Downregulation of RyR and NCX in the neonatal rat ventricular myocyte modulates cytosolic [Ca2+]

Abstract

Calcium (Ca2+) is necessary for cardiac muscle contraction. RyR, NCX, and SERCA are key regulatory protein channels for cytosolic Ca2+ in cardiac myocytes. Expression levels of these proteins are a function of development, with protein expression shifting toward the adult phenotype over time. We investigated how downregulation by siRNAs of RyR and NCX affected expression levels of complimentary proteins and the corresponding intracellular Ca2+ transients. We compared experimentally observed Ca2+ transients to those predicted by mathematical models. Experiments show RyR downregulation decreased SERCA and increased NCX protein levels. The associated Ca2+ transient had a decreased amplitude, increased time-to-peak, 50%, and 90% Ca2+ removal with respect to the control cell. NCX downregulation increased SERCA production without significant changes in RyR expression levels. The corresponding [Ca2+] transient had increased amplitude, no change in time-to-peak and 50% Ca2+ removal, and increased 90% Ca2+ removal with respect to the control cell. Computational models that accurately predict the observed experimental data suggest compensatory changes occurring in the expression levels as well as biochemical activity of the regulatory proteins.