Structure and function of skeletal muscle in zebrafish early larvae.
Zebrafish muscles were examined at an early developmental stage (larvae 5-7 d). Using aluminum clips, preparations (approximately 1.5 mm length, 150 microm diameter) were mounted for force registration and small angle x-ray diffraction. Sarcomeres were oriented mainly in parallel with the preparation long axis. Electrical stimulation elicited fast and reproducible single twitch contractions. Length-force relations showed an optimal sarcomere length of 2.15 microm. X-ray diffraction revealed clear equatorial 1.1/1.0 reflections, showing that myofilaments are predominantly arranged along the preparation long axis. In contrast, reflections from older (2 mo) zebrafish showed two main filament orientations each at an approximately 25 degrees angle relative to the preparation long axis. Electrical stimulation of larvae muscles increased the 1.1/1.0 intensity ratio, reflecting mass transfer to thin filaments during contraction. The apparent lattice volume was 3.42 x 10(-3) microm(3), which is smaller than that of mammalian striated muscle and more similar to that of frog muscles. The relation between force and stimulation frequency showed fusion of responses at a comparatively high frequency (approximately 186 Hz), reflecting a fast muscle phenotype. Inhibition of fast myosin with N-benzyl-p-toluene sulphonamide (BTS) showed that the later phase of the tetanus was less affected than the initial peak. This suggests that, although the main contractile phenotype is fast, slow twitch fibers can contribute to sustained contraction. A fatigue stimulation protocol with repeated 220 ms/186 Hz tetani showed that tetanic force decreased to 50% at a train rate of 0.1 s(-1). In conclusion, zebrafish larvae muscles can be examined in vitro using mechanical and x-ray methods. The muscles and myofilaments are mainly orientated in parallel with the larvae long axis and exhibit a significant fast contractile component. Sustained contractions can also involve a small contribution from slower muscle types.
Dou
Ying
Y
Department of Physiology and Pharmacology, Karolinska Institutet, Stockholm, Sweden.
Andersson-Lendahl
Monika
M
Arner
Anders
A
D008841:Actin Cytoskeleton / Q000378:metabolism; D000818:Animals; D000831:Animals, Newborn; D018360:Crystallography, X-Ray; D004558:Electric Stimulation; D007537:Isometric Contraction / Q000187:drug effects / Q000502:physiology; D007814:Larva / Q000033:anatomy & histology* / Q000502:physiology*; D018656:Muscle Fibers, Fast-Twitch / Q000502:physiology; D018657:Muscle Fibers, Slow-Twitch / Q000502:physiology; D053580:Muscle Strength / Q000187:drug effects / Q000502:physiology; D009129:Muscle Tonus / Q000187:drug effects / Q000502:physiology; D018482:Muscle, Skeletal / Q000502:physiology* / Q000648:ultrastructure*; D012518:Sarcomeres / Q000502:physiology / Q000648:ultrastructure; D013449:Sulfonamides / Q000494:pharmacology; D014050:Toluene / Q000031:analogs & derivatives / Q000494:pharmacology; D015027:Zebrafish / Q000033:anatomy & histology* / Q000196:embryology / Q000502:physiology*
C444880:N-benzyl-p-toluene sulfonamide; D013449:Sulfonamides; D014050:Toluene