Jia Xiaoyu, Feng Yibo, Ma Wenju, Zhao Wei, Liu Yanan, Jing Guangyin, Tian Jing, Yang Tao, Zhang Ce
State Key Laboratory of Photon-Technology in Western China Energy, Institute of Photonics and Photon-Technology, Northwest University, Shaanxi, Xi'an, China.
School of Physics, Northwest University, Shaanxi, Xi'an, China.
Front Mol Neurosci. 2023 Apr 6;16:1114928. doi: 10.3389/fnmol.2023.1114928. eCollection 2023.
Zebrafish is a suitable animal model for molecular genetic tests and drug discovery due to its characteristics including optical transparency, genetic manipulability, genetic similarity to humans, and cost-effectiveness. Mobility of the zebrafish reflects pathological conditions leading to brain disorders, disrupted motor functions, and sensitivity to environmental challenges. However, it remains technologically challenging to quantitively assess zebrafish's mobility in a flowing environment and simultaneously monitor cellular behavior .
We herein developed a facile fluidic device using mechanical vibration to controllably generate various flow patterns in a droplet housing single zebrafish, which mimics its dynamically flowing habitats.
We observe that in the four recirculating flow patterns, there are two equilibrium stagnation positions for zebrafish constrained in the droplet, i.e., the "source" with the outward flow and the "sink" with the inward flow. Wild-type zebrafish, whose mobility remains intact, tend to swim against the flow and fight to stay at the source point. A slight deviation from streamline leads to an increased torque pushing the zebrafish further away, whereas zebrafish with motor neuron dysfunction caused by lipin-1 deficiency are forced to stay in the "sink," where both their head and tail align with the flow direction. Deviation angle from the source point can, therefore, be used to quantify the mobility of zebrafish under flowing environmental conditions. Moreover, in a droplet of comparable size, single zebrafish can be effectively restrained for high-resolution imaging.
Using the proposed methodology, zebrafish mobility reflecting pathological symptoms can be quantitively investigated and directly linked to cellular behavior .
斑马鱼因其具有光学透明性、基因可操作性、与人类的基因相似性以及成本效益等特点，是分子遗传测试和药物发现的合适动物模型。斑马鱼的运动能力反映了导致脑部疾病、运动功能紊乱以及对环境挑战敏感性的病理状况。然而，在流动环境中定量评估斑马鱼的运动能力并同时监测细胞行为在技术上仍然具有挑战性。
我们在此开发了一种简便的流体装置，利用机械振动在容纳单个斑马鱼的液滴中可控地产生各种流动模式，该装置模拟了斑马鱼动态流动的栖息地。
我们观察到，在四种循环流动模式中，被限制在液滴中的斑马鱼有两个平衡停滞位置，即有向外流动的“源”和有向内流动的“汇”。运动能力未受影响的野生型斑马鱼倾向于逆流游动并努力停留在源点。与流线稍有偏差会导致推动斑马鱼进一步远离的扭矩增加，而因脂滴蛋白-1缺乏导致运动神经元功能障碍的斑马鱼则被迫停留在“汇”处，其头部和尾部均与流动方向对齐。因此，与源点的偏差角度可用于量化流动环境条件下斑马鱼的运动能力。此外，在大小相当的液滴中，单个斑马鱼可被有效限制以进行高分辨率成像。
使用所提出的方法，可以定量研究反映病理症状的斑马鱼运动能力，并将其与细胞行为直接联系起来。
