Retinal circuit disassembly in primate glaucoma

项目摘要/摘要 在发育过程中，特定的突触伙伴相互连接，以确保适当的神经回路功能， 但这些连接在神经退化过程中是如何分解的，人们还不太清楚。在啮齿动物中 实验性青光眼(EG)模型，突触丢失发生在视网膜神经节细胞(RGC)树突之前 退缩和细胞死亡。在啮齿动物身上的聚合证据表明，特定类型的RGC更容易受到影响 与眼压升高有关，但对青光眼灵长类动物视网膜环路解体知之甚少 视网膜。事实上，在缺乏筛板和黄斑并具有不同RGC的小鼠中，存在显著的差异 类型，限制了对人类研究结果的翻译和推广。解决这一问题至关重要 知识鸿沟，以促进临床相关诊断和新疗法的成功开发 方法，如神经保护、基因治疗和基于细胞的视力恢复策略。在这里我们 组建一支高度协作的调查团队，与我们的目标完全匹配，并具有互补的专业知识 系统地确定RGC的连接性、功能和转录本 以及青光眼灵长类动物视网膜中突触的分解。我们的方法建立在一只久负盛名的恒河猴身上 猕猴非人类灵长类(NHP)实验性青光眼模型 在人类青光眼中观察到的功能变化，可以对疾病进行详细和准确的分期。基座 根据我们在小鼠身上的研究和NHP的初步数据，我们假设在受伤的人中有特定的微电路 成人NHP视网膜可能在连通性和功能上表现出易感性，这反映在差异 基因表达。为了验证这一假设，我们使用了严格的定量电生理学、解剖学和 NHP视网膜中四种主要RGC类型的分子评估：开启和关闭侏儒和遮阳伞 神经节细胞。AIM 1将使用高密度多电极阵列、单电池记录和Patch-Seq来识别 NHP EG易感的功能性RGC类型及其转录本的研究 洞察力和新的治疗目标。目标2将确定回路和突触的特异性和模式 在NHP EG中，从板层特定和细胞类型特定的角度使用详细的电路和 突触映射。该方案具有创新性，因为它结合了多峰函数、形态和 分子分析，具有重要意义，因为它集中在灵长类动物中四种主要的RGC类型，这四种类型解释了 人类的大部分视力都受到青光眼的影响。我们将为科学研究创造大量资源 社区，并揭示了对视网膜电路解体和电路修复的潜力的洞察 临床相关的青光眼模型。

PROJECT SUMMARY / ABSTRACT During development, specific synaptic partners connect in order to ensure proper neural circuit function, but how these connections are disassembled during neurodegeneration is less well understood. In rodent experimental glaucoma (EG) models, synapse loss occurs early, preceding retinal ganglion cell (RGC) dendrite retraction and cell death. Converging evidence in rodents suggests that specific RGC types are more susceptible to elevated intraocular pressure, but little is known about retinal circuit disassembly in glaucomatous primate retina. Indeed, significant differences in mice, which lack a lamina cribrosa and macula and have dissimilar RGC types, limit the translation and generalizability of findings to humans. It is critically important to address this knowledge gap in order to advance successful development of clinically relevant diagnostics and novel treatment approaches, such as neuroprotection, gene therapy, and cell-based vision restoration strategies. Here we assemble a highly collaborative team of investigators with complementary expertise well-matched to our goal of systematically determining the connectivity, function, and transcriptomes of RGCs undergoing circuit and synapse disassembly in glaucomatous primate retina. Our approach builds on a well-established rhesus macaque non-human primate (NHP) model of experimental glaucoma that closely recapitulates structural and functional changes observed in human glaucoma, and permits detailed and precise staging of disease. Based on our studies in mice and preliminary data in NHP, we hypothesize that specific microcircuits in the injured adult NHP retina may exhibit susceptibility in connectivity and function, which is reflected in differential gene expression. To test this hypothesis, we apply rigorous quantitative electrophysiological, anatomical, and molecular assessments focusing on the four main RGC types in NHP retina: ON and OFF midget and parasol ganglion cells. Aim 1 will use high-density multielectrode arrays, single cell recordings, and Patch-seq to identify the functional RGC types that are vulnerable in NHP EG and probe their transcriptomes to reveal mechanistic insights and novel therapeutic targets. Aim 2 will determine the specificity and patterns of circuit and synapse disassembly in NHP EG from both lamina-specific and cell type-specific perspectives using detailed circuit and synapse mapping. The proposal is innovative because it brings together multi-modal function, morphologic, and molecular analyses, and is significant because it focuses on the four main RGC types in primate that account for the majority of human vision and are affected in glaucoma. We will generate significant resources for the scientific community and reveal insights into retinal circuit disassembly and the potential for circuit repair in a highly clinically relevant model of glaucoma.