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Towards the vertebrate eye - Exploring photoreceptors of basal deuterostomes

走向脊椎动物的眼睛 - 探索基底后口动物的光感受器

基本信息

批准号：
251841938
负责人：
Professor Dr. Carsten Lüter, since 3/2018
金额：
--
依托单位：
Museum für Naturkunde - Leibniz-Institut für Evolutions- und Biodiversitätsforschung (MfN)
依托单位国家：
德国
项目类别：
Research Grants
财政年份：
2014
资助国家：
德国
起止时间：
2013-12-31 至 2019-12-31
项目状态：
已结题

来源：
https://gepris.dfg.de/gepris/projekt/251841938?language=en
关键词：
Towards vertebrate eye Exploring photoreceptors

项目摘要

The evolution of eye organs is a matter of scientific debate since more than 150 years. Investigating light sensitive organs and photoreceptor cell (PRC) types in Ambulacraria (echinoderms + enteropneusts) - the most basal recent deuterostome phyla - will elucidate the evolution of specific PRC types and systems eventually leading to the development of the vertebrate eye. This will be achieved by deployment of an integrative approach with a broad set of molecular, structural, behavioural and functional molecular methods. Antibody stainings against various opsin photopigments will allow identification of hitherto unknown PRCs in a broad taxon sampling of echinoderm and enteropneust species. Subsequent gene expression profiling (´molecular fingerprinting´) of the identified PRC types via in situ-hybridization will assist in elucidating proposed mechanisms driving (photoreceptor) cell type evolution (e.g. segregation of multifunctional cell types into sister cell types with reduced functions, independend gene co-option). The connection of the PRCs to the nervous system (and thus the mode of stimulus integration) is essential for our understanding of PRC function and will be investigated with immunohistochemical methods as well as with transmission electron microscopy. As different echinoderm subgroups have been demonstrated to deploy skeletal elements either as a shielding- or a focussing device, the functional role of echinoderm skeletal components will be studied by non-invasive X-ray microtomography in the original tissue context. The integrative dataset obtained will allow for phylogenetic comparison between ambulacrarian PRCs and those of other (deuterostome) phyla. Furthermore these data comprise the basis for the biophysical modelling of the optical properties of echinoderm and enteropneust PRC systems which is under construction in collaboration with Dan-E. Nilsson from The Vision Group of Lund University, Sweden. Functional molecular analysis, focussed on two different opsin proteins of the purple sea urchin Strongylocentrotus purpuratus will finally allow us to gather first direct evidence for the function of an echinoderm PRC system. Those experiments will be performed in close collaboration with Maria I. Arnone from Stazione Zoologica in Naples, and will deploy sea urchin 'conventional' gene knock down via morpholino injection as well gene knock out via Sigma customized Zinc Finger Technology. The summary of obtained data will allow us to draw important conclusions about the evolution of PRCs at the base of deuterostomes as well as about PRC properties functioning in an evolutionary 'ancient' marine ecological context.

眼睛器官的进化是150多年来科学争论的问题。调查光敏感器官和感光细胞（PRC）类型Ambulacraria（棘皮动物+enterocentrist）-最近最基本的后口门-将阐明特定的PRC类型和系统的进化，最终导致脊椎动物眼睛的发展。这将通过部署具有广泛的分子、结构、行为和功能分子方法的综合方法来实现。针对各种视蛋白色素的抗体染色将允许在棘皮动物和肠栖动物物种的广泛分类群采样中识别迄今未知的PRCs。随后通过原位杂交鉴定的PRC类型的基因表达谱（“分子指纹”）将有助于阐明驱动（光感受器）细胞类型进化的拟议机制（例如，多功能细胞类型分离为功能降低的姐妹细胞类型，独立基因共选择）。PRC与神经系统的连接（以及刺激整合的模式）对于我们理解PRC的功能至关重要，将用免疫组织化学方法和透射电子显微镜进行研究。由于不同的棘皮动物亚组已被证明部署骨骼元素作为屏蔽或聚焦设备，棘皮动物骨骼组件的功能作用将通过非侵入性X射线显微断层扫描在原始组织背景下进行研究。所获得的综合数据集将允许步足类PRC和其他（后口类）门之间的系统发育比较。此外，这些数据构成了棘皮动物和肠栖动物PRC系统光学特性生物物理建模的基础，该系统正在与Dan-E合作建设中。Nilsson来自瑞典隆德大学视觉小组。功能分子分析，集中在两个不同的视蛋白蛋白质的紫海胆Strongylocentrotus purpuratus将最终使我们能够收集第一个直接的证据棘皮动物PRC系统的功能。这些实验将与玛丽亚一世密切合作进行。Arnone来自那不勒斯的Stazione Zoologica，并将通过吗啉注射部署海胆“常规”基因敲除，以及通过Sigma定制的锌指技术进行基因敲除。所获得的数据的摘要将使我们能够得出重要的结论，在后口动物的基础上，以及有关中国人民共和国的性能功能在进化的“古老”的海洋生态环境的演变。