Cyclic GMP-activated channels of the chick pineal gland: effects of divalent cations, pH, and cyclic AMP.
Chick pineal cells maintained in dissociated cell culture express an intrinsic photosensitive circadian oscillator, but the mechanisms of phototransduction in avian pinealocytes are not fully understood. In this study, we have used inside-out patches to examine the characteristics of cyclic GMP-activated channels of chick pinealocytes in more detail, concentrating on the effects of factors known to modulate the secretion of melatonin and/or the function of circadian pacemakers. In most patches, the predominant conductance state was 19 pS in symmetrical 145 mM NaCl. But in some patches, a second cyclic GMP-activated channel with a unitary conductance of 29 pS was also present. The current flowing through cyclic GMP-activated channels was not affected by application of salines containing 1 microM Ca2+ to the cytoplasmic face of the patch membrane. By contrast, application of 1 mM Ca2+ caused a partial reduction in cyclic GMP-activated current at all membrane potentials. Application of 1-5 mM Mg2+ ions caused a virtually complete blockade of current at positive membrane potentials, but caused only a small decrease in current at negative membrane potentials. No obvious differences in the gating of cyclic GMP-activated channels were observed in pH 8.2, 7.4 or 6.2 salines. Application of salines containing 100 microM, 500 microM, or 1 mM cyclic AMP did not cause activation of the channels, but 5 mM cyclic AMP evoked a low level of channel activity. Application of 5 mM but not 100 microM cyclic AMP decreased the probability of channel activation caused by 20-100 microM cyclic GMP and also increased the percentage of openings to an 11 pS subconductance state. Thus, cyclic AMP acts as a weak partial agonist. Nevertheless, the gating of these channels does not seem to be controlled directly by physiologically relevant changes in intracellular Ca2+, pH, or cyclic AMP.
Dryer
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Department of Biological Science, Florida State University, Tallahassee 32306.
Henderson
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D000818:Animals; D002118:Calcium / Q000378:metabolism; D002413:Cations, Divalent / Q000494:pharmacology*; D002642:Chick Embryo; D002645:Chickens; D000242:Cyclic AMP / Q000378:metabolism / Q000494:pharmacology*; D006152:Cyclic GMP / Q000502:physiology*; D003593:Cytoplasm / Q000187:drug effects / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D015640:Ion Channel Gating / Q000187:drug effects / Q000502:physiology; D007473:Ion Channels / Q000502:physiology*; D008550:Melatonin / Q000378:metabolism; D008564:Membrane Potentials / Q000187:drug effects; D010870:Pineal Gland / Q000502:physiology*; D015398:Signal Transduction / Q000502:physiology
D002413:Cations, Divalent; D007473:Ion Channels; D000242:Cyclic AMP; D006152:Cyclic GMP; D008550:Melatonin; D002118:Calcium