Inhibition of Inositol(1,4,5)Trisphosphate Generation by Endothelin-1 During Postischemic Reperfusion

AbstractCalcium plays an integral role to many cellular processes including contraction, energy metabolism, gene expression, and cell death. The inositol 1,4,5-trisphosphate receptor (IP3R) is a calcium channel expressed in cardiac tissue. There are three IP3R isoforms encoded by separate genes. In the heart, the IP3R-2 isoform is reported to being most predominant with regards to expression levels and functional significance. The functional roles of IP3R-1 and IP3R-3 in the heart are essentially unexplored despite measureable expression levels. Here we show that all three IP3Rs isoforms are expressed in both neonatal and adult rat ventricular cardiomyocytes and in human heart tissue. All three IP3R proteins were expressed throughout the cardiomyocyte sarcoplasmic reticulum. Using isoform specific siRNA, we found that expression of all three IP3R isoforms are required for hypertrophic signaling downstream of endothelin-1 stimulation. Mechanistically, IP3Rs specifically contribute to activation of the hypertrophic program by mediating the positive inotropic effects of endothelin-1 leading to downstream activation of nuclear factor of activated T-cells. Our findings highlight previously unidentified functions for IP3R isoforms in the heart with significant implications for hypertrophic signaling in animal models and human disease.SignificanceHypertrophy is an adaptive response to cardiac stress which can lead to arrhythmias and cardiac failure. The peptide hormone endothelin-1(ET-1) is a potent activator of the hypertrophic program in cardiomyocytes. IP3R calcium channels are activated downstream of ET-1 during hypertrophy. We now show that all three IP3R proteins are essential for hypertrophic signaling downstream of ET-1. Activation of IP3Rs did not lead to nuclear-specific calcium transients but instead led to altered contractility ultimately, leading to NFAT activation and activation of the hypertrophic program. These effects were independent of alterations in IP3R protein expression levels both in vitro and in the human failing heart. Our results identify a new paradigm in IP3R signaling in the heart with relevance to human disease.

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Thapsigargin, a new inotropic agent, antagonizes action of endothelin-1 in rat atrial cells

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.1992.263.6.h1689 ◽

1992 ◽

Vol 263 (6) ◽

pp. H1689-H1694 ◽

Cited By ~ 1

Author(s):

P. Vigne ◽

J. P. Breittmayer ◽

C. Frelin

Keyword(s):

Sarcoplasmic Reticulum ◽

Inositol Phosphates ◽

Half Maximum ◽

Endothelin 1 ◽

Atrial Cells ◽

Inositol 1,4,5 Trisphosphate ◽

Slow Rise ◽

New Inotropic Agent ◽

Negative Inotropic Response ◽

Rat Atrial Cells

In isolated newborn rat atrial cells, thapsigargin induced a slow rise in cytosolic free Ca2+ concentration ([Ca2+]i) (half-maximum effective concentration = 1 microM) that was independent of the presence of external Ca2+. A 5-min treatment of atrial cells with 5 mM caffeine reduced but did not abolish the action of thapsigargin on [Ca2+]i. A first treatment of atrial cells with 10 microM thapsigargin reduced the action of ionomycin on [Ca2+]i. It also antagonized in a noncompetitive manner the Ca(2+)-mobilizing action of 100 nM endothelin-1 (ET-1). The half-maximum concentration for the inhibition by thapsigargin of ET-1 action was 0.2 microM. Thapsigargin had no action on the basal or ET-1 (100 nM)-stimulated production of inositol phosphates. These results suggest that thapsigargin discharges an inositol 1,4,5-trisphosphate-sensitive and caffeine-insensitive intracellular Ca2+ pool distinct from the sarcoplasmic reticulum. In isolated rat left atria, paced at 1 Hz, thapsigargin (10 microM) produced a transient 48% increase in contractility. It did not alter the contractile responses to 1 microM isoproterenol or to 5 mM caffeine. It had no action on postrest potentiation. Thapsigargin (10 microM) almost completely suppressed the positive inotropic action of 100 nM ET-1. It had no action on the transient negative inotropic response to ET-1. These results suggest that most of the positive inotropic effect of ET-1 is linked to its capacity to mobilize an inositol 1,4,5-trisphosphate-sensitive intracellular Ca2+ pool distinct from the sarcoplasmic reticulum.

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