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施莱姆氏管内皮细胞连接在巨囊泡形成中的作用：三维电子显微镜研究。

The Role of Schlemm's Canal Endothelium Cellular Connectivity in Giant Vacuole Formation: A 3D Electron Microscopy Study.

机构信息

Department of Ophthalmology, Boston University School of Medicine, Boston, Massachusetts, United States.

The Affiliated Eye Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, China.

出版信息

Invest Ophthalmol Vis Sci. 2019 Apr 1;60(5):1630-1643. doi: 10.1167/iovs.18-26011.

DOI:10.1167/iovs.18-26011
PMID:30995299
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6736380/
Abstract

PURPOSE

We investigated whether cellular connectivity between Schlemm's canal (SC) inner wall (IW) endothelium, and juxtacanalicular connective tissue (JCT), and between IW endothelial cells, plays a role in giant vacuole (GV) and pore formation by comparing perfusion- and immersion-fixed eyes.

METHODS

Normal human donor eyes (n = 4) were either immersion-fixed (0 mm Hg) or perfusion-fixed (15 mm Hg). Trabecular meshwork near SC was imaged using serial block-face scanning electron microscopy. A total of 12 IW cells from each group were 3D-reconstructed from ∼7040 electron micrographs and compared. In each cell, connections between IW cells and JCT cells/matrix were quantified; IW/IW connectivity was measured by cell border overlap length. GV volume, density, shape, and intracellular and paracellular pores were analyzed.

RESULTS

The mean number of IW/JCT cell-cell connections per cell significantly decreased (P < 0.01) while the summed GV volume per cell significantly increased (P < 0.01) in perfusion-fixed eyes compared to immersion-fixed eyes. Intracellular pores were observed in 14.6% of GVs in perfusion-fixed eyes and not observed in immersion-fixed eyes. The mean IW/IW overlap length per cell decreased (P < 0.01), and paracellular pores were found only in regions where IW/IW connectivity was minimal (overlap length = 0 μm) in perfusion-fixed eyes and not observed in immersion-fixed eyes.

CONCLUSIONS

Our data suggest that changes in IW/JCT connectivity may be an important factor in the formation of larger GVs, and decreased IW/IW connectivity may promote paracellular pore formation. Targeting the IW/JCT and IW/IW connectivity may therefore be a potential strategy to regulate outflow resistance and IOP. .

摘要

目的

通过比较灌注固定和浸泡固定的眼睛，我们研究了施莱姆管（SC）内皮层（IW）内皮细胞与小梁网间隙连接组织（JCT）之间以及 IW 内皮细胞之间的细胞连接是否在巨泡（GV）和孔形成中起作用。

方法

正常的人供体眼球（n = 4）分别进行浸泡固定（0 毫米汞柱）或灌注固定（15 毫米汞柱）。使用连续块面扫描电子显微镜对 SC 附近的小梁网进行成像。从每组中总共重建了 12 个 IW 细胞，从大约 7040 张电子显微镜照片中进行了比较。在每个细胞中，IW 细胞与 JCT 细胞/基质之间的连接数量进行了量化；通过细胞边界重叠长度测量 IW/IW 连接性。分析了 GV 体积、密度、形状以及细胞内和细胞旁孔。

结果

与浸泡固定的眼睛相比，灌注固定的眼睛中每个细胞的 IW/JCT 细胞-细胞连接数量显著减少（P < 0.01），而每个细胞的 GV 体积总和显著增加（P < 0.01）。在灌注固定的眼睛中观察到 14.6%的 GV 中存在细胞内孔，而在浸泡固定的眼睛中未观察到。每个细胞的 IW/IW 重叠长度平均值减少（P < 0.01），并且仅在灌注固定的眼睛中 IW/IW 连接性最小（重叠长度= 0 μm）的区域中发现细胞旁孔，而在浸泡固定的眼睛中未观察到。

结论

我们的数据表明，IW/JCT 连接性的变化可能是形成更大 GV 的一个重要因素，而 IW/IW 连接性的降低可能会促进细胞旁孔的形成。因此，靶向 IW/JCT 和 IW/IW 连接性可能是调节流出阻力和 IOP 的潜在策略。

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