Acute effects of pregabalin on CaV2.1 calcium channels currents and cellular distribution in HEK293t cells

CARINA WEISSMANN; MARIANO N DI GUILMI; FRANCISCO J. URBANO; OSVALDO D. UCHITEL

BRAIN RESEARCH BULLETIN

PERGAMON-ELSEVIER SCIENCE LTD

Lugar: Amsterdam; Año: 2013 vol. 90 p. 107 - 113

0361-9230

We established a cell model to study the acute effects of pregabalin (PGB), a drug widely used in epilepsy and neuropathic pain, on voltage gated CaV2.1 (P/Q-type) calcium channels function and distribution at the membrane level. HEK293t cells were transfected with plasmids coding for all subunits of the CaV2.1 channel. We used a 1 fused to an eGFP tag to follow its distribution in time and at different experimental conditions. The expressed channel was functional as shown by the presence of barium-mediated, calcium currents of transfected cells measured by ?whole-cell voltage-clamp? recordings, showing a maximum current peak in the I?V curve at +20 mV. The GFP fluorescent signal was confined to the periphery of the cells. Incubation with 500 M PGB, that binds 2 subunits, for 30 min induced changes in localization of the fluorescent subunits as measured by fluorescent time lapse microscopy. These changes correlated with a reversible reduction of barium currents through CaV2.1 calcium channels under the same conditions. However, no changes in the cellular distribution of the subunits were visualized for cells either expressing another membrane associated protein or after exposure of the CaV2.1 channels to isoleucine, another 2 ligand. Together these results show strong evidence for an acute effect of PGB on CaV2.1 calcium channels? currents and distribution and suggest that internalization of CaV2.1 channels might be a mechanism of PGB action.