Protein kinase A modulates A-type potassium currents of larval zebrafish (Danio rerio) white muscle fibres.
OBJECTIVE
Potassium (K(+)) channels are involved in regulating cell excitability and action potential shape. To our knowledge, very little is known about the modulation of A-type K(+) currents in skeletal muscle fibres. Therefore, we sought to determine whether K(+) currents of zebrafish white skeletal muscle were modulated by protein kinase A (PKA).


METHODS
Pharmacology and whole-cell patch clamp were used to examine A-type K(+) currents and action potentials associated with zebrafish white skeletal muscle fibres.


RESULTS
Activation of PKA by a combination of forskolin + 3-isobutyl-1-methylxanthine (Fsk + IBMX) decreased the peak current density by approximately 60% and altered the inactivation kinetics of A-type K(+) currents. The specific PKA inhibitor H-89 partially blocked the Fsk + IBMX-induced reduction in peak current density, but had no effect on the change in decay kinetics. Fsk + IBMX treatment did not shift the activation curve, but it significantly reduced the slope factor of activation. Activation of PKA by Fsk + IBMX resulted in a negative shift in the V(50) of inactivation. H-89 prevented all Fsk + IBMX-induced changes in the steady-state properties of K(+) currents. Application of Fsk + IBMX increased action potential amplitude, but had no significant effect on action potential threshold, half width or recovery rate, when fibres were depolarized with single pulses, paired pulses or with high-frequency stimuli.


CONCLUSIONS
PKA modulates the A-type K(+) current in zebrafish skeletal muscle and affects action potential properties. Our results provide new insights into the role of A-type K(+) channels in muscle physiology.
Coutts
C A
CA
Department of Biological Sciences, University of Alberta, Edmonton, AB, Canada.
Balt
L N
LN
Ali
D W
DW
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D015761:4-Aminopyridine / Q000494:pharmacology; D000200:Action Potentials / Q000187:drug effects; D000818:Animals; D005576:Colforsin / Q000494:pharmacology; D017868:Cyclic AMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000494:pharmacology*; D004625:Embryo, Nonmammalian / Q000378:metabolism; D005260:Female; D007546:Isoquinolines / Q000494:pharmacology; D008297:Male; D018656:Muscle Fibers, Fast-Twitch / Q000187:drug effects / Q000378:metabolism*; D018408:Patch-Clamp Techniques; D026902:Potassium Channel Blockers / Q000494:pharmacology; D024642:Potassium Channels, Voltage-Gated / Q000187:drug effects / Q000378:metabolism*; D013268:Stimulation, Chemical; D013449:Sulfonamides / Q000494:pharmacology; D015027:Zebrafish / Q000378:metabolism
D007546:Isoquinolines; D026902:Potassium Channel Blockers; D024642:Potassium Channels, Voltage-Gated; D013449:Sulfonamides; D005576:Colforsin; D015761:4-Aminopyridine; D017868:Cyclic AMP-Dependent Protein Kinases; C063509:N-(2-(4-bromocinnamylamino)ethyl)-5-isoquinolinesulfonamide; D015056:1-Methyl-3-isobutylxanthine