Troponin I phosphorylation in heart homogenate from diabetic rat

Although cardiac myofibrillar ATPase activity has been shown to be depressed during the development of diabetic heart dysfunction, the mechanisms of this alteration are not fully understood. Since phosphorylation of troponin I (TnI) is known to decrease the myofibrillar ATPase activity, the present study was undertaken to examine the TnI phosphorylation capacity in the diabetic heart homogenate. For this purpose rats were made diabetic by injecting streptozotocin (65 mg/kg; i.v.) and the hearts were removed 8 wk later. Some 6 wk diabetic animals were injected with insulin (3 U/day) for 2 wk. TnI content in the heart homogenate was measured by immunoblot assay, the mRNA abundance for TnI gene was determined by Northern blot analysis and the in vitro phosphorylation level of TnI was estimated by the ratio of phosphorylated TnI and total (phosphorylated and unphosphorylated) TnI. No significant changes in TnI content and gene expression of TnI were observed in right and left ventricles from the diabetic rats. However, the phosphorylation of TnI was higher (approximately 40%) in the diabetic hearts; this change was reversible upon insulin treatment. These results regarding TnI phosphorylation measured under in vitro conditions suggest that increased phosphorylation of TnI may contribute toward the depression in cardiac myofibrillar ATPase activity in chronic diabetes.