Activation of L-type Ca2+ currents in cardiac myocytes by photoreleased GTP.
L-type calcium currents (ICa) were recorded from isolated ventricular myocytes by using standard patch-clamp methods. In the absence of agonist, photorelease of GTP by flash photolysis of intracellularly applied caged-GTP rapidly increased the amplitude of ICa over a wide range of membrane potentials. Control experiments clearly demonstrated that this effect was not due to either the release of photolytic by-products or to the light flash itself. The timecourse for activation of ICa by photolysis of caged-GTP was markedly altered by intracellular application of either GDP beta S or GTP gamma S. Upon maximal stimulation of ICa by intracellular dialysis with cAMP, photoreleased GTP induced a small, rapid increase in ICa followed by a gradual inhibition. The presence of Rp-cAMPS intracellularly reduced both the magnitude of the response to photoreleased GTP and its time to peak. Similar effects were observed when protein kinase inhibitor dialysed the cell interior, suggesting that both cAMP-dependent and independent processes were involved in this effect. We conclude that rapid release of GTP within ventricular myocytes, in the absence of agonist, causes rapid activation of L-type Ca2+ current. Mechanisms underlying this effect include stimulation of adenylate cyclase, together with other, as yet uncharacterized, GTP-dependent pathways for increasing ICa in the heart.
University Laboratory of Physiology, Oxford, U.K.
R Z
RZ
Kozlowski
L J
LJ
Goodstadt
V W
VW
Twist
T
T
Powell
D000818:Animals; D015220:Calcium Channels / Q000187:drug effects* / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology* / Q000528:radiation effects; D006168:Guinea Pigs; D066298:In Vitro Techniques; D007700:Kinetics; D009206:Myocardium / Q000378:metabolism*; D010782:Photolysis; D013873:Thionucleotides / Q000528:radiation effects; D014466:Ultraviolet Rays
D015220:Calcium Channels; D013873:Thionucleotides; C061507:P(3)-1-(2-nitro)phenylethylguanosine 5'-O-triphosphate; D006160:Guanosine Triphosphate; D019204:GTP-Binding Proteins