School of Medical Sciences, Mechanisms of Disease and Translational Research, University of New South Wales, Sydney, New South Wales, 2052, Australia.
Biomedical Imaging Facility, Mark Wainwright Analytical Centre, University of New South Wales, Sydney, New South Wales, 2052, Australia.
Ocul Surf. 2021 Jul;21:257-270. doi: 10.1016/j.jtos.2021.03.007. Epub 2021 Mar 22.
How sensory neurons and epithelial cells interact with one another, and whether this association can be considered an indicator of health or disease is yet to be elucidated.
Herein, we used the cornea, Confetti mice, a novel image segmentation algorithm for intraepithelial corneal nerves which was compared to and validated against several other analytical platforms, and three mouse models to delineate this paradigm. For aging, eyes were collected from 2 to 52 week-old normal C57BL/6 mice (n ≥ 4/time-point). For wound-healing and limbal stem cell deficiency, 7 week-old mice received a limbal-sparing or limbal-to-limbal epithelial debridement to their right cornea, respectively. Eyes were collected 2-16 weeks post-injury (n=4/group/time-point), corneas procured, immunolabelled with βIII-tubulin, flat-mounted, imaged by scanning confocal microscopy and analyzed for nerve and epithelial-specific parameters.
Our data indicate that nerve features are dynamic during aging and their curvilinear arrangement align with corneal epithelial migratory tracks. Moderate corneal injury prompted axonal regeneration and recovery of nerve fiber features. Limbal stem cell deficient corneas displayed abnormal nerve morphology, and fibers no longer aligned with corneal epithelial migratory tracks. Mechanistically, we discovered that nerve pattern restoration relies on the number and distribution of stromal-epithelial nerve penetration sites.
Microstructural changes to innervation may explain corneal complications related to aging and/or disease and facilitate development of new assays for diagnosis and/or classification of ocular and systemic diseases.
感觉神经元和上皮细胞如何相互作用，以及这种关联是否可以被视为健康或疾病的指标，目前仍有待阐明。
在此，我们使用角膜、Confetti 小鼠、一种新的用于上皮内角膜神经的图像分割算法，与其他几种分析平台进行了比较和验证，并使用三种小鼠模型来描绘这一范例。对于衰老，我们从 2 周到 52 周龄的正常 C57BL/6 小鼠中收集眼睛（n≥4/时间点）。对于伤口愈合和角膜缘干细胞缺乏，7 周龄的小鼠接受角膜缘保留或角膜缘到角膜缘上皮清创术，分别用于右眼。在损伤后 2-16 周（n=4/组/时间点）收集眼睛，采集角膜，用 βIII-微管蛋白免疫标记，平展，通过扫描共聚焦显微镜成像，并分析神经和上皮特异性参数。
我们的数据表明，神经特征在衰老过程中是动态的，它们的曲线排列与角膜上皮迁移轨迹一致。中度角膜损伤促使轴突再生和神经纤维特征的恢复。角膜缘干细胞缺乏的角膜显示出异常的神经形态，纤维不再与角膜上皮迁移轨迹对齐。从机制上讲，我们发现神经模式的恢复依赖于基质-上皮神经穿透点的数量和分布。
神经支配的微观结构变化可能解释与衰老和/或疾病相关的角膜并发症，并有助于开发用于诊断和/或分类眼部和系统性疾病的新检测方法。
