We have used whole-cell recording techniques in the mouse neuroblastoma X rat glioma hybrid NG108-15 cell line to test whether protein kinase CK2 (CK2; also known as casein kinase II) modulates the function of the serotonin 5-HT(3) receptor (5-HT(3)R) channel. The rapid application of 5-HT (50 microM) to NG108-15 cells elicits a 5-HT(3)R-mediated inward current response that rapidly reaches peak amplitude and then desensitizes in the continued presence of agonist. Internal dialysis with CK2 (20 micrograms/ml) via the patch pipette significantly increases the amplitude and decreases the rate of desensitization of the 5-HT(3)R-mediated responses. CK2 that had been heat-inactivated has no effect on either the amplitude or the kinetics of desensitization of the 5-HT(3)R responses. These data suggest that dialysis with protein kinase CK2 significantly enhanced current through the 5-HT(3)R channel, and that CK2 may be an important regulator of 5-HT(3)R channel function in the nervous system, possibly serving to facilitate the 5-HT-induced excitation of the cells.