Using Touch-evoked Response and Locomotion Assays to Assess Muscle Performance and Function in Zebrafish

Abstract

Zebrafish muscle development is highly conserved with mammalian systems making them an excellent model to study muscle function and disease. Many myopathies affecting skeletal muscle function can be quickly and easily assessed in zebrafish over the first few days of embryogenesis. By 24 hr post-fertilization (hpf), wildtype zebrafish spontaneously contract their tail muscles and by 48 hpf, zebrafish exhibit controlled swimming behaviors. Reduction in the frequency of, or other alterations in, these movements may indicate a skeletal muscle dysfunction. To analyze swimming behavior and assess muscle performance in early zebrafish development, we utilize both touch-evoked escape response and locomotion assays.Touch-evoked escape response assays can be used to assess muscle performance during short burst movements resulting from contraction of fast-twitch muscle fibers. In response to an external stimulus, which in this case is a tap on the head, wildtype zebrafish at 2 days postfertilization (dpf) typically exhibit a powerful burst swim, accompanied by sharp turns. Our method quantifies skeletal muscle function by measuring the maximum acceleration during a burst swimming motion, the acceleration being directly proportional to the force produced by muscle contraction.In contrast, locomotion assays during early zebrafish larval development are used to assess muscle performance during sustained periods of muscle activity. Using a tracking system to monitor swimming behavior, we obtain an automated calculation of the frequency of activity and distance in 6-day old zebrafish, reflective of their skeletal muscle function. Measurements of swimming performance are valuable for phenotypic assessment of disease models and high-throughput screening of mutations or chemical treatments affecting skeletal muscle function.

A limb-girdle muscular dystrophy 2I model of muscular dystrophy identifies corrective drug compounds for dystroglycanopathies

“…In comparison, the dystrophic sapje zebrafish moved significantly more than the fkrp -/fish, although their overall distance traveled and velocity were reduced compared with control sibling zebrafish ( Figure 2, B and C). The touch-evoke response assay is another highly reproducible assay to access the overall abilities of a zebrafish larva to move their dorsal muscle in response to a physical stimulus (27,28). Touch-evoke analyses of the fkrp -/zebrafish revealed an overall decrease in response and movement compared with sibling wild-type and heterozygous controls ( Figure 2D).…”

“…DanioVision zebrafish motion tracking. Five-dpf zebrafish larvae were tracked via the DanioVision (Noldus) motion tracking platform in a protocol slightly modified from a previously published protocol (28). Zebrafish were placed in 48-well plates in 300 ml of fish water and were given a 15-minute acclimation period in the DanioVision machine.…”

Systemic loss of Sarm1 protects Schwann cells from chemotoxicity by delaying axon degeneration

“…Furthermore, a simple assay for sensorimotor function that consists of eliciting the escape response after tactile stimuli showed that the displacement distance and the average acceleration were no different between wild type and Sarm1 mutants ( Supplementary Fig. 1f, g) 45 .…”

L-tyrosine supplementation is not therapeutic for skeletal muscle dysfunction in zebrafish and mouse models of dominant skeletal muscle α-actin nemaline myopathy

“…Heart rate measurements were performed in triplicate with 10 fish per experiment. Assay of swimming ability, as well as the subsequent data analyses performed on 6 dpf wild type zebrafish treated with increasing doses of L-tyrosine, Tg ACTA1 D286G - eGFP and their control WT siblings treated with 10 µM L-tyrosine were as per (Sztal et al, 2016). Total voluntary distance travelled in a 10-minute period in the dark was measured in mm using zebraboxes (Viewpoint Life Sciences).…”