Li Qihang, Zhu He, Fan Miaomiao, Sun Jing, Reinach Peter S, Wang Yuhan, Qu Jia, Zhou Xiangtian, Zhao Fuxin
School of Optometry and Ophthalmology and Eye Hospital, Wenzhou Medical University, Wenzhou, Zhejiang, China; State Key Laboratory of Optometry, Ophthalmology and Vision Science, Wenzhou, Zhejiang, China.
School of Optometry and Ophthalmology and Eye Hospital, Wenzhou Medical University, Wenzhou, Zhejiang, China; State Key Laboratory of Optometry, Ophthalmology and Vision Science, Wenzhou, Zhejiang, China; Research Unit of Myopia Basic Research and Clinical Prevention and Control, Chinese Academy of Medical Sciences (2019RU025), Wenzhou, Zhejiang, China; Oujiang Laboratory, Zhejiang Lab for Regenerative Medicine, Vision and Brain Health, Wenzhou, Zhejiang, China.
Exp Eye Res. 2022 May;218:109018. doi: 10.1016/j.exer.2022.109018. Epub 2022 Mar 1.
The process of eye axis lengthening in myopic eyes is regulated by multiple mechanisms in the retina, and horizontal cells (HCs) are an essential interneuron in the visual regulatory system. Wherein intracellular Ca plays an important role in the events involved in the regulatory role of HCs in the retinal neural network. It is unknown if intracellular Ca regulation in HCs mediates changes in the retinal neural network during myopia progression. We describe here a novel calcium fluorescence indicator system that monitors HCs' intracellular Ca levels during form-deprivation myopia (FDM) in mice. AAV injection of GCaMP6s, as a protein calcium sensor, into a Gja10-Cre mouse monitored the changes in Casignaling in HC that accompany FDM progression in mice. An alternative Gja10-Cre/Ai96-GCaMP6s mouse model was created by cross mating Gja10-Cre with Ai96 mice. Immunofluorescence imaging and live imaging of the retinal cells verified the identity of these animal models. Changes in retinal horizontal cellular Ca levels were resolved during FDM development. The numbers of GCaMP6s and the proportion of HCs were tracked based on profiling changes in GCaMP6scalbindin/calbindin coimmunostaining patterns. They significantly decreased more after either two days (P < 0.01) or two weeks (P < 0.001) in form deprived eyes than in the untreated fellow eyes. These decreases in their proportion reached significance only in the retinal central region rather than also in the retinal periphery. A novel approach employing a GCaMP6s mouse model was developed that may ultimately clarify if HCs mediate Ca signals that contribute to controlling FDM progression in mice. The results indicate so far that FDM progression is associated with declines in HC Ca signaling activity.
近视眼中眼轴延长的过程受视网膜中多种机制调控，而水平细胞（HCs）是视觉调节系统中一种重要的中间神经元。其中细胞内钙离子在水平细胞对视网膜神经网络的调节作用所涉及的事件中发挥重要作用。目前尚不清楚水平细胞内钙离子调节是否介导近视进展过程中视网膜神经网络的变化。我们在此描述一种新型钙荧光指示剂系统，该系统可监测小鼠形觉剥夺性近视（FDM）过程中水平细胞的细胞内钙离子水平。通过向Gja10-Cre小鼠注射作为蛋白质钙传感器的GCaMP6s，监测了小鼠FDM进展过程中水平细胞内钙信号的变化。通过将Gja10-Cre与Ai96小鼠杂交，创建了另一种Gja10-Cre/Ai96-GCaMP6s小鼠模型。视网膜细胞的免疫荧光成像和实时成像验证了这些动物模型的特性。在FDM发展过程中解析了视网膜水平细胞钙水平的变化。基于GCaMP6s钙结合蛋白/钙结合蛋白共免疫染色模式的分析变化，追踪了GCaMP6s的数量和水平细胞的比例。与未处理的对侧眼相比，形觉剥夺眼在两天（P < 0.01）或两周（P < 0.001）后，它们的比例显著下降得更多。它们比例的这些下降仅在视网膜中央区域达到显著水平，而非在视网膜周边区域。开发了一种采用GCaMP6s小鼠模型的新方法，该方法最终可能阐明水平细胞是否介导有助于控制小鼠FDM进展的钙信号。目前的结果表明，FDM进展与水平细胞钙信号活性的下降有关。
