Assessment of Swim Endurance and Swim Behavior in Adult Zebrafish - PubMed (original) (raw)

. 2021 Nov 12:(177):10.3791/63240.

doi: 10.3791/63240.

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Assessment of Swim Endurance and Swim Behavior in Adult Zebrafish

Brooke Burris et al. J Vis Exp. 2021.

Abstract

Due to their renowned regenerative capacity, adult zebrafish are a premier vertebrate model to interrogate mechanisms of innate spinal cord regeneration. Following complete transection of their spinal cord, zebrafish extend glial and axonal bridges across severed tissue, regenerate neurons proximal to the lesion, and regain their swim capacities within 8 weeks of injury. Recovery of swim function is thus a central readout for functional spinal cord repair. Here, we describe a set of behavioral assays to quantify zebrafish motor capacity inside an enclosed swim tunnel. The goal of these methods is to provide quantifiable measurements of swim endurance and swim behavior in adult zebrafish. For swim endurance, zebrafish are subjected to a constantly increasing water current velocity until exhaustion, and time at exhaustion is reported. For swim behavior assessment, zebrafish are subjected to low current velocities and swim videos are captured with a dorsal view of the fish. Percent activity, burst frequency, and time spent against the water current provide quantifiable readouts of swim behavior. We quantified swim endurance and swim behavior in wild-type zebrafish before injury and after spinal cord transection. We found that zebrafish lose swim function after spinal cord transection and gradually regain that capacity between 2 and 6 weeks post-injury. The methods described in this study could be applied to neurobehavioral, musculoskeletal, skeletal muscle regeneration, and neural regeneration studies in adult zebrafish.

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Conflict of interest statement

Disclosures

The authors have no conflicts of interest.

Figures

Figure 1:

Figure 1:. Swim tunnel set up and customized lids.

(A) Representative images of the swim tunnel set up including zoomed top and side views of the swim tunnel chamber. (B) Images of the swim tunnel lids used for the various applications described in this protocol. A standard, fully enclosed swim tunnel lid is used for swim behavior assays (section 3 of this protocol). A modified swim tunnel lid that accommodates a handheld digital flow meter is used for calibration (section 1 of this protocol). A modified swim endurance lid, containing a removable lid at the posterior end of the swim tunnel chamber, allows for the removal of exhausted fish during swim endurance testing (section 2 of this protocol).

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Figure 2:

Figure 2:. Experimental pipeline to assay for swim endurance and swim behavior in adult zebrafish.

For swim endurance, fish swam against an increasing water current until exhaustion. For swim behavior, swim parameters are assessed in the absence of and at low current velocities.

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Figure 3:

Figure 3:. Functional recovery in wild-type zebrafish after spinal cord injury.

(A) Motor function determined by swim endurance assays for wild-type zebrafish at baseline and 2, 4, and 6 wpi. Dots denote individual animals from two independent experiments. (B) Swim behavior assays tracked wild-type zebrafish under low water current velocities. The average Y position is shown at each time point throughout tracking (0 cm/s for 5 min, 10 cm/s for 5 min, and 20 cm/s for 5 min). (C–E) Percent activity (C), average Y position in the tunnel (D), and time swam against the flow (E) were quantified at 20 cm/s. For all quantifications, two independent experiments are shown. n = 30 in the pre-injury condition; n = 23 at 2 wpi, n = 20 at 4 wpi, n = 18 at 6 wpi. One-way ANOVA was used for statistical analyses. Error bars represent the Standard Error of the Mean (SEM). *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.

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