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  • Title: Intracellular mechanisms of cytoplasmic Ca2+ oscillation in rat megakaryocyte.
    Author: Uneyama H, Uneyama C, Akaike N.
    Journal: J Biol Chem; 1993 Jan 05; 268(1):168-74. PubMed ID: 7677993.
    Abstract:
    Extracellular application of ATP and ADP evoked the oscillatory K+ currents (IKCa) resulting from the periodic rise in cytoplasmic Ca2+ concentration ([Ca2+]i) of megakaryocyte isolated from rat bone marrow (Uneyama, H., Uneyama, C., and Akaike, N. (1992) Jpn. J. Pharmacol. 58, 231). The intracellular mechanism of ATP-induced cytoplasmic Ca2+ oscillation was investigated by the use of nystatin-perforated patch-clamp technique. Caffeine and ryanodine, which release Ca2+ from the Ca(2+)-induced Ca2+ release pool (CICR), and procaine, a blocker of Ca2+ release from CICR, had no effect on the IKCa oscillation in megakaryocyte. Thapsigargin and A23187 activated IKCa irreversibly by mobilizing Ca2+, but under Ca(2+)-free conditions they activated IKCa transiently. Intracellular application of inositol 1,4,5-triphosphate (IP3) also induced IKCa oscillation. Phorbol myristate acetate (PMA), a strong activator of protein kinase C (PKC), inhibited the oscillation completely. The inhibitory action of PMA was reversed by an inhibitor of PKC, staurosporin. The oscillation of ATP-induced IKCa was disrupted by staurosporin or calmodulin (CaM) antagonists such as W-7 and trifluoperazine, resulting in a transient and successive plateau-like IKCa. These results suggest that Ca2+ oscillation in megakaryocyte is caused by the interaction of both the Ca2+ release from IP3-sensitive Ca2+ pool and the Ca2+ uptake stimulated by PKC and Ca2+/CaM complex. Furthermore, forskolin, an activator of adenylate cyclase, and isobutylmethylxanthin, an inhibitor of phosphodiesterase, inhibited the frequency, latency, and current amplitude of the oscillation in a concentration-dependent manner. These reagents inhibited IP3-induced oscillation as well as an ATP-induced oscillation. Thus, the ATP-induced [Ca2+]i oscillation of rat megakaryocyte is also modulated by cAMP.
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