Gómez Sánchez Alicia, Álvarez Yolanda, Colligris Basilio, Kennedy Breandán N
Ocupharm Diagnostic Group Research, Faculty of Optic and Optometry, Universidad Complutense de Madrid, Madrid, Spain.
UCD Conway Institute of Biomolecular and Biomedical Research, University College Dublin, Belfield, Dublin 4, Ireland.
Open Res Eur. 2022 Jan 6;1:92. doi: 10.12688/openreseurope.13923.2. eCollection 2021.
The optokinetic response (OKR) is an effective behavioural assay to investigate functional vision in zebrafish. The rapid and widespread use of gene editing, drug screening and environmental modulation technologies has resulted in a broader need for visual neuroscience researchers to access affordable and more sensitive OKR, contrast sensitivity (CS) and visual acuity (VA) assays. Here, we demonstrate how 2D- and 3D-printed, striped patterns or drums coupled with a motorised base and microscope provide a simple, cost-effective but efficient means to assay OKR, CS and VA in larval-juvenile zebrafish. In wild-type, five days post-fertilisation (dpf) zebrafish, the 2D or 3D set-ups of 0.02 cycles per degree (cpd) (standard OKR stimulus) and 100% black-white contrast evoked equivalent responses of 24.2±3.9 or 21.8±3.9 saccades per minute, respectively. Furthermore, although the OKR number was significantly reduced compared to the 0.02 cpd drum (p<0.0001), 0.06 and 0.2 cpd drums elicited equivalent responses with both set-ups. Notably, standard OKRs varied with time of day; peak responses of 29.8±7 saccades per minute occurred in the early afternoon with significantly reduced responses occurring in the early morning or late afternoon (18.5±3 and 18.4±4.5 saccades per minute, respectively). A customised series of 2D printed drums enabled analysis of VA and CS in 5-21 dpf zebrafish. The saccadic frequency in VA assays was inversely proportional to age and spatial frequency and in CS assays was inversely proportional to age and directly proportional to contrast of the stimulus. OKR, VA and CS of zebrafish larvae can be efficiently measured using 2D- or 3D-printed striped drums. For data consistency the luminance of the OKR light source, the time of day when the analysis is performed, and the order of presentation of VA and CS drums must be considered. These simple methods allow effective and more sensitive analysis of functional vision in zebrafish.
视动反应（OKR）是一种用于研究斑马鱼视觉功能的有效行为学检测方法。基因编辑、药物筛选和环境调控技术的迅速广泛应用，使得视觉神经科学研究人员对获取经济实惠且更灵敏的OKR、对比敏感度（CS）和视敏度（VA）检测方法的需求更为广泛。在此，我们展示了如何通过二维和三维打印的条纹图案或鼓，结合电动底座和显微镜，为检测幼体-幼年期斑马鱼的OKR、CS和VA提供一种简单、经济高效的方法。在野生型受精后5天（dpf）的斑马鱼中，每度0.02周/度（cpd）（标准OKR刺激）的二维或三维设置以及100%的黑白对比度分别引发了每分钟24.2±3.9次或21.8±3.9次扫视的等效反应。此外，尽管与0.02 cpd的鼓相比，OKR次数显著减少（p<0.0001），但0.06和0.2 cpd的鼓在两种设置下引发的反应相当。值得注意的是，标准OKR随一天中的时间而变化；每分钟29.8±7次扫视的峰值反应出现在下午早些时候，而在清晨或傍晚（分别为每分钟18.5±3次和18.4±4.5次扫视）反应显著减少。一系列定制的二维打印鼓能够分析5-21 dpf斑马鱼的VA和CS。VA检测中的扫视频率与年龄和空间频率成反比，而在CS检测中与年龄成反比且与刺激对比度成正比。使用二维或三维打印的条纹鼓可以有效地测量斑马鱼幼体的OKR、VA和CS。为了保证数据的一致性，必须考虑OKR光源的亮度、进行分析的时间以及VA和CS鼓的呈现顺序。这些简单的方法能够对斑马鱼的视觉功能进行有效且更灵敏的分析。
