Optic nerve damage model: A preliminary study to determine injury duration and degree of gliosis

IF 2.8 3区医学 Q1 OPHTHALMOLOGY Acta Ophthalmologica Pub Date : 2025-01-19 DOI:10.1111/aos.16959

Humeyra Nur Kaleli, Cem Kesim, Murat Hasanreisoğlu

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Abstract

Aims/Purpose: The aim of this study was to investigate the effects of varying durations of optic nerve compression on retinal function and morphology in an adult DA/HanR rat with optic nerve crush model.

Methods: In the study, adult DA/HanR rats (4-8 mo) were divided into 5 experimental groups. Lateral canthotomy and lateral bulbar conjunctiva incision were performed to reach the area. The optic nerve was held 2-3 mm posterior to the globe with the help of calibrated forceps and compressed for four different periods of time: 10 seconds, 15 seconds, 30 seconds and 2 minutes. During the injury, retinal fundus observation of rats was checked with a 90-diopter fundus lens. To evaluate changes in vision after optic nerve damage, in vivo electrophysiological tests (VEP, EEG and ERG) with daylight stimulator, a light intensity of 500 μW/cm2, was studied. Changes in optic nerve and retina morphology after injury were evaluated by hematoxylin-eosin (H&E), TUNEL assay, immunofluorescence staining of gliosis and retina cell markers.

Results: It was observed that the response to light was reduced in both the retina (ERG) and visual cortex (VEP-EEG) in the damaged eyes compared to the control group. Retina cell survival showed decrease in injured groups by TUNEL assay. In immunofluorescence staining, an increase in the severity of gliosis with activation of the microglia and Müller cells in the retina and optic nerve depending on the duration of damage was shown by the expressions of GFAP, Glutamine Synthetase and Vimentin. It was observed that the expression of the neural marker NeuN and the photoreceptor marker Rhodopsin decreased upon injury.

Conclusions: Our study provided preliminary results to further optic nerve regeneration studies to select optimum time points for improve axonal healing in retina and optic nerve injuries.

Funding Information: This work is supported by the Scientific and Technological Research Council of Turkey (TÜBİTAK) under Grant ARDEB-121S762

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视神经损伤模型：确定神经胶质瘤损伤持续时间和程度的初步研究

目的：研究视神经压迫时间对视神经压迫模型DA/HanR大鼠视网膜功能和形态的影响。方法：将成年DA/HanR大鼠（4 ~ 8月龄）分为5个实验组。行外侧眦切开术及外侧球结膜切口到达该区域。在校准钳的帮助下，将视神经置于距眼球后2-3 mm处，并按压4个不同的时间段：10秒、15秒、30秒和2分钟。损伤期间用90屈光度眼底晶状体检查大鼠视网膜眼底观察。为评价视神经损伤后视力的变化，采用500 μW/cm2的日光刺激器进行体内电生理测试（VEP、EEG和ERG）。采用苏木精-伊红（H&；E）染色、TUNEL染色、胶质细胞免疫荧光染色及视网膜细胞标记物观察损伤后视神经和视网膜形态学的变化。结果：与对照组相比，损伤眼视网膜（ERG）和视觉皮层（VEP-EEG）对光的反应均降低。TUNEL法观察损伤组视网膜细胞存活率下降。在免疫荧光染色中，GFAP、谷氨酰胺合成酶和Vimentin的表达表明，随着视网膜和视神经中的小胶质细胞和突触细胞的激活，胶质细胞增生的严重程度随着损伤的持续时间而增加。观察到神经标记物NeuN和光感受器标记物Rhodopsin的表达在损伤后降低。结论：我们的研究为进一步的视神经再生研究提供了初步的结果，以选择最佳的时间点来改善视网膜和视神经损伤的轴突愈合。资助信息：本工作由土耳其科学技术研究委员会（TÜBİTAK）资助，基金编号为ARDEB-121S762

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来源期刊

Acta Ophthalmologica 医学-眼科学

CiteScore

7.60

自引率

5.90%

发文量

433

审稿时长

6 months

期刊介绍： Acta Ophthalmologica is published on behalf of the Acta Ophthalmologica Scandinavica Foundation and is the official scientific publication of the following societies: The Danish Ophthalmological Society, The Finnish Ophthalmological Society, The Icelandic Ophthalmological Society, The Norwegian Ophthalmological Society and The Swedish Ophthalmological Society, and also the European Association for Vision and Eye Research (EVER). Acta Ophthalmologica publishes clinical and experimental original articles, reviews, editorials, educational photo essays (Diagnosis and Therapy in Ophthalmology), case reports and case series, letters to the editor and doctoral theses.