Chronic β-adrenergic stimulation reverses depressed Ca handling in mice overexpressing inhibitor-2 of protein phosphatase 1

Abstract

Rationale: A higher expression/activity of type 1 serine/threonine protein phosphatase 1 (PP1) may contribute to dephosphorylation of cardiac regulatory proteins triggering the development of heart failure.

Objective: Here, we tested the putatively protective effects of PP1 inhibitor-2 (I₂) overexpression using a heart failure model induced by chronic β-adrenergic stimulation.

Methods and results: Transgenic (TG) and wild-type (WT) mice were subjected to isoprenaline (ISO) or isotonic NaCl solution supplied via osmotic minipumps for 7 days. I₂ overexpression was associated with a depressed PP1 activity. Basal contractility was unchanged in catheterized mice and isolated cardiomyocytes between TG^NaCl and WT^NaCl. TG^ISO mice exhibited more fibrosis and a higher expression of hypertrophy marker proteins as compared to WT^ISO. After acute administration of ISO, the contractile response was accompanied by a higher sensitivity in TG^ISO as compared to WT^ISO. In contrast to basal contractility, the peak amplitude of [Ca]_i and SR Ca load were reduced in TG^NaCl as compared to WT^NaCl. These effects were normalized to WT levels after chronic ISO stimulation. Cardiomyocyte relaxation and [Ca]_i decay kinetics were hastened in TG^ISO as compared to WT^ISO, which can be explained by a higher phospholamban phosphorylation at Ser¹⁶. Chronic catecholamine stimulation was followed by an enhanced expression of GSK3β, whereas the phosphorylation at Ser⁹ was lower in TG as compared to the corresponding WT group. This resulted in a higher I₂ phosphorylation that may reactivate PP1.

Conclusion: Our findings suggest that the basal desensitization of β-adrenergic signaling and the depressed Ca handling in TG by inhibition of PP1 is restored by a GSK3β-dependent phosphorylation of I₂.

Keywords: Ca handling; Heart failure; Inhibitor-2; Protein phosphatase 1; β-adrenergic stimulation.