Masek Markus, Zang Jingjing, Mateos José M, Garbelli Marco, Ziegler Urs, Neuhauss Stephan C F, Bachmann-Gagescu Ruxandra
Institute of Medical Genetics, University of Zurich, Zurich, Switzerland; Department of Molecular Life Sciences, University of Zurich, Zurich, Switzerland.
Department of Molecular Life Sciences, University of Zurich, Zurich, Switzerland.
Methods Cell Biol. 2023;175:97-128. doi: 10.1016/bs.mcb.2022.10.004. Epub 2022 Dec 13.
Vision is one of our dominant senses and its loss has a profound impact on the life quality of affected individuals. Highly specialized neurons in the retina called photoreceptors convert photons into neuronal responses. This conversion of photons is mediated by light sensitive opsin proteins, which are found in the outer segments of the photoreceptors. These outer segments are highly specialized primary cilia, explaining why retinal dystrophy is a key feature of ciliopathies, a group of diseases resulting from abnormal and dysfunctional cilia. Therefore, research on ciliopathies often includes the analysis of the retina with special focus on the photoreceptor and its outer segment. In the last decade, the zebrafish has emerged as an excellent model organism to study human diseases, in particular with respect to the retina. The cone-rich retina of zebrafish resembles the fovea of the human macula and thus represents an excellent model to study human retinal diseases. Here we give detailed guidance on how to analyze the morphological and ultra-structural integrity of photoreceptors in the zebrafish using various histological and imaging techniques. We further describe how to conduct functional analysis of the retina by electroretinography and how to prepare isolated outer segment fractions for different -omic approaches. These different methods allow a comprehensive analysis of photoreceptors, helping to enhance our understanding of the molecular and structural basis of ciliary function in health and of the consequences of its dysfunction in disease.
视觉是我们主要的感官之一，视力丧失会对受影响个体的生活质量产生深远影响。视网膜中被称为光感受器的高度特化神经元将光子转化为神经反应。光子的这种转化由光敏感视蛋白介导，视蛋白存在于光感受器的外段。这些外段是高度特化的初级纤毛，这就解释了为什么视网膜营养不良是纤毛病的一个关键特征，纤毛病是一组由异常和功能失调的纤毛引起的疾病。因此，对纤毛病的研究通常包括对视网膜的分析，特别关注光感受器及其外段。在过去十年中，斑马鱼已成为研究人类疾病的优秀模式生物，尤其是在视网膜方面。斑马鱼富含视锥细胞的视网膜类似于人类黄斑的中央凹，因此是研究人类视网膜疾病的优秀模型。在这里，我们提供了关于如何使用各种组织学和成像技术分析斑马鱼光感受器的形态和超微结构完整性的详细指南。我们还描述了如何通过视网膜电图对视网膜进行功能分析，以及如何为不同的组学方法制备分离的外段组分。这些不同的方法能够对光感受器进行全面分析，有助于增进我们对健康状态下纤毛功能的分子和结构基础以及疾病中纤毛功能障碍后果的理解。
