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pubmed:abstractText |
1. The kinetic properties of the caffeine-induced transient outward current (ICaff) of the bull-frog sympathetic neurone were investigated using the extremely rapid concentration-jump technique. By setting the holding potential at the equilibrium potential for Cl- (-50 mV), the involvement of the Ca(2+)-activated Cl- current was suppressed. Using a Na(+)-free (Tris) external solution, the involvement of the Na(+)-dependent sustained outward current was eliminated. The 'M' conductance was also occluded by pre-treatment with muscarine. Under these experimental conditions, ICaff consisted of a TEA-sensitive Ca(2+)-activated K+ current. 2. When the latent period from the application of caffeine until the onset of ICaff (termed the ICaff latency) was measured, 10 mM-caffeine gave a latency of 10.5 +/- 0.7 ms (n = 14, mean +/- S.E.M.) at 22 degrees C. The latency was independent of caffeine concentration between 3 and 30 mM. 3. The ICaff latency was temperature-dependent; it was shortened when the temperature was elevated. 4. Both the time to peak and half-decay time of ICaff were decreased with increasing caffeine concentration. In each cell, these parameters decreased by increasing the amplitude of ICaff. 5. At 22 degrees C, the time to peak and the half-decay time of ICaff elicited by 10 mM-caffeine showed a linear relationship, and this relationship was preserved on either elevating or lowering the temperature. On lowering the temperature (12 degrees C), the time to peak shortened whereas the half-decay time was prolonged. On elevating the temperature (32 degrees C), the time to peak was prolonged whereas the half-decay time was shortened. 6. When EGTA in the intracellular solution was replaced by equimolar BAPTA, the time to peak was prolonged while the half-decay time was shortened. 7. It is concluded that caffeine can activate ICaff, with a time course in the order of milliseconds, and that the kinetics of activation and inactivation of ICaff reflect the time-dependent change in the total amount of intracellular free Ca2+.
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