Identification of genes involved in photoreceptor recognition and synapse formation

鉴定参与光感受器识别和突触形成的基因

• 批准号: 10766366
• 负责人: Juan Angueyra
• 金额: $ 24.9万
• 依托单位: UNIV OF MARYLAND, COLLEGE PARK
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-01 至 2026-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10766366
• 关键词: Address; Adult; Award; Blindness; CRISPR/Cas technology; Candidate Disease Gene; Cause of Death; Cells; Color; Cone; Degenerative Disorder; Developed Countries; Development; Dissociation; Electron Microscopy; Electroretinography; Exhibits; Gene Expression Profile; Genes; Genetic Screening; Goals; Immunohistochemistry; Institution; Knowledge; Label; Laboratories; Larva; Measures; Morphology; Muller' s cell; Mutagenesis; National Eye Institute; Nerve Degeneration; Nervous System; Neurons; Opsin; Photoreceptors; Principal Investigator; Process; Public Health; Replacement Therapy; Reporter; Repression; Research; Retina; Retinal Cone; Retinal Degeneration; Retinal Diseases; Sorting; Specificity; Structure; Synapses; Testing; Therapeutic; Transgenic Organisms; Transplantation; Vision; Visual; Zebrafish; aspirate; career; cell replacement therapy; cell type; differential expression; direct application; disability; genetic profiling; horizontal cell; improved; insight; knockout gene; molecular recognition; mutant; neural circuit; novel; postsynaptic; programs; regenerative; regenerative therapy; retinal regeneration; retinal stimulation; reverse genetics; ribbon synapse; stem cells; success; synaptogenesis; therapy development; tool; transcriptome sequencing; transcriptomic profiling; treatment strategy

PROJECT SUMMARY/ABSTRACT Vision loss caused by the death of photoreceptors is a leading cause of irreversible blindness worldwide, yet therapeutic options remain limited. For this reason, the NEI's Retinal Disease Program has identified the development of strategies for the treatment of retinal degenerations as a core program goal. Recently, several laboratories have derived photoreceptors from stem cells, making cell-replacement therapies particularly promising. Additionally, important advances have been made into manipulations that could stimulate retinal regeneration from the retinal Müller glia. The critical barrier for the success of such therapies is the integration of derived photoreceptors into existing retinal circuits to reestablish their function. Yet, we still lack a complete understanding on the mechanisms that underlie the normal wiring of photoreceptors into retinal circuits, especially for cone photoreceptors. Cone photoreceptors of different subtypes are wired into specific retinal circuits, so that functional differences (like spectral sensitivity) may be exploited to extract specific information (like color) from the visual scene. Our main hypothesis for this proposal is that each cone subtype expresses specific genes that allow recognition by its postsynaptic partners (bipolar and horizontal cells), and our main goal is to identify these genes. To accomplish this, we will first generate a complete transcriptomic profiling of the four different cone subtypes in zebrafish, and identify genes that are differentially expressed (aim #1). Based on this differential expression, we will perform a reverse-genetic screen, where we will assess the functional, structural and ultrastructural integrity of the cone synapses (aim #2). This will allow us to identify genes that control the control the formation of synapses between cones and other retinal cells, and that promote the integration of cones into retinal circuits. We believe that this new knowledge could have direct applications in the improvement of cell-replacement or regenerative therapies for retinal degenerations. Moreover, wiring specificity is a key feature of neural circuits in general. This proposal benefits from the experimental accessibility of the retina and our deep knowledge of retinal cell types and circuits, but our approach has the potential to impact the study of other neuronal degenerative diseases.

项目概要/摘要 光感受器死亡引起的视力丧失是不可逆失明的主要原因 在世界范围内，但治疗选择仍然有限，因此 NEI 的视网膜疾病计划已制定。 将制定治疗视网膜变性的策略确定为核心计划目标。 最近，一些实验室从干细胞中提取出光感受器，使细胞替代成为可能。 此外，治疗方法也取得了重要进展，这些进展特别有希望。 可以刺激视网膜穆勒神经胶质细胞的再生，这是这种成功的关键障碍。 疗法是将衍生的光感受器整合到现有的视网膜回路中，以重建它们的 然而，我们仍然对正常接线的机制缺乏完整的了解。 感光器进入视网膜回路，特别是锥体感光器。 不同亚型的视锥细胞被连接到特定的视网膜电路中，因此功能 可以利用差异（如光谱灵敏度）从图像中提取特定信息（如颜色） 我们对这个提议的主要假设是每个视锥细胞亚型都表达特定的基因。 允许其突触后伙伴（双极和水平细胞）识别，我们的主要目标是识别 为了实现这一目标，我们将首先生成这四个基因的完整转录组分析。 斑马鱼的不同锥体亚型，并识别差异表达的基因（目标＃1）。 这种差异表达，我们将进行反向遗传筛选，我们将评估功能， 锥体突触的结构和超微结构完整性（目标＃2）这将使我们能够识别基因。 控制视锥细胞和其他视网膜细胞之间突触的形成，并促进 我们相信这一新知识可以直接应用。 改善视网膜变性的细胞替代或再生疗法。 一般来说，特异性是神经回路的一个关键特征。该提议受益于实验。 视网膜的可及性以及我们对视网膜细胞类型和电路的深入了解，但我们的方法具有 潜在影响其他神经元退行性疾病的研究。