Altered Ca2+handling by ryanodine receptor and Na+-Ca2+exchange in the heart from ovariectomized rats: role of protein kinase A

Abstract

Our previous study has demonstrated that ovariectomy (Ovx) significantly increased the left ventricular developed pressure (LVDP) and the maximal rate of developed pressure over time (±dP/d tmax) in the isolated perfused rat heart and the effects were reversed by female sex hormone replacement. In the present investigation, we studied the effects of Ovx for 6 wk on Ca2+homeostasis that determines the contractile function. Particular emphasis was given to Ca2+handling by ryanodine receptor (RyR) and Na+-Ca2+exchange (NCX).45Ca2+fluxes via the RyR, NCX, and Ca2+-ATPase (SERCA) were compared with their expression in myocytes from Ovx rats with and without estrogen replacement. Furthermore, we correlated the handling of Ca2+by these Ca2+handling proteins with the overall Ca2+homeostasis by determining the Ca2+transients induced by electrical stimulation and caffeine, which reveals the dynamic changes of cytosolic Ca2+concentration ([Ca2+]i) in the heart. In addition, we determined the expression and contribution of protein kinase A (PKA) to the regulation of the aforementioned Ca2+handling proteins in Ovx rats. It was found that after Ovx there were 1) increased Ca2+fluxes via RyR and NCX, which were reversed not only by estrogen replacement, but more importantly by blockade of PKA; 2) an increased expression of PKA; and 3) no increase in expression of NCX and SERCA. We suggest that hyperactivities of RyR and NCX are a result of upregulation of PKA. The increased release of Ca2+through RyR and removal of Ca2+by NCX are believed to be responsible for the greater contractility and faster relaxation after Ovx.