National Clinical Research Center for Ocular Diseases, Eye Hospital, Wenzhou Medical University, Wenzhou 325027, China.
National Engineering Research Center of Ophthalmology and Optometry, Eye Hospital, Wenzhou Medical University, Wenzhou 325027, China.
Int J Mol Sci. 2024 Sep 5;25(17):9613. doi: 10.3390/ijms25179613.
Inherited retinal degenerations (IRDs) are a group of genetic disorders characterized by the progressive degeneration of retinal cells, leading to irreversible vision loss. has emerged as a candidate gene associated with IRDs, yet its mechanisms remain largely unknown. This study aims to investigate the role of in retinal development and its associated molecular pathogenesis in zebrafish. Morpholino oligonucleotide knockdown, CRISPR/Cas9 genome editing, quantitative RT-PCR, eye morphometric measurements, immunofluorescent staining, TUNEL assays, visual motor responses, optokinetic responses, rescue experiments, and bulk RNA sequencing were used to assess the impact of deficiency on retinal development. Our results demonstrated that the knockdown of resulted in a dose-dependent reduction in eye axial length, ocular area, and eye-to-body-length ratio. The fluorescence observations showed a significant decrease in immunofluorescence signals from photoreceptors and in mCherry fluorescence from RPE in -silenced morphants. TUNEL staining uncovered the extensive apoptosis of retinal cells induced by slc4a7 knockdown. Visual behaviors were significantly impaired in the -deficient larvae. GO and KEGG pathway analyses reveal that differentially expressed genes are predominantly linked to aspects of vision, ion channels, and phototransduction. This study demonstrates that the loss of in larvae led to profound visual impairments, providing additional insights into the genetic mechanisms predisposing individuals to IRDs caused by deficiency.
遗传性视网膜变性（IRDs）是一组以视网膜细胞进行性变性为特征的遗传疾病，导致不可逆转的视力丧失。 已成为与 IRDs 相关的候选基因，但其机制在很大程度上仍不清楚。本研究旨在探讨 在斑马鱼视网膜发育中的作用及其相关的分子发病机制。使用形态发生素寡核苷酸敲低、CRISPR/Cas9 基因组编辑、定量 RT-PCR、眼部形态测量、免疫荧光染色、TUNEL 测定、视觉运动反应、光动反应、挽救实验和批量 RNA 测序来评估 缺乏对视网膜发育的影响。我们的研究结果表明， 的敲低导致眼轴长度、眼面积和眼体长度比呈剂量依赖性降低。荧光观察显示，在沉默型形态发生素中，来自光感受器的免疫荧光信号和来自 RPE 的 mCherry 荧光显著减少。TUNEL 染色揭示了 slc4a7 敲低诱导的视网膜细胞广泛凋亡。 - 缺陷幼虫的视觉行为明显受损。GO 和 KEGG 途径分析表明，差异表达的基因主要与视觉、离子通道和光转导有关。本研究表明，幼虫中 的缺失导致严重的视觉障碍，为 缺乏引起的 IRDs 的遗传机制提供了更多的见解。
