Axonal architecture of the mouse inner retina revealed by second harmonic generation.
Axonal architecture of the mouse inner retina revealed by second harmonic generation.
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小鼠内部视网膜的轴突结构由第二次谐波产生揭示。
DOI：
10.1093/pnasnexus/pgac160
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发表时间：
2022-09
期刊：
影响因子：
--
通讯作者：
Lim, Hyungsik
中科院分区：
文献类型：
--
作者：
Meah, Arafat;Boodram, Vinessia;Bucinca-Cupallari, Festa;Lim, Hyungsik
我们描述了一种新的方法来可视化未标记的新鲜完整视网膜中的轴突网络。利用二次谐波本征辐射（SHG）观察视网膜所有主要神经元的单轴突，即光感受器、水平细胞、双极细胞、无突细胞和视网膜神经节细胞。采用转基因GFP/YFP小鼠检测SHG+轴突细胞类型。视网膜SHG成像获得了新的发现：<s:1> ller细胞在过程中不保持均匀极化的微管；双极细胞的SHG+轴突以亚型特异性方式终止于内丛状层（IPL）；无毛细胞的一个子集，可能是轴突携带型，发出SHG；无毛细胞的轴突样神经突为IPL分层提供了细胞骨架支架。为了证明其实用性，我们以DBA/2小鼠为青光眼模型，DBA/2- gpnmb +小鼠为非青光眼对照组，应用视网膜SHG成像检测青光眼中视网膜内是否保留。结果发现青光眼视网膜内形态基本完整，视网膜神经节细胞的突触前区室未受损。这证明视网膜SHG成像技术是研究视网膜生理和病变的一种很有前途的3D技术。
We describe a novel method for visualizing the network of axons in the unlabeled fresh wholemount retina. The intrinsic radiation of second harmonic generation (SHG) was utilized to visualize single axons of all major retinal neurons, i.e., photoreceptors, horizontal cells, bipolar cells, amacrine cells, and the retinal ganglion cells. The cell types of SHG+ axons were determined using transgenic GFP/YFP mice. New findings were obtained with retinal SHG imaging: Müller cells do not maintain uniformly polarized microtubules in the processes; SHG+ axons of bipolar cells terminate in the inner plexiform layer (IPL) in a subtype-specific manner; a subset of amacrine cells, presumably the axon-bearing types, emits SHG; and the axon-like neurites of amacrine cells provide a cytoskeletal scaffolding for the IPL stratification. To demonstrate the utility, retinal SHG imaging was applied to testing whether the inner retina is preserved in glaucoma, using DBA/2 mice as a model of glaucoma and DBA/2-Gpnmb+ as the nonglaucomatous control. It was found that the morphology of the inner retina was largely intact in glaucoma and the presynaptic compartments to the retinal ganglion cells were uncompromised. It proves retinal SHG imaging as a promising technology for studying the physiological and diseased retinas in 3D.
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