Dynamic Imaging of Retinal Microglia

视网膜中的免疫细胞表现出显著的动态运动性，这反映了它们的生理和功能。 我们已经研究了眼免疫细胞的解剖和行为，包括1。 视网膜中的小胶质细胞，以及2. 脉络膜中的巨噬细胞 我们的工作采用了离体延时共聚焦成像技术来可视化来自转基因CX 3CR 1 +/GFP小鼠的荧光标记的小胶质细胞，并在真实的时间内跟踪完整视网膜和脉络膜外植体中的动态小胶质细胞行为。 脉络膜中的免疫和血管改变与年龄相关性黄斑变性（AMD）有关。由于脉络膜免疫细胞的生理学和与脉络膜血管的相互作用还不完全清楚，我们研究了年轻和老年小鼠脉络膜的髓样和血管成分之间的关系。 白化病CX 3CR 1（GFP/+）转基因小鼠的脉络膜髓样细胞具有绿色荧光，用活体染料DiI管腔内灌注以标记脉络膜血管。使用实时共聚焦成像和免疫组织化学分析来检查驻留骨髓细胞的分布、形态、行为和血管协会。 树突状髓样细胞，包括大多数的居民免疫细胞群体在脉络膜中，广泛分布在整个脉络膜，并表现出密切的协会与脉络膜血管，其在血管树中的位置不同。值得注意的是，与脉络膜动脉和小动脉相关的髓样细胞表现为位于血管长轴两侧的细长细胞，而与脉络膜毛细血管相关的髓样细胞在巩膜而不是玻璃体脉络膜毛细血管表面上分布为一层星状细胞。虽然静止的位置，树突状髓样细胞表现出快速的过程动力学非常适合全面的免疫监视血管周围的空间。髓样细胞的密度也随着年龄的增长而增加，与脉络膜血管衰减相关。 常驻骨髓细胞表现出密切的，但动态的物理相互作用与脉络膜血管，表明在正常脉络膜的组成性免疫血管相互作用。这些相互作用可能会随着年龄的增长而逐渐改变，为理解AMD的年龄相关脉络膜功能障碍提供了基础。

Immune cells in the retina demonstrate significant dynamic motility that report on their physiology and function. We have examined the anatomy and behavior of ocular immune cells including 1. microglia in the retina, and 2. macrophages in the choroid. Our work employed ex vivo time-lapse confocal imaging techniques to visualize fluorescence-labeled microglia from transgenic CX3CR1+/GFP mice and follow dynamic microglia behavior in intact retinal and choroidal explants in real time. Immune and vascular alterations in the choroid are implicated in age-related macular degeneration (AMD). As choroidal immune cells are incompletely understood with regard to their physiology and interactions with choroidal vessels, we examined the associations between myeloid and vascular components of the choroid in young and aged mice. Albino CX3CR1(GFP/+) transgenic mice, whose choroidal myeloid cells possess green fluorescence, were perfused intraluminally with the vital dye DiI to label choroidal vessels. The distribution, morphology, behavior, and vascular associations of resident myeloid cells were examined using time-lapse live confocal imaging and immunohistochemical analysis. Dendritiform myeloid cells, comprising most of the resident immune cell population in the choroid, were widely distributed across the choroid and demonstrated close associations with choroidal vessels that varied with their position in the vascular tree. Notably, myeloid cells associated with choroidal arteries and arterioles appeared as elongated cells flanking the long axes of vessels, whereas those associated with the choriocapillaris were distributed as a layer of stellate cells on the scleral but not vitreal choriocapillaris surface. While stationary in position, dendritiform myeloid cells demonstrated the rapid process dynamism well suited to comprehensive immunosurveillance of the perivascular space. Myeloid cells also increased in density as a function of aging, correlating locally with greater choroidal vascular attenuation. Resident myeloid cells demonstrated close but dynamic physical interactions with choroidal vessels, indicative of constitutive immune-vascular interactions in the normal choroid. These interactions may alter progressively with aging, providing a basis for understanding age-related choroidal dysfunction underlying AMD.