Quantification of Catecholamine Uptake in Adult Cardiac Myocytes

In adult cardiac myocytes, multiple G protein-coupled receptors (GPCR) localize to and signal at the nucleus. These include endothelin B receptors, angiotensin type 1 and 2 receptors, β1- and β3-adrenergic receptors, and α1A- and α1B-adrenergic receptors. Initiation of signaling through nuclear GPCRs requires that ligands be produced within or transported into the cardiac myocytes, yet mechanisms whereby these ligands are produced or transported into cardiac myocytes are largely unclear. To activate nuclear adrenergic receptors in adult cardiac myocytes, uptake of endogenous catecholamines epinephrine and norepinephrine occurs via organic cation transporter 3 (OCT3), a member of the slc22a family of genes. This chapter details a method to detect and quantify catecholamine uptake in intact adult cardiac myocytes using a fl uorescent-based catecholamine uptake assay.