caa a22 4500 44588522X CHVBK 20180317145557.0 cr unu---uuuuu 170323e20110701xx s 000 0 eng 10.1007/s00011-011-0322-9 doi (NATIONALLICENCE)springer-10.1007/s00011-011-0322-9 Comparative analysis of inactivated-state block of N-type (Cav2.2) calcium channels [Elektronische Daten] [Timothy Vortherms, Andrew Swensen, Wende Niforatos, James Limberis, Torben Neelands, Richard Janis, Rama Thimmapaya, Diana Donnelly-Roberts, Marian Namovic, Di Zhang, C. Brent Putman, Ruth Martin, Carol Surowy, Michael Jarvis, Victoria Scott] 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 0.001-7μM 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. Springer Basel AG, 2011 N-type calcium channels nationallicence Cav2.2 nationallicence State-dependent inhibition nationallicence Fluorometric imaging plate reader (FLIPR) nationallicence Automated patch clamp electrophysiology nationallicence QPatch HT nationallicence Vortherms Timothy Neuroscience Research, Global Pharmaceutical Research and Development, Abbott Laboratories, Dept R4PM, Bldg. AP9A, 100 Abbott Park Road, 60064-6125, Abbott Park, IL, USA aut Swensen Andrew Neuroscience Research, Global Pharmaceutical Research and Development, Abbott Laboratories, Dept R4PM, Bldg. AP9A, 100 Abbott Park Road, 60064-6125, Abbott Park, IL, USA aut Niforatos Wende Neuroscience Research, Global Pharmaceutical Research and Development, Abbott Laboratories, Dept R4PM, Bldg. AP9A, 100 Abbott Park Road, 60064-6125, Abbott Park, IL, USA aut Limberis James Neuroscience Research, Global Pharmaceutical Research and Development, Abbott Laboratories, Dept R4PM, Bldg. AP9A, 100 Abbott Park Road, 60064-6125, Abbott Park, IL, USA aut Neelands Torben Neuroscience Research, Global Pharmaceutical Research and Development, Abbott Laboratories, Dept R4PM, Bldg. AP9A, 100 Abbott Park Road, 60064-6125, Abbott Park, IL, USA aut Janis Richard Neuroscience Research, Global Pharmaceutical Research and Development, Abbott Laboratories, Dept R4PM, Bldg. AP9A, 100 Abbott Park Road, 60064-6125, Abbott Park, IL, USA aut Thimmapaya Rama Neuroscience Research, Global Pharmaceutical Research and Development, Abbott Laboratories, Dept R4PM, Bldg. AP9A, 100 Abbott Park Road, 60064-6125, Abbott Park, IL, USA aut Donnelly-Roberts Diana Neuroscience Research, Global Pharmaceutical Research and Development, Abbott Laboratories, Dept R4PM, Bldg. AP9A, 100 Abbott Park Road, 60064-6125, Abbott Park, IL, USA aut Namovic Marian Neuroscience Research, Global Pharmaceutical Research and Development, Abbott Laboratories, Dept R4PM, Bldg. AP9A, 100 Abbott Park Road, 60064-6125, Abbott Park, IL, USA aut Zhang Di Neuroscience Research, Global Pharmaceutical Research and Development, Abbott Laboratories, Dept R4PM, Bldg. AP9A, 100 Abbott Park Road, 60064-6125, Abbott Park, IL, USA aut Brent Putman C. Neuroscience Research, Global Pharmaceutical Research and Development, Abbott Laboratories, Dept R4PM, Bldg. AP9A, 100 Abbott Park Road, 60064-6125, Abbott Park, IL, USA aut Martin Ruth Neuroscience Research, Global Pharmaceutical Research and Development, Abbott Laboratories, Dept R4PM, Bldg. AP9A, 100 Abbott Park Road, 60064-6125, Abbott Park, IL, USA aut Surowy Carol Neuroscience Research, Global Pharmaceutical Research and Development, Abbott Laboratories, Dept R4PM, Bldg. AP9A, 100 Abbott Park Road, 60064-6125, Abbott Park, IL, USA aut Jarvis Michael Neuroscience Research, Global Pharmaceutical Research and Development, Abbott Laboratories, Dept R4PM, Bldg. AP9A, 100 Abbott Park Road, 60064-6125, Abbott Park, IL, USA aut Scott Victoria Neuroscience Research, Global Pharmaceutical Research and Development, Abbott Laboratories, Dept R4PM, Bldg. AP9A, 100 Abbott Park Road, 60064-6125, Abbott Park, IL, USA aut Inflammation Research SP Birkhäuser Verlag Basel 60/7(2011-07-01), 683-693 1023-3830 60:7<683 2011 60 11 https://doi.org/10.1007/s00011-011-0322-9 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00011-011-0322-9 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Vortherms Timothy Neuroscience Research, Global Pharmaceutical Research and Development, Abbott Laboratories, Dept R4PM, Bldg. AP9A, 100 Abbott Park Road, 60064-6125, Abbott Park, IL, USA aut NATIONALLICENCE 700 1- Swensen Andrew Neuroscience Research, Global Pharmaceutical Research and Development, Abbott Laboratories, Dept R4PM, Bldg. AP9A, 100 Abbott Park Road, 60064-6125, Abbott Park, IL, USA aut NATIONALLICENCE 700 1- Niforatos Wende Neuroscience Research, Global Pharmaceutical Research and Development, Abbott Laboratories, Dept R4PM, Bldg. AP9A, 100 Abbott Park Road, 60064-6125, Abbott Park, IL, USA aut NATIONALLICENCE 700 1- Limberis James Neuroscience Research, Global Pharmaceutical Research and Development, Abbott Laboratories, Dept R4PM, Bldg. AP9A, 100 Abbott Park Road, 60064-6125, Abbott Park, IL, USA aut NATIONALLICENCE 700 1- Neelands Torben Neuroscience Research, Global Pharmaceutical Research and Development, Abbott Laboratories, Dept R4PM, Bldg. AP9A, 100 Abbott Park Road, 60064-6125, Abbott Park, IL, USA aut NATIONALLICENCE 700 1- Janis Richard Neuroscience Research, Global Pharmaceutical Research and Development, Abbott Laboratories, Dept R4PM, Bldg. AP9A, 100 Abbott Park Road, 60064-6125, Abbott Park, IL, USA aut NATIONALLICENCE 700 1- Thimmapaya Rama Neuroscience Research, Global Pharmaceutical Research and Development, Abbott Laboratories, Dept R4PM, Bldg. AP9A, 100 Abbott Park Road, 60064-6125, Abbott Park, IL, USA aut NATIONALLICENCE 700 1- Donnelly-Roberts Diana Neuroscience Research, Global Pharmaceutical Research and Development, Abbott Laboratories, Dept R4PM, Bldg. AP9A, 100 Abbott Park Road, 60064-6125, Abbott Park, IL, USA aut NATIONALLICENCE 700 1- Namovic Marian Neuroscience Research, Global Pharmaceutical Research and Development, Abbott Laboratories, Dept R4PM, Bldg. AP9A, 100 Abbott Park Road, 60064-6125, Abbott Park, IL, USA aut NATIONALLICENCE 700 1- Zhang Di Neuroscience Research, Global Pharmaceutical Research and Development, Abbott Laboratories, Dept R4PM, Bldg. AP9A, 100 Abbott Park Road, 60064-6125, Abbott Park, IL, USA aut NATIONALLICENCE 700 1- Brent Putman C. Neuroscience Research, Global Pharmaceutical Research and Development, Abbott Laboratories, Dept R4PM, Bldg. AP9A, 100 Abbott Park Road, 60064-6125, Abbott Park, IL, USA aut NATIONALLICENCE 700 1- Martin Ruth Neuroscience Research, Global Pharmaceutical Research and Development, Abbott Laboratories, Dept R4PM, Bldg. AP9A, 100 Abbott Park Road, 60064-6125, Abbott Park, IL, USA aut NATIONALLICENCE 700 1- Surowy Carol Neuroscience Research, Global Pharmaceutical Research and Development, Abbott Laboratories, Dept R4PM, Bldg. AP9A, 100 Abbott Park Road, 60064-6125, Abbott Park, IL, USA aut NATIONALLICENCE 700 1- Jarvis Michael Neuroscience Research, Global Pharmaceutical Research and Development, Abbott Laboratories, Dept R4PM, Bldg. AP9A, 100 Abbott Park Road, 60064-6125, Abbott Park, IL, USA aut NATIONALLICENCE 700 1- Scott Victoria Neuroscience Research, Global Pharmaceutical Research and Development, Abbott Laboratories, Dept R4PM, Bldg. AP9A, 100 Abbott Park Road, 60064-6125, Abbott Park, IL, USA aut NATIONALLICENCE 773 0- Inflammation Research SP Birkhäuser Verlag Basel 60/7(2011-07-01), 683-693 1023-3830 60:7<683 2011 60 11 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer