The plasma membrane Ca2+ ATPase (PMCA) is responsible for maintaining basal intracellular Ca2+ concentration ([Ca2+](i)) and returning small increases in [Ca2+](i) back to resting levels. The carboxyl terminus of some PMCA splice variants bind Homer proteins; how binding affects PMCA function is unknown. Here, we examined the effects of altered expression of Homer proteins on PMCA-mediated Ca2+ clearance from rat hippocampal neurons in culture. The kinetics of PMCA-mediated recovery from the [Ca2+](i) increase evoked by a brief train of action potentials was determined in the soma of single neurons using indo-1-based photometry. Exogenous expression of Homer la, Homer 1 c or Homer 2a did not affect PMCA function. However, shRNA mediated knockdown of Homer 1 slowed PMCA mediated Ca2+ clearance by 28% relative to cells expressing non-silencing shRNA. The slowed recovery rate in cells expressing Homer 1 shRNA was reversed by expression of a short Homer 2 truncation mutant. These results indicate that constitutively expressed Homer proteins tonically stimulate PMCA function in hippocampal neurons. We propose a model in which binding of short or long Homer proteins to the carboxyl terminus of the PMCA stimulates Ca2+ clearance rate. PMCA-mediated Ca2+ clearance may be stimulated following incorporation of the pump into Homer organized signaling domains and following induction of the Homer la immediate early gene. (C) 2012 Elsevier Inc. All rights reserved.