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Inflammation Research
Erschienen in: Inflammation Research 7/2011

01.07.2011 | Original Research Paper

Comparative analysis of inactivated-state block of N-type (Cav2.2) calcium channels

verfasst von: Timothy A. Vortherms, Andrew M. Swensen, Wende Niforatos, James T. Limberis, Torben R. Neelands, Richard S. Janis, Rama Thimmapaya, Diana L. Donnelly-Roberts, Marian T. Namovic, Di Zhang, C. Brent Putman, Ruth L. Martin, Carol S. Surowy, Michael F. Jarvis, Victoria E. Scott

Erschienen in: Inflammation Research | Ausgabe 7/2011

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Abstract

Objective

The aim of this study was to compare a diverse set of peptide and small-molecule calcium channel blockers for inactivated-state block of native and recombinant N-type calcium channels using fluorescence-based and automated patch-clamp electrophysiology assays.

Methods

The pharmacology of calcium channel blockers was determined at N-type channels in IMR-32 cells and in HEK cells overexpressing the inward rectifying K+ channel Kir2.1. N-type channels were opened by increasing extracellular KCl. In the Kir2.1/N-type cell line the membrane potential could be modulated by adjusting the extracellular KCl, allowing determination of resting and inactivated-state block of N-type calcium channels. The potency and degree of state-dependent inhibition of these blockers were also determined by automated patch-clamp electrophysiology.

Results

N-type-mediated calcium influx in IMR-32 cells was determined for a panel of blockers with IC50 values of 0.001–7 μM and this positively correlated with inactivated-state block of recombinant channels measured using electrophysiology. The potency of several compounds was markedly weaker in the state-dependent fluorescence-based assay compared to the electrophysiology assay, although the degree of state-dependent blockade was comparable.

Conclusions

The present data demonstrate that fluorescence-based assays are suitable for assessing the ability of blockers to selectively interact with the inactivated state of the N-type channel.
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Metadaten
Titel
Comparative analysis of inactivated-state block of N-type (Cav2.2) calcium channels
verfasst von
Timothy A. Vortherms
Andrew M. Swensen
Wende Niforatos
James T. Limberis
Torben R. Neelands
Richard S. Janis
Rama Thimmapaya
Diana L. Donnelly-Roberts
Marian T. Namovic
Di Zhang
C. Brent Putman
Ruth L. Martin
Carol S. Surowy
Michael F. Jarvis
Victoria E. Scott
Publikationsdatum
01.07.2011
Verlag
SP Birkhäuser Verlag Basel
Erschienen in
Inflammation Research / Ausgabe 7/2011
Print ISSN: 1023-3830
Elektronische ISSN: 1420-908X
DOI
https://doi.org/10.1007/s00011-011-0322-9

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