ZnT-1-induced ERK activation modulates T-type calcium channels and protect cardiomyocytes from ischemia reperfusion injury

Arie Moran 1 Yoram Etzion 1 Ofer Beharir 1 Shiri Levy 1 Merav Mor 1 Eden Shusterman 1 Daniel Gitler 1 Shani Dror 1 Joy Kahn 1 Amos Katz 2
1Physiology, Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer-Sheva
2Cardiology Department, Barzilai Medical Center, Ashkelon

Myocardial ischemia\infarction and heart failure are among the leading causes of morbidity and mortality worldwide. Activation of ERK signaling has been shown to promote cardioprotection from Ischemia-Reperfusion (I/R) Injury. ZnT-1, was found to interact with Raf-1 kinase, leading to downstream activation of ERK. In addition, we previously demonstrated that ZnT-1 inhibits L type calcium channels (LTCC) through interaction with the beta-subunit of the voltage-gated calcium channels. Here, we explored further the roles of ZnT-1 in the heart. Specifically, we studied the ability of ZnT-1 to protect cardiomyocytes from I/R injury. In addition, we explored the effect of ZnT-1 on T-type calcium channels (TTCC), which may have important role in the development of hypertrophy and automaticity in the diseased myocardium. In contrast to its inhibition of the LTCC, ZnT-1 stimulated TTCC currents and increased the surface expression of CaV3.1 (458 ±86 % of control, p<0.005). Inactive Raf-1 abolished ZnT-1 augmentation of the TTCC currents. In a model of I/R injury in HL-1 cells ZnT-1 increased Phospho-ERK and markedly reduced lactate dehydrogenase (LDH) release and activated caspase (programmed cell death marker). Conversely, knockdown of endogenous ZnT-1 by shRNA inhibited ERK phosphorylation and markedly increased LDH release following I/R. The MEK inhibitor PD98059 completely abolished the protective effect of ZnT-1. A truncated form of ZnT-1 lacking the C-terminal domain failed to induce ERK activation and did not protect the cells from I/R injury. In contrast, the C-terminal domain was sufficient to induce ERK activation and I/R protection. CONCLUSION: ZnT-1 stimulates the activity of the TTCC in a process involving ERK activation and increased TTCC surface expression. In addition to this role as a regulator of calcium homeostasis, ZnT-1 confers protection from I/R injury through its ability to activate ERK signaling. Thus, ZnT-1 seems to have important roles as a regulator of cardiac function.









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