Department of Biomedical Engineering, University of Rochester, Rochester, NY, 14620, USA.
Center for Visual Science, University of Rochester, Rochester, NY, 14627, USA.
Commun Biol. 2023 May 29;6(1):572. doi: 10.1038/s42003-023-04896-x.
The laboratory mouse has provided tremendous insight to the underpinnings of mammalian central nervous system physiology. In recent years, it has become possible to image single neurons, glia and vascular cells in vivo by using head-fixed preparations combined with cranial windows to study local networks of activity in the living brain. Such approaches have also succeeded without the use of general anesthesia providing insights to the natural behaviors of the central nervous system. However, the same has not yet been developed for the eye, which is constantly in motion. Here we characterize a novel head-fixed preparation that enables high-resolution adaptive optics retinal imaging at the single-cell level in awake-behaving mice. We reveal three new functional attributes of the normal eye that are overlooked by anesthesia: 1) High-frequency, low-amplitude eye motion of the mouse that is only present in the awake state 2) Single-cell blood flow in the mouse retina is reduced under anesthesia and 3) Mouse retinae thicken in response to ketamine/xylazine anesthesia. Here we show key benefits of the awake-behaving preparation that enables study of retinal physiology without anesthesia to study the normal retinal physiology in the mouse.
实验室小鼠为哺乳动物中枢神经系统生理学的基础提供了巨大的洞察力。近年来，通过使用头部固定装置结合颅窗，可以对单个神经元、神经胶质细胞和血管细胞进行体内成像，从而研究活体大脑中局部活动网络。这些方法也成功地应用于不使用全身麻醉的情况下，提供了对中枢神经系统自然行为的洞察。然而，对于眼睛这一不断运动的器官，还没有开发出同样的方法。在这里，我们描述了一种新的头部固定装置，该装置能够在清醒行为的小鼠中实现单细胞水平的高分辨率自适应光学视网膜成像。我们揭示了正常眼睛的三个新的功能属性，这些属性被麻醉所忽视：1）仅在清醒状态下出现的小鼠高频、低幅度的眼球运动；2）麻醉状态下小鼠视网膜的单个细胞血流减少；3）小鼠视网膜在氯胺酮/甲苯噻嗪麻醉下增厚。在这里，我们展示了清醒行为准备的关键优势，它能够在不使用麻醉的情况下研究视网膜生理学，从而研究小鼠的正常视网膜生理学。
