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通过同时测量视觉色素荧光和 ERG,研究正常和减少 arrestin 含量的果蝇光感受器反应。

Photoreceptor responses of fruitflies with normal and reduced arrestin content studied by simultaneous measurements of visual pigment fluorescence and ERG.

机构信息

Department of Biology, Biotechnical Faculty, University of Ljubljana, 1000 Ljubljana, Slovenia.

出版信息

J Comp Physiol A Neuroethol Sens Neural Behav Physiol. 2010 Jan;196(1):23-35. doi: 10.1007/s00359-009-0489-5. Epub 2009 Nov 19.

Abstract

We have simultaneously measured the electroretinogram (ERG) and the metarhodopsin content via fluorescence in white-eyed, wild-type Drosophila and the arrestin2 hypomorphic mutant (w(-);arr2 (3)) at a range of stimulus wavelengths and intensities. Photoreceptor response amplitude and termination (transition between full repolarization and prolonged depolarizing afterpotential, PDA) were related to visual pigment conversions and arrestin concentration. The data were implemented in a kinetic model of the rhodopsin-arrestin cycle, allowing us to estimate the active metarhodopsin concentration as a function of effective light intensity and arrestin concentration. Arrestin reduction in the mutant modestly increased the light sensitivity and decreased the photoreceptor dynamic range. Compared to the wild type, in the mutant the transition between full repolarization and PDA occurred at a lower metarhodopsin fraction and was more abrupt. We developed a steady-state stochastic model to interpret the dependence of the PDA on effective light intensity and arrestin content and to help deduce the arrestin to rhodopsin ratio from the sensitivity and PDA data. The feasibility of different experimental methods for the estimation of arrestin content from ERG and PDA is discussed.

摘要

我们同时测量了白眼、野生型果蝇和 arrestin2 弱突变体(w(-);arr2 (3))的视网膜电图(ERG)和视黄醛含量的荧光,在一系列刺激波长和强度下。光感受器反应幅度和终止(完全复极化和延长去极化后电位之间的过渡,PDA)与视觉色素转化和 arrestin 浓度有关。这些数据被应用于视黄醛-arrestin 循环的动力学模型中,使我们能够估计有效光强度和 arrestin 浓度作为活性视黄醛浓度的函数。突变体中 arrestin 的减少适度增加了光敏感性并降低了光感受器的动态范围。与野生型相比,在突变体中,完全复极化和 PDA 之间的过渡发生在较低的视黄醛分数处,并且更加突然。我们开发了一个稳态随机模型来解释 PDA 对有效光强度和 arrestin 含量的依赖性,并帮助从敏感性和 PDA 数据中推断出 arrestin 与视黄醛的比例。讨论了从 ERG 和 PDA 估计 arrestin 含量的不同实验方法的可行性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a737/2797847/93f029bea39a/359_2009_489_Fig1_HTML.jpg

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