Effects of Topiramate on Kainate- and Domoate-Activated [14C]Guanidinium Ion Flux Through GluR6 Channels in Transfected BHK Cells Using Cytostar-T Scintillating Microplates

Authors


Address correspondence and reprint requests to Dr. R. P. Shank at The R. W. Johnson Pharmaceutical Research Institute, Spring House, PA 19477, U.S.A.

Summary:

Purpose: This study was undertaken to test the hypothesis that topiramate (TPM) exerts a negative modulatory effect on some types of α-amino-3-hydroxy-5-methyl-isoxazole-4-proprionic acid (AMPA)/kainate receptors by binding to the site at which protein kinase A (PKA) phosphory-lates the receptor-channel complex.

Methods: The effect of TPM on kainate- or domoate-induced [14C]guanidinium ion flux through iGluR6 channels expressed in baby hamster kidney (BHK) cells was evaluated. Because the hypothesis predicts that TPM will bind only in the dephos-phorylated state, a variety of experimental conditions were used to either promote or impede the phosphorylation of the receptor-channel complex. These included the use of dibutyryl cyclic adenosine monophosphate (cAMP) and forskolin to activate PKA, H-9 and H-89 to inhibit PKA, and okadaic acid to inhibit protein phosphatases.

Results: Kainate (1 μM) induced a gradual accumulation of [14C]guanidinium into the cells that plateaued ∼30 min after initiation of the reaction, whereas domoate (0·1 μM) caused a rapid accumulation into the cells that peaked within 5 min; thereafter, the amount of [14C]guanidinium in the cells declined gradually. Topiramate, at 0·1 and 100 μM, did not significantly affect the [14C]guanidinium accumulation under any of the experimental conditions used.

Conclusions: The results of this study are not consistent with the hypothesis tested. However, the results must be interpreted cautiously because iGluR6 receptors expressed in the BHK cells and the functional state of proteins that regulate AMPA/receptors (e.g., PSD-95) may not be sufficiently similar to the receptors and functional state in neurons to serve as a true test of the hypothesis.

Ancillary