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Abstract:

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 α1 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 +20mV. The GFP fluorescent signal was confined to the periphery of the cells. Incubation with 500μM PGB, that binds α2δ subunits, for 30min 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 α2δ 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. © 2012 Elsevier Inc.

Registro:

Documento: Artículo
Título:Acute effects of pregabalin on the function and cellular distribution of CaV2.1 in HEK293t cells
Autor:Weissmann, C.; Di Guilmi, M.N.; Urbano, F.J.; Uchitel, O.D.
Filiación:Insituto de Fisiología Biologia Molecular y Neurociencias (IFIBYNE-CONICET), University of Buenos Aires, Argentina
Palabras clave:CaV2.1 calcium channels; Epilepsy; HEK cells; Neuropathic pain; Pregabalin; barium; isoleucine; pregabalin; unclassified drug; voltage gated calcium channel; voltage gated calcium channel CaV2.1; article; calcium current; cellular distribution; controlled study; drug mechanism; electrophysiology; genetic transfection; microscopy; priority journal; protein expression; voltage clamp; Analgesics; Biophysics; Calcium Channels, N-Type; Electric Stimulation; gamma-Aminobutyric Acid; Green Fluorescent Proteins; HEK293 Cells; Humans; Ion Channel Gating; Membrane Potentials; Microscopy, Confocal; Patch-Clamp Techniques; Transfection
Año:2013
Volumen:90
Número:1
Página de inicio:107
Página de fin:113
DOI: http://dx.doi.org/10.1016/j.brainresbull.2012.10.001
Título revista:Brain Research Bulletin
Título revista abreviado:Brain Res. Bull.
ISSN:03619230
CODEN:BRBUD
CAS:barium, 7440-39-3; isoleucine, 7004-09-3, 73-32-5; pregabalin, 148553-50-8; Analgesics; Calcium Channels, N-Type; Green Fluorescent Proteins, 147336-22-9; enhanced green fluorescent protein; gamma-Aminobutyric Acid, 56-12-2; pregabalin; voltage-dependent calcium channel (P-Q type)
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_03619230_v90_n1_p107_Weissmann

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Citas:

---------- APA ----------
Weissmann, C., Di Guilmi, M.N., Urbano, F.J. & Uchitel, O.D. (2013) . Acute effects of pregabalin on the function and cellular distribution of CaV2.1 in HEK293t cells. Brain Research Bulletin, 90(1), 107-113.
http://dx.doi.org/10.1016/j.brainresbull.2012.10.001
---------- CHICAGO ----------
Weissmann, C., Di Guilmi, M.N., Urbano, F.J., Uchitel, O.D. "Acute effects of pregabalin on the function and cellular distribution of CaV2.1 in HEK293t cells" . Brain Research Bulletin 90, no. 1 (2013) : 107-113.
http://dx.doi.org/10.1016/j.brainresbull.2012.10.001
---------- MLA ----------
Weissmann, C., Di Guilmi, M.N., Urbano, F.J., Uchitel, O.D. "Acute effects of pregabalin on the function and cellular distribution of CaV2.1 in HEK293t cells" . Brain Research Bulletin, vol. 90, no. 1, 2013, pp. 107-113.
http://dx.doi.org/10.1016/j.brainresbull.2012.10.001
---------- VANCOUVER ----------
Weissmann, C., Di Guilmi, M.N., Urbano, F.J., Uchitel, O.D. Acute effects of pregabalin on the function and cellular distribution of CaV2.1 in HEK293t cells. Brain Res. Bull. 2013;90(1):107-113.
http://dx.doi.org/10.1016/j.brainresbull.2012.10.001