title: Evidence that 2-aminoethoxydiphenyl borate provokes fibrillation in perfused rat hearts via voltage-independent calcium channels.
abstract: We tested whether 2-aminoethoxydiphenyl borate (2-APB) induces arrhythmia in perfused rat hearts and whether this arrhythmia might result from the activation of voltage-independent calcium channels. Rat hearts were Langendorff perfused and beat under sinus rhythm. An isovolumic balloon inserted into the left ventricle was used to record mechanical function while bipolar electrograms were recorded from electrodes sutured to the base and the apex of hearts. Western and immunofluorescence analyses were performed on rat left ventricular protein extracts and left ventricular frozen sections, respectively. Rat ventricular myocytes express Orai 1 and Orai 3, and ventricle also contains the Orai regulator Stim1. Rat hearts (n=5) perfused with Krebs-Henseleit (KH) alone maintained sinus rhythm at 4.8 ± 0.1 Hz and stable mechanical function. By contrast, perfusing hearts (n=5) with (KH+22 μM 2-APB) provoked a period of tachycardic ectopy at rates of up to 10.8 ± 0.2 Hz. As perfusion with (KH+22 μM 2-APB) continued, the rate of spontaneous ventricular depolarization increased to 21.8 ± 1.2 Hz and became disorganized. Heart mechanical function collapsed as developed pressure decreased from 87 ± 8.8 to 3.5 ± 1.9 mm Hg. Flow rate did not change between normal (16.6 ± 0.9 ml/min) and fibrillating (17.4 ± 0.8 ml/min) hearts. The addition of 20 μM 1-[2-(4-methoxyphenyl)-2-[3-(4-methoxyphenyl) propoxy]ethyl-1H-imidazole (SKF-96365) to (KH+22 μM 2-APB) perfusates (n=4) restored sinus rhythm and heart mechanical output. These data indicate that activating myocardial voltage-independent calcium channels, possibly the Orais, may be a novel cause of ventricular arrhythmia.
authors.affiliation: The Department of Medicine, The University of Alabama at Birmingham, Birmingham, AL 35294, USA.
authors.forename: Peipei
authors.initials: P
authors.lastname: Wang
authors.forename: Patrick K
authors.initials: PK
authors.lastname: Umeda
authors.forename: Oleg F
authors.initials: OF
authors.lastname: Sharifov
authors.forename: Brian A
authors.initials: BA
authors.lastname: Halloran
authors.forename: Edlue
authors.initials: E
authors.lastname: Tabengwa
authors.forename: Hernan E
authors.initials: HE
authors.lastname: Grenett
authors.forename: Ferdinand
authors.initials: F
authors.lastname: Urthaler
authors.forename: Paul E
authors.initials: PE
authors.lastname: Wolkowicz
mesh_terms: D000818:Animals; D001896:Boron Compounds / Q000633:toxicity*; D015220:Calcium Channels / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D008297:Male; D032383:Myocytes, Cardiac; D000071740:ORAI1 Protein; D051381:Rats; D017207:Rats, Sprague-Dawley; D014693:Ventricular Fibrillation / Q000139:chemically induced*
chemical_list: D001896:Boron Compounds; D015220:Calcium Channels; D000071740:ORAI1 Protein; C532493:Orai1 protein, rat; C573659:Orai3 protein, rat; C109986:2-aminoethoxydiphenyl borate