Sichuan Provincial Key Laboratory for Human Disease Gene Study, Sichuan Provincial People's Hospital, University of Electronic Science and Technology of China.
Sichuan Provincial Key Laboratory for Human Disease Gene Study, Sichuan Provincial People's Hospital, University of Electronic Science and Technology of China; School of Medicine, University of Electronic Science and Technology of China.
J Vis Exp. 2024 Oct 4(212). doi: 10.3791/66991.
Glaucoma is a leading cause of blindness globally, characterized by a complex pathogenic mechanism that makes vision restoration challenging. Mice serve as valuable animal models for studying the pathogenesis and treatment of glaucoma due to their relatively homogenous genetic background and retinal ganglion cells (RGCs), which structurally resemble those in humans. Accurate assessment of RGC damage and treatment outcomes in mouse glaucoma models requires determining the RGC number across the entire retina. This protocol outlines a comprehensive method involving the isolation of the whole retina, labeling RGCs with specific antibodies, and rapid, accurate automatic counting of RGCs using an AI-based program. The streamlined approach allows for efficient and precise quantification of RGC numbers in mouse retinas, facilitating the evaluation of RGC degeneration and potential therapeutic interventions. By enabling researchers to assess the extent of RGC damage, this protocol contributes to a deeper understanding of glaucoma pathogenesis and aids in developing effective treatment strategies to manage and prevent vision loss.
青光眼是全球范围内导致失明的主要原因之一，其发病机制复杂，使得视力恢复具有挑战性。由于其相对同质的遗传背景和类似于人类的视网膜神经节细胞（RGCs），小鼠成为研究青光眼发病机制和治疗方法的有价值的动物模型。要准确评估小鼠青光眼模型中的 RGC 损伤和治疗效果，需要确定整个视网膜中的 RGC 数量。本方案概述了一种全面的方法，包括全视网膜分离、用特定抗体标记 RGC 以及使用基于人工智能的程序快速、准确地自动计数 RGC。这种简化的方法可以高效、精确地定量小鼠视网膜中的 RGC 数量，有助于评估 RGC 退化和潜在的治疗干预效果。通过使研究人员能够评估 RGC 损伤的程度，本方案有助于深入了解青光眼的发病机制，并有助于开发有效的治疗策略来管理和预防视力丧失。
