变化 (20-40u) 以允许详细定量。我们将分析通过量化重新激活条件和各种探针的影响重新激活运动的速率和程度以及通过薄切片 TEM 模型。我们特别建议1）检查肌球蛋白的作用，肌球蛋白轻链激酶和钙调蛋白调节视锥细胞收缩，2) 研究基于动力蛋白的微管间滑动的可能性为锥体伸长提供动力。拟议的研究包括分析核苷酸、离子、pH、钒酸盐和 EHNA 对重新激活伸长和收缩，免疫荧光定位和使用血小板肌球蛋白抗体进行铁蛋白抗体定位研究，钙调蛋白、纤毛和卵动力蛋白，影响的研究外源性钙调蛋白、平滑肌肌球蛋白和肌球蛋白轻链激酶重新激活的收缩和微管的形态分析延长锥体肌样中的极性和小管间关联。