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  • Title: The ATP-induced inward current in mouse lacrimal acinar cells is potentiated by isoprenaline and GTP.
    Author: Sasaki T, Gallacher DV.
    Journal: J Physiol; 1992 Feb; 447():103-18. PubMed ID: 1375629.
    Abstract:
    1. ATP activates calcium (Ca2+) influx in mouse lacrimal acinar cells in the absence of phosphoinositide hydrolysis. Extracellular ATP (1 mM) activates receptor-operated cation channels, promoting entry of Na+ and Ca2+ (inward current). This Ca2+ influx in turn activates K+ channels resulting in a delayed, outward, current component. The present study uses patch-clamp current recording techniques to investigate the role of beta-adrenoceptor mechanisms, intracellular cyclic AMP and GTP in the regulation of the ATP-induced inward currents. 2. The beta-adrenoceptor agonist, isoprenaline (1 microM), does not increase the resting membrane currents but markedly enhances the ATP-induced inward and outward currents. This effect of isoprenaline is blocked by the beta-adrenoceptor antagonist propranolol. 3. Internal application of cyclic AMP mimics the potentiating effect of isoprenaline. 100 microM-cyclic AMP increases the ATP-induced inward and outward currents to about 200% as compared to control responses. 4. Pre-treatment of the cells with the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX; 1 mM), also results in a marked potentiation of the ATP-induced inward currents to 170% as compared to control responses. 5. The ATP-induced inward current responses are not blocked by either the removal of extracellular Ca2+ or by chelation of intracellular Ca2+ (by inclusion of 10 mM-EGTA in the recording pipette). Both protocols did however block the potentiating effect of internal cyclic AMP on the ATP-induced inward current responses. 6. Intracellular ATP (10 mM) reduces the amplitude of the inward currents evoked by external ATP application by about 60% and the currents were no longer potentiated by internal cyclic AMP. 7. Intracellular GTP or GTP-gamma-S (100 microM in the pipette solution) potentiates the current responses to ATP, increasing both the amplitude and duration of the inward currents. 8. In excised inside-out patches, with ATP in the recording pipette (i.e. external ATP), the catalytic subunit of the cyclic AMP-dependent protein kinase activated the cation channels. The effect of the catalytic subunit was readily reversible and abolished by an inhibitor of the protein kinase. 9. External ATP activates Ca2+ influx in lacrimal acinar cells by a mechanism that is distinct from that activated by phosphoinositide-coupled receptors. The effect is mediated by direct activation of cation channels in the cell surface membrane which allow for significant entry of Ca2+.(ABSTRACT TRUNCATED AT 400 WORDS)
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