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调控人眼角膜上皮细胞中线粒体转移的机制。

Mechanisms Regulating Mitochondrial Transfer in Human Corneal Epithelial Cells.

机构信息

Department of Anatomy and Cell Biology, GW School of Medicine and Health Sciences, Washington, District of Columbia, United States.

Department of Ophthalmology, GW School of Medicine and Health Sciences, Washington, District of Columbia, United States.

出版信息

Invest Ophthalmol Vis Sci. 2024 Nov 4;65(13):10. doi: 10.1167/iovs.65.13.10.

DOI:10.1167/iovs.65.13.10
PMID:39504055
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11549930/
Abstract

PURPOSE

The intraepithelial corneal nerves (ICNs) innervating the cornea are essential to corneal epithelial cell homeostasis. Rho-associated kinase (ROCK) inhibitors (RIs) have been reported to play roles in neuron survival after injury and in mitochondrial transfer between corneal epithelial cells. In this study, the mechanisms human corneal limbal epithelial (HCLE) cells use to control intercellular mitochondrial transfer are assessed.

METHODS

Mitotracker and AAV1 mitotag eGFPmCherry were used to allow us to study mitochondrial transfer between HCLE cells and neurons in co-cultures and in HCLE cultures. A mitochondrial transfer assay was developed using HCLE cells to quantify the impact of cell stress and inhibition of phagocytosis, gap junctions, and ROCK on mitochondrial transfer, cell adhesion, migration, matrix deposition, and mitochondrial content.

RESULTS

Bidirectional mitochondrial transfer occurs between HCLE cells and neurons. Mitochondrial transfer among HCLE cells is inhibited when gap junction function is reduced and enhanced by acid stress and by inhibition of either phagocytosis or ROCK. Media conditioned by RI-treated cells stimulates cell adhesion and mitochondrial transfer.

CONCLUSIONS

Maximal mitochondrial transfer takes place when gap junctions are functional, when ROCK and phagocytosis are inhibited, and when cells are stressed by low pH media. Treatments that reduce mitochondrial content increase HCLE cell mitochondrial transfer. ROCK inhibition in co-cultures causes the release and adhesion of mitochondria to substrates where they can be engulfed by migrating HCLE cells and growing axons and their growth cones.

摘要

目的

角膜上皮内神经（ICN）支配角膜，对角膜上皮细胞稳态至关重要。 Rho 相关激酶（ROCK）抑制剂（RIs）已被报道在损伤后神经元存活和角膜上皮细胞之间的线粒体转移中发挥作用。在这项研究中，评估了人角膜缘上皮（HCLE）细胞控制细胞间线粒体转移的机制。

方法

使用 Mitotracker 和 AAV1 mitotag eGFPmCherry 允许我们研究 HCLE 细胞和神经元之间在共培养物和 HCLE 培养物中的线粒体转移。开发了一种线粒体转移测定法，使用 HCLE 细胞来量化细胞应激和吞噬作用、间隙连接和 ROCK 对线粒体转移、细胞黏附、迁移、基质沉积和线粒体含量的影响。

结果

HCLE 细胞和神经元之间发生双向线粒体转移。当降低间隙连接功能并通过酸应激和抑制吞噬作用或 ROCK 增强时，HCLE 细胞之间的线粒体转移受到抑制。经 RI 处理的细胞条件培养基刺激细胞黏附和线粒体转移。

结论

当间隙连接功能正常、ROCK 和吞噬作用受到抑制、低 pH 介质使细胞受到应激时，最大程度地发生线粒体转移。减少线粒体含量的治疗方法增加了 HCLE 细胞的线粒体转移。共培养物中 ROCK 抑制导致线粒体的释放和黏附到基质上，然后可以被迁移的 HCLE 细胞和生长轴突及其生长锥吞噬。

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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5800/11549930/2a273560de3f/iovs-65-13-10-f010.jpg
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