Abstract
HL-1 cells are the adult cardiac cell lines available that continuously divide while maintaining an atrial phenotype. Here we examined the expression and localization of inositol 1,4,5-trisphosphate receptor (IP(3)R) subtypes, and investigated how pattern of IP(3)-induced subcellular local Ca(2+) signaling is encoded by multiple IP(3)R subtypes in HL-1 cells. The type 1 IP(3)R (IP(3)R1) was expressed in the perinucleus with a diffuse pattern and the type 2 IP(3)R (IP(3)R2) was expressed in the cytosol with a punctate distribution. Extracellular ATP (1 mM) elicited transient intracellular Ca(2+) releases accompanied by a Ca(2+) oscillation, which was eliminated by the blocker of IP(3)Rs, 2-APB, and attenuated by ryanodine. Direct introduction of IP(3) into the permeabilized cells induced Ca(2+) transients with Ca(2+) oscillations at [Symbol: see text] 20 muM of IP(3), which was removed by the inhibition of IP(3)Rs using 2-APB and heparin. IP(3)-induced local Ca(2+) transients contained two distinct time courses: a rapid oscillation and a monophasic Ca(2+) transient. The magnitude of Ca(2+) oscillation was significantly larger in the cytosol than in the nucleus, while the monophasic Ca(2+) transient was more pronounced in the nucleus. These results provide evidence for the molecular and functional expression of IP(3)R1 and IP(3)R2 in HL-1 cells, and suggest that such distinct local Ca(2+) signaling may be correlated with the punctate distribution of IP(3)R2s in the cytosol and the diffuse localization of IP(3)R1 in the peri-nucleus.
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