In Vivo Function and Metabolism Evaluation of Glaucomatous RGCs by Two-Photon Scanning Laser Ophthalmology

双光子扫描激光眼科评价青光眼 RGC 的体内功能和代谢

• 批准号: 10660761
• 负责人: Yang Hu
• 金额: $ 64.15万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2028-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10660761
• 关键词: Acute; Anatomy; Angle-Closure Glaucoma; Animal Model; Animals; Belief; Biological Markers; Biosensor; Blindness; Blood Vessels; Cell physiology; Cells; Cessation of life; Characteristics; Chronic; Color; Combined Modality Therapy; Contact Lenses; Cornea; Data; Dehydration; Detection; Development; Disease; Disease Progression; Electrophysiology (science); Evaluation; Excision; Fluorescence; Functional Imaging; Functional disorder; Fundus photography; Glaucoma; Image; Imaging Techniques; Individual; Interdisciplinary Study; Investigation; Label; Lasers; Light; Lighting; Metabolic; Metabolism; Modeling; Monitor; Morphology; Motion; Mus; Nerve Degeneration; Neural Retina; Neurons; Ocular Hypertension; Open-Angle Glaucoma; Ophthalmology; Ophthalmoscopy; Optics; Pattern; Phenotype; Photic Stimulation; Photons; Photoreceptors; Physiologic Intraocular Pressure; Physiology; Predisposition; Property; Protocols documentation; Retina; Retinal Diseases; Retinal Ganglion Cells; Scanning; Severities; Silicone Oils; Spottings; System; Testing; Time; Transgenic Mice; Traumatic injury; Update; Visualization; calcium indicator; cell type; cellular imaging; clinically relevant; experimental study; imaging modality; imaging platform; imaging system; improved; in vivo; in vivo imaging; in vivo optical imaging; neural circuit; neuroprotection; non-invasive imaging; novel; overexpression; prevent; resilience; response; retinal imaging; retinal stimulation; sensor; two-photon; visual processing; visual stimulus

PROJECT SUMMARY Glaucoma, the most common worldwide cause of irreversible blindness, is characterized by progressive dysfunction and death of retinal ganglion cells (RGCs). We recently employed confocal scanning laser ophthalmoscopy (cSLO) to successfully obtain in vivo Ca2+ imaging with mouse RGCs expressing jGCaMP7s, a genetically encoded calcium indicator. Thousands of ON, OFF, and ON-OFF RGCs with characteristic responses to light stimulation are readily detected in living animals through this non-invasive in vivo imaging platform. Here we seek to develop a more advanced, first-of-its-kind two-photon (2P)-SLO platform with patterned stimulation and multiple detection channels. Through a multidisciplinary collaboration with expertise in in vivo optical imaging, RGC pathophysiology, and retinal neural circuitry and visual processing, we will use complementary imaging techniques and state-of-the-art analysis protocols to understand naïve RGC physiology in real time. We recently extended our original mouse silicone oil-induced pupillary blocking and ocular hypertension (SOHU) model to recapitulate phenotypes of two forms of glaucoma: a chronic model with moderate IOP elevation and mild RGC neurodegeneration; and an acute model with greatly elevated IOP and severe neurodegeneration. Importantly, SO removal reduces IOP to normal almost immediately, allowing better exploration of the effects of IOP lowering treatment and combined treatment with neuroprotection strategies. Thus, we will determine the longitudinal functional and metabolic changes of glaucomatous RGCs, under clinically relevant models, both before and after IOP normalization and/or neuroprotective treatments. These data will deepen our understanding of the pathophysiology of glaucoma, towards finding much-sought biomarkers to better predict progression, and create more relevant endpoints for developing treatment to restore RGC physiology in vivo.

项目总结 青光眼是世界范围内最常见的不可逆性失明原因，其特点是进行性 视网膜神经节细胞(RGC)功能障碍和死亡。我们最近使用了共聚焦扫描激光 眼底镜(CSLO)成功获得体内表达jGCaMP7，a的小鼠视网膜神经节细胞的钙成像 基因编码的钙指示剂。数千个具有特征响应的开、关和开-关RGC 通过这个非侵入性的活体成像平台，可以很容易地在活体动物中检测到光刺激。这里 我们寻求开发一种更先进的、同类中第一个具有图案化刺激的双光子(2P)-SLO平台 和多个检测通道。通过具有体内光学专业知识的多学科合作 成像，RGC病理生理学，以及视网膜神经电路和视觉处理，我们将使用互补 成像技术和最先进的分析协议，以实时了解幼稚的RGC生理。我们 最近延长了我们最初的小鼠硅油诱导的瞳孔阻塞和高眼压(搜狐) 总结两种类型青光眼表型的模型：中度眼压升高的慢性模型和 轻度RGC神经变性；急性模型眼压显著升高和严重神经变性。 重要的是，摘除SO几乎可以立即将眼压降至正常，从而可以更好地探索 降眼压治疗与神经保护策略相结合。因此，我们将确定 临床相关模型下青光眼视网膜节细胞的纵向功能和代谢变化 在眼压正常化和/或神经保护治疗前后。这些数据将加深我们对 青光眼的病理生理学，寻找人们期待已久的生物标记物来更好地预测病情进展，以及 为发展治疗创造更多相关的终点，以恢复RGC的活体生理。