视网膜血管变性小鼠视网膜血管变性的特征和血管相关克劳丁蛋白的表达

IF 2 4区 医学 Q2 OPHTHALMOLOGY
Ophthalmic Research Pub Date : 2024-01-01 Epub Date: 2024-06-10 DOI:10.1159/000539605
Aoxiang Wang, Jinxi Zhou, Yiwen Hong, Yamei Cui, Yishen Wang, Jianying Pan, Yue Wu, Yan Luo
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引用次数: 0

摘要

引言本研究旨在探讨视网膜血管变性小鼠(Pde6βrd1/rd1 rd1小鼠)视网膜血管变性的特征及血管相关Claudin(CLD)蛋白的表达:方法:收集野生型(WT)小鼠和 rd1 小鼠出生后第 3 天(P3)、P5、P8、P11、P13、P15、P18 和 P21 天的视网膜。免疫荧光染色用于评估视网膜血管丛、细胞增殖、CLD表达和视网膜神经节细胞(RGC)。视网膜平片和冰冻切片的等选蛋白 B4 荧光染色可确定视网膜浅层和深层血管的分布。血红素和伊红染色以及末端脱氧核苷酸转移酶介导的 dNTP 缺口标记分别用于研究 rd1 小鼠视网膜组织学变性和细胞凋亡。利用实时定量 PCR 和 Western 印迹检测视网膜中血管相关的 CLD-1、2、3 和 CLD-5、血管内皮生长因子 A(VEGFA)和血管内皮生长因子受体 2(VEGFR2)的表达:结果:与 WT 小鼠相比,rd1 小鼠的视网膜浅层血管丛(SVP)和深层血管丛(DVP)发育延迟,但已完成渐进发育。在 rd1 小鼠中,视网膜层的厚度逐渐减少,视网膜逐渐萎缩和退化。这种退化在发育后期更为严重。SVP和DVP血管密度的下降与视网膜全层和内层厚度的下降以及RGC数量的减少有关。DVP的退化和核外层的变薄在P15时出现了明显的减少。Rd1小鼠的CLD-1、CLD-2、CLD-3、CLD-5、VEGFA和VEGFR2的表达水平下降,自P15起持续低于WT小鼠:Rd1小鼠在发育后期表现出视网膜SVP和DVP的进行性血管变性,视网膜ONL和RGC变薄和萎缩,血管相关的CLD蛋白下调。因此,rd1 小鼠不仅是视网膜神经变性的有用模型,也是视网膜血管变性的有用模型。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Characteristics of Retinal Vascular Degeneration and the Expression of Vessel-Related Claudin Proteins in Retinal Degeneration Mouse.

Introduction: This study aimed to investigate the characteristics of retinal vascular degeneration and the expression of vessel-related claudin (CLD) proteins in retinal degeneration mouse (Pde6βrd1/rd1 rd1 mouse).

Methods: Retinas from wild-type (WT) mice and rd1 mice at postnatal day 3 (P3), P5, P8, P11, P13, P15, P18, and P21 were collected. Immunofluorescence staining was used to assess the retinal vascular plexus, cell proliferation, CLD expression, and retinal ganglion cells (RGCs). The distribution of retinal superficial and deep vessels was determined by isolectin B4 fluorescence staining of retinal flat mounts and frozen sections. Hematoxylin and eosin staining and terminal deoxynucleotidyl transferase-mediated dNTP nick-end labeling were used to investigate retinal histological degeneration and apoptosis in rd1 mice, respectively. Quantitative real-time PCR and Western blot were used to measure the expression of vessel-related CLD-1, -2, -3, and -5, vascular endothelial growth factor A (VEGFA), and vascular endothelial growth factor receptor 2 (VEGFR2) in the retinas.

Results: Compared to the WT mice, the rd1 mice displayed delayed but completed progressive development in the retinal superficial vascular plexuses (SVPs) and deep vascular plexuses (DVPs). In the rd1 mice, the thickness of retinal layers gradually decreased and the retinas underwent progressive atrophy and degeneration. The deterioration got worse at the late developmental stage. The declined vessel density of SVP and DVP correlated with the decreased thickness of the full and inner parts of the retina and the reduced number of RGCs. DVP degeneration and the thinning of the outer nuclear layer exhibited an obvious reduction at P15. The expression levels of CLD-1, CLD-2, CLD-3, CLD-5, VEGFA, and VEGFR2 decreased and were consistently lower in the rd1 mice than in WT mice since P15.

Conclusion: Rd1 mice exhibited progressive vascular degeneration of retinal SVP and DVP, the thinning and atrophy of retinal ONL and RGC, and the downregulation of vessel-related CLD proteins during the late developmental period. Thus, the rd1 mouse is a useful model of not only retinal neuro-degeneration but also retinal vascular degeneration.

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来源期刊
Ophthalmic Research
Ophthalmic Research 医学-眼科学
CiteScore
3.80
自引率
4.80%
发文量
75
审稿时长
6-12 weeks
期刊介绍: ''Ophthalmic Research'' features original papers and reviews reporting on translational and clinical studies. Authors from throughout the world cover research topics on every field in connection with physical, physiologic, pharmacological, biochemical and molecular biological aspects of ophthalmology. This journal also aims to provide a record of international clinical research for both researchers and clinicians in ophthalmology. Finally, the transfer of information from fundamental research to clinical research and clinical practice is particularly welcome.
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