Department of Ophthalmology, Erasmus Medical Center, Rotterdam, the Netherlands.
Department of Clinical Genetics, Erasmus Medical Center, Rotterdam, the Netherlands.
Transl Vis Sci Technol. 2022 Mar 2;11(3):17. doi: 10.1167/tvst.11.3.17.
To establish a set of assays that allow the in vivo screening of candidate genes for ocular diseases in zebrafish, with an emphasis on refractive error.
Our pipeline includes the most relevant ocular screening measurements to assess (1) ocular biometry using spectral domain optical coherence tomography, (2) refractive status using an eccentric photorefractor, (3) intraocular pressure by tonometry, and (4) optokinetic response to study visual capability in zebrafish. To validate our pipeline and to demonstrate the potential of zebrafish as a valid animal model, we chose two well-characterized genes with an ocular phenotype (PRSS56 and FBN1) and generated two mutant zebrafish lines (prss56 and fbn1). Mutant fish were assessed at 2, 4, and 6 months after fertilization.
With the proposed phenotyping pipeline, we showed that ocular biometry, refractive status, intraocular pressure, and visual function can be studied in zebrafish. In the prss56 mutant, the pipeline revealed a dramatic decrease in axial length, mainly owing to a decreased vitreous chamber depth, whereas in the fbn1 mutant, ectopia lentis was the most distinctive ocular phenotype observed. Tonometry in both mutant lines showed an increase in intraocular pressure.
The proposed pipeline was applied successfully in zebrafish and can be used for future genetic screenings of candidate genes. While validating our pipeline, we found a close resemblance between the ocular manifestations in the zebrafish mutants and patients harboring mutations in PRSS56 and FBN1. Our results support the validity of our pipeline and highlight the potential of zebrafish as an animal model for in vivo screening of candidate genes for ocular diseases.
建立一套可用于在活体斑马鱼中筛选眼部疾病候选基因的检测方法，重点关注近视。
我们的方法包括了一系列最相关的眼部筛选测量方法，用于评估（1）使用谱域光相干断层扫描技术进行眼生物测量，（2）使用偏心光反射仪进行屈光状态评估，（3）通过眼压计进行眼内压测量，以及（4）使用视动反应评估斑马鱼的视觉能力。为了验证我们的方法并展示斑马鱼作为一种有效动物模型的潜力，我们选择了两个具有眼部表型的已知基因（PRSS56 和 FBN1），并生成了两个突变的斑马鱼系（prss56 和 fbn1）。在受精后 2、4 和 6 个月对突变鱼进行评估。
通过所提出的表型分析方法，我们表明可以在斑马鱼中研究眼生物测量、屈光状态、眼内压和视觉功能。在 prss56 突变体中，该方法揭示了轴向长度的显著减少，主要是由于玻璃体腔深度的减少，而在 fbn1 突变体中，晶状体异位是观察到的最显著的眼部表型。两种突变系的眼压测量均显示眼内压升高。
所提出的方法在斑马鱼中成功应用，并可用于未来候选基因的遗传筛选。在验证我们的方法的过程中，我们发现斑马鱼突变体的眼部表现与携带 PRSS56 和 FBN1 基因突变的患者非常相似。我们的结果支持我们的方法的有效性，并强调了斑马鱼作为一种用于活体筛选眼部疾病候选基因的动物模型的潜力。
