THYROID-HORMONE ENHANCES CA2+ PUMPING ACTIVITY OF THE CARDIAC SARCOPLASMIC-RETICULUM BY INCREASING CA2+ ATPASE AND DECREASING PHOSPHOLAMBAN EXPRESSION

Phospholamban is a putative suppressor of the Ca2+ ATPase of the cardiac sarcoplasmic reticulum. The level of mRNA encoding the Ca2+ ATPase has been shown to be increased, whereas the phospholamban mRNA level to be decreased in the ventricles obtained from hyperthyroid rabbits [Nagai R, Zarain-Herzberg A, Brandl CJ, Fujii J, Tada M, MacLennan DH, Alpert NR, Periasamy M. (1989) Proc Natl Acad Sci USA 86: 2966-2970]. The present study was designed to examine whether these effects of thyroid hormone on the expression of the Ca2+ ATPase and phospholamban are exerted directly on cardiac myocytes and whether the resultant incoordinate expression of these proteins alters Ca2+ pumping activity. We studied the levels of phospholamban and Ca2+ ATPase mRNA in primary isolated neonatal rat myocardial cells incubated with triiodothyronine (T-3) for 3-48 h and the Ca2+ uptake activity of the microsomes prepared from the cells. Northern blot analysis showed that T-3 decreased phospholamban mRNA levels to about a half of control in 24 h. On the other hand, Ca2+ ATPase mRNA gradually increased with time. EC(50) for phospholamban mRNA expression was 2.5 x 10(-10) M which was approximately 10 times higher than that for the Ca2+ ATPase. T-3 increased V-max of Ca2+ uptake with the significant reduction of K-0.5 for Ca2+ (0.40 +/- 0.02 mu M for control v 0.31 +/- 0.02 mu M for T-3-treated vesicles), indicating that thyroid hormone stimulates Ca2+ pumping activity not only by increasing the Ca2+ ATPase but also decreasing phospholamban. These results suggested that phospholamban regulates the Ca2+ ATPase in dual modes; in short time range, by decreasing the affinity of the Ca2+ ATPase for Ca2+ by phosphorylation of phospholamban with cAMP-dependent protein kinase, and in long time range, by changing the molecular ratio between the two proteins through the regulation of gene expression.