title: Contractile activation and measurements of intracellular Ca2+ concentration in cane toad twitch fibres in the presence of 2,3-butanedione monoxime.
abstract: The effects of 2,3-butanedione monoxime (BDM, 0.5-20 mM) on Ca(2+)-activated force in skinned muscle fibres, and force and Ca2+ responses in aequorin-injected intact fibres from the iliofibularis muscle of the cane toad Bufo marinus were investigated. Peak twitch force responses progressively decreased to 3% of the control response with increasing [BDM] up to 20 mM. Peak twitch aequorin light responses decreased to 65% of the control response in 10 mM BDM, but increased again to control values in 20 mM BDM. The duration of the twitch aequorin light response increased by up to 60% above 5 mM BDM. Tetanic (170 Hz) force and aequorin light responses reversibly decreased in a dose-dependent fashion to about 50% of the control response in 10 mM BDM. Failure of tetanic (170 Hz) stimulation was observed in the presence of 20 mM BDM. Intracellular [Ca2+] could be modified by changing the frequency of tetanic stimulation in the presence of BDM, permitting a study of the dependence of isometric force on intracellular [Ca2+] at different concentrations of BDM. In 10 mM BDM, the rate of force development in intact fibres was slower by a factor of two at saturating [Ca2+], and was up to one order of magnitude slower at non-saturating [Ca2+], when compared with control responses. At a similar intracellular [Ca2+] steady-state isometric force was reduced to about 85 and 50% of the control responses in 2 and 10 mM BDM, respectively. The effect of BDM on maximum Ca2+-activated force in skinned fibres paralleled the decrease in tetanic (170 Hz) force observed in intact fibres. The rate of force development in skinned fibres decreased with an increase in [BDM] at constant [Ca2+], and the sensitivity of the contractile apparatus to Ca2+ was shifted to a higher [Ca2+] by BDM. The results suggest that BDM reduces contractility in cane toad iliofibularis muscle by direct inhibition of the contractile apparatus, and reduction of the release of activator Ca2+ from the sarcoplasmic reticulum. Furthermore, BDM may be a useful tool to help study the relationship between force and [Ca2+] in intact muscle fibres.
authors.affiliation: Department of Zoology, La Trobe University, Bundoora, Victoria, Australia.
authors.forename: D J
authors.initials: DJ
authors.lastname: Lyster
authors.forename: D G
authors.initials: DG
authors.lastname: Stephenson
mesh_terms: D000331:Aequorin; D000818:Animals; D002024:Bufo marinus; D002118:Calcium / Q000378:metabolism*; D003931:Diacetyl / Q000031:analogs & derivatives* / Q000494:pharmacology; D006652:Histological Techniques; D007425:Intracellular Membranes / Q000378:metabolism*; D008163:Luminescent Measurements; D009119:Muscle Contraction / Q000187:drug effects*; D018485:Muscle Fibers, Skeletal / Q000187:drug effects* / Q000502:physiology*; D009994:Osmolar Concentration
chemical_list: C004717:diacetylmonoxime; D000331:Aequorin; D003931:Diacetyl; D002118:Calcium