Changes in m6A demethylase ALKBH5 expression in form-deprivation myopic guinea pigs and its significance
10.3760/cma.j.cn115989-20231028-00156
- VernacularTitle:m6A去甲基化酶ALKBH5在形觉剥夺性近视豚鼠中的表达变化及其意义
- Author:
Xiaoling XIANG
1
;
Xuan LIAO
;
Jiayu MENG
;
Guimei ZHOU
;
Hong ZHOU
;
Xianmei ZHOU
Author Information
1. 川北医学院附属医院眼科 川北医学院眼视光医学院，南充 637000
- Keywords:
Myopia;
Guinea pigs;
AlkB homolog 5, RNA demethylase;
m6A methylation;
Pathogenesis
- From:
Chinese Journal of Experimental Ophthalmology
2024;42(4):315-321
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the alteration of m6A demethylase AlkB homolog 5 (ALKBH5) expression and its impact on form-deprivation myopia (FDM) retina in guinea pigs.Methods:Thirty normal SPF grade 3-week-old tricolor guinea pigs were randomly divided into normal control group and experimental group, with 15 in each group.In the experimental group, the right eyes were covered as FDM group and the left eyes uncovered were set as self-control group.Ocular biometry was performed at one-week intervals from baseline to week 4 of the experiment.Spherical equivalent was detected by streak retinoscopy and axial length was measured by A-scan ultrasonography.Animals were sacrificed after 4 weeks of modeling.The distribution and expression of ALKBH5 protein in the guinea pig retina was detected by immunohistochemical and immunofluorescence staining.Expression of ALKBH5 mRNA and protein in guinea pig retina was detected by real-time fluorescence quantitative PCR and Western blot, respectively.The use of animals in ophthalmic and vision research followed the tenets of Animal Research in Vision and Ophthalmology, and the study was approved by the Ethics Committee of North Sichuan Medical College (No.2023087).Results:At weeks 2, 3, and 4 after myopia induction, diopters and axial lengths were significantly higher in the FDM group than in the normal control group and the self-control group (all at P<0.001). Immunohistochemistry and immunofluorescence assays showed that ALKBH5 protein was expressed in the retinal nerve fiber layer, rod/cone photoreceptor cells, and retinal pigment epithelium (RPE) layer, and was highly expressed in the retinal nerve fiber layer and RPE layer.The relative ALKBH5 immunofluorescence intensity in the normal control group, self-control group and FDM group was 1.000±0.204, 0.874±0.076 and 0.571±0.053, respectively, which was lower in the FDM group than in the normal control and self-control groups, showing statistically significant differences ( t=4.069, P=0.006; t=5.176, P=0.014). After 4 weeks of modeling, ALKBH5 mRNA and protein expressions were significantly lower in FDM group than in normal control and self-control groups (both at P<0.01). Conclusions:The expression of m6A demethylase ALKBH5 is decreased in the retina of FDM guinea pigs, suggesting that ALKBH5 and related m6A methylation modification may be involved in the development and progression of myopia.
