title: The effect of skeletal myosin light chain kinase gene ablation on the fatigability of mouse fast muscle.
abstract: Contraction-induced activation of a skeletal muscle specific Ca(2+) and calmodulin dependent myosin light chain kinase (skMLCK) catalyzes phosphorylation of the myosin regulatory light chain (RLC), a reaction that potentiates twitch force. The purpose of this study was to test the effect of skMLCK gene ablation on the fatigability of mouse extensor digitorum longus (EDL) muscle (in vitro at 25°C). Muscles were isolated from wildtype (WT, n = 10-12) and skeletal MLCK knockout (skMLCK KO, n = 10-12) mice and fatigued using a protocol consisting of 5 min of repeated tetanic stimulation (150 Hz for 1000 ms every 5 s). Both twitch (P(t)) and tetanic (P(o)) force as well as unloaded shortening velocity (V(o)) were assessed before, during and after fatiguing stimulation. Fatiguing stimulation increased RLC phosphorylation in WT but not skMLCK KO muscles (16 ± 0.01-0.63 ± 0.02 and 0.07 ± 0.02-0.08 ± 0.02 mol phos mol RLC, respectively). Although P(t) was potentiated above baseline in both WT and KO muscles, this increase was greater in WT than in KO muscles (to 1.37 ± 0.05 vs. 1.14 ± 0.02 of unpotentiated values, respectively). The difference in P(t) persisted until P(o) had been diminished to ~60% of baseline and thereafter P(t) declined to similar levels in both WT and KO muscles (to ~35% of initial). Overall, the time-course and decline in P(o) for WT and KO was similar (reduced to 0.20 ± 0.01 and 0.20 ± 0.01 of baseline, respectively) (P < 0.05). Initial values for V(o) were similar between WT and KO muscles and, moreover, the fatigue related decline in Vo was similar for both muscle genotypes (P < 0.05). Thus, our results demonstrate that skMLCK--catalyzed RLC phosphorylation augments isometric twitch force during moderate, but not severe, levels of fatigue.
authors.affiliation: Center for Muscle Metabolism and Biophysics, Faculty of Applied Health Sciences, Brock University, 500 Glenridge Ave, St Catharines, ON L2S 3A1, Canada.
authors.forename: William
authors.initials: W
authors.lastname: Gittings
authors.forename: Jiang
authors.initials: J
authors.lastname: Huang
authors.forename: Ian C
authors.initials: IC
authors.lastname: Smith
authors.forename: Joe
authors.initials: J
authors.lastname: Quadrilatero
authors.forename: Rene
authors.initials: R
authors.lastname: Vandenboom
mesh_terms: D000818:Animals; D055162:Biocatalysis; D017353:Gene Deletion; D007537:Isometric Contraction / Q000235:genetics / Q000502:physiology; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D018763:Muscle Fatigue / Q000235:genetics* / Q000502:physiology; D018994:Myosin Light Chains / Q000378:metabolism; D009219:Myosin-Light-Chain Kinase / Q000172:deficiency* / Q000235:genetics* / Q000378:metabolism; D010766:Phosphorylation
chemical_list: D018994:Myosin Light Chains; D009219:Myosin-Light-Chain Kinase