Multi-phenotypic and bi-directional behavioral screening of zebrafish larvae

Abstract

Multi-phenotypic screening of zebrafish larvae, such as monitoring the heart and tail activities, is important in biological assays. Microfluidic devices have been developed for zebrafish phenotypic assays, but simultaneous lateral–dorsal screening of the same larva in a single chip is yet to be achieved. We present a multi-phenotypic microfluidic device for monitoring of tail movement and heart rate (HR) of 5–7-day postfertilization zebrafish larvae. Tail movements were stimulated using electric current and quantified in terms of response duration (RD) and tail beat frequency (TBF). The positioning of a right-angle prism provided a lateral view of the larvae and enabled HR monitoring. Investigations were performed on zebrafish larvae exposed to 3% ethanol, 250 μM 6-hydroxydopamine (6-OHDA) or 1 mM levodopa. Larvae exposed to ethanol showed a significant drop in HR, whereas electric stimulation increased the HR temporarily. Larvae experienced a significant drop in RD, TBF and HR when exposed to 6-OHDA. HR was not affected by levodopa post-treatment, whereas RD and TBF were restored to normal levels. The results showed potential for applications that involve monitoring of cardiac and behavioral parameters in zebrafish larvae. Tests can be done using the same chip, without changing the larvae’s orientation. This eliminates undue stress caused by reorientation, which may affect their behavior, and the use of separate devices to obtain dorsal and lateral views. The device can be implemented to improve multi-phenotypic and quantitative screening of zebrafish larvae in response to chemical and physical stimuli in different zebrafish disease models.

Loss of Panx1 function in zebrafish alters motor behavior in a lab‐on‐chip model of Parkinson's disease

“…The valve in the bottom layer was then pressurized, causing the middle membrane to deflect and create a physical barrier preventing the larva from swimming out of the TR. Following a 1-min recovery period, shown to be sufficient to return cardiac activity to baseline, the electric stimulus of 3 μA was applied for 20 s using the sourcemeter (Khalili et al, 2020). The larva's locomotor response could be recorded with a camera at 2× magnification on the Leica stereomicroscope.…”

“…We previously demonstrated visual and motor deficiencies in this fish line (Safarian et al, 2020). We also have reported different microfluidic techniques to study the electric‐induced behavioral responses of zebrafish larvae, quantitatively (Khalili et al, 2019, 2020; Khalili, van Wijngaarden, Youssef, et al, 2021; Khalili, van Wijngaarden, Zoidl, et al, 2021; Peimani et al, 2018). The lab‐on‐chip approach allows us to address the challenges of behavioral studies such as providing a controllable stimulus to evoke the behavioral responses of larvae and quantifying their quick movements.…”

Loss of Panx1 Function in Zebrafish Alters Motor Behavior in a Lab-on-Chip Model of Parkinson's Disease

“…The valve, situated in the bottom layer, was then pressurized, causing the middle membrane to deflect and create a physical barrier that prevented the larva from swimming out of the TR. Following a one-minute recovery period, shown to be sufficient to return cardiac activity to baseline, the electric stimulus of 3μA was applied for 20 s using the sourcemeter [39]. The larva's locomotor response could be recorded with a camera at 2x magnification on the Leica stereomicroscope (Fig.…”

“…Here, an association of Panx1 and PD was investigated by comparing molecular and behavioral properties of Panx1a knockout (KO) (panx1a -/-) and wild type (WT) (panx1a +/+ ) zebrafish using the 6-OHDA model. We have previously reported different microfluidic techniques to study the electric-induced behavioral responses of zebrafish larvae, quantitatively [20], [36]- [39]. The lab-on-chip approach allows to address challenges of behavioral studies such as providing a controllable stimulus to evoke the behavioral responses of larvae and quantifying their quick movements.…”

Zebrafish Larvae's Response to Electricity is Mediated by Dopaminergic Agonists and Antagonists

“…Zebrafish larvae offer several advantages for behavioural studies applicable in examining and creating treatments to combat neurodegenerative and psychiatric disorders. Many microfluidic devices have been designed for manipulation of zebrafish embryos and larvae [1][2][3][4][5][6][7][8] and stimulating them [9][10][11][12][13][14][15][16] to gather information about their neural [15,17] and behavioral [13][14][15][16]18,19] activities in controlled microenvironments. Various stimuli such as thermal [9] , chemical [20] and fluid flow [11] have been tested.…”