Power Derek, Elstrott Justin, Schallek Jesse
Center for Visual Science, University of Rochester, Rochester, United States.
Flaum Eye Institute, University of Rochester, Rochester, United States.
Elife. 2025 Jul 22;13:RP98662. doi: 10.7554/eLife.98662.
In response to central nervous system (CNS) injury, tissue-resident immune cells such as microglia and circulating systemic neutrophils are often first responders. The degree to which these cells interact in response to CNS damage is poorly understood, and even less so, in the neural retina, which poses a challenge for high-resolution imaging in vivo. In this study, we deploy fluorescence adaptive optics scanning light ophthalmoscopy (AOSLO) to study microglia and neutrophils in mice. We simultaneously track immune cell dynamics using label-free phase-contrast AOSLO at micron-level resolution. Retinal lesions were induced with 488 nm light focused onto photoreceptor (PR) outer segments. These lesions focally ablated PRs, with minimal collateral damage to cells above and below the plane of focus. We used in vivo AOSLO, and optical coherence tomography (OCT) imaging to reveal the natural history of the microglial and neutrophil response from minutes to months after injury. While microglia showed dynamic and progressive immune response with cells migrating into the injury locus within 1 day after injury, neutrophils were not recruited despite close proximity to vessels carrying neutrophils only microns away. Post-mortem confocal microscopy confirmed in vivo findings. This work illustrates that microglial activation does not recruit neutrophils in response to acute, focal loss of PRs, a condition encountered in many retinal diseases.
针对中枢神经系统(CNS)损伤,诸如小胶质细胞等组织驻留免疫细胞和循环系统中的中性粒细胞通常是首批响应者。人们对这些细胞在中枢神经系统损伤时相互作用的程度了解甚少,而在神经视网膜中更是知之甚少,这给体内高分辨率成像带来了挑战。在本研究中,我们采用荧光自适应光学扫描激光检眼镜(AOSLO)来研究小鼠中的小胶质细胞和中性粒细胞。我们使用无标记相衬AOSLO以微米级分辨率同时追踪免疫细胞动态。用聚焦于光感受器(PR)外段的488nm光诱导视网膜损伤。这些损伤局部消融PR,对焦点平面上方和下方的细胞造成的附带损伤最小。我们使用体内AOSLO和光学相干断层扫描(OCT)成像来揭示损伤后数分钟至数月内小胶质细胞和中性粒细胞反应的自然病程。虽然小胶质细胞表现出动态且渐进的免疫反应,细胞在损伤后1天内迁移至损伤部位,但尽管与仅几微米远的携带中性粒细胞的血管距离很近,中性粒细胞并未被募集。死后共聚焦显微镜检查证实了体内研究结果。这项工作表明,在许多视网膜疾病中出现的PR急性局灶性丧失的情况下,小胶质细胞激活不会募集中性粒细胞。
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