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.

项目摘要/摘要 受感受器死亡引起的视力丧失是不可逆转的失明的主要原因 在全球范围内，热选择仍然有限。因此，NEI的视网膜疾病计划已有 确定制定将残留声明作为核心计划目标的战略。 最近，一些实验室从干细胞中衍生了感光体，使细胞替代 疗法特别有希望。此外，对操纵的重要进展 可以刺激视网膜müller神经胶质的残留再生。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电路中，以重新建立其 功能。但是，我们仍然对基于正常布线的机制完全了解 感光体进入视网膜电路，尤其是用于锥形光感受器。 不同亚型的锥形光感受器将其连接到特定的圆圈中，因此功能性 可以探索差异（例如光谱灵敏度），以从 视觉场景。我们对该提案的主要假设是，每个锥形亚型都表达了特定的基因，这些基因是这些基因 允许其突触后伴侣（双极和水平细胞）认识，我们的主要目标是识别 这些基因。为了实现这一目标，我们将生成四个的完整转录组填充 斑马鱼中的不同锥形亚型，并鉴定出不同表达的基因（AIM＃1）。基于 这种差异表达，我们将执行反向遗传屏幕，我们将评估功能， 锥突触的结构和超微结构完整性（AIM＃2）。这将使我们能够确定 控制控制锥与其他永久细胞之间突触的形成，并促进 将锥体整合到视网膜电路中。我们认为，这种新知识可以直接应用 在改善视网膜变性的细胞替代或再生疗法中。而且，布线 通常，特定的孔是神经回路的关键特征。该提案从实验中有益 视网膜的可及性以及我们对视网膜细胞类型和电路的深刻了解，但是我们的方法具有 影响其他神经元退行性疾病的研究的潜力。