Hypoxia augments the calcium-activated chloride current carried by anoctamin-1 in cardiac vascular endothelial cells of neonatal mice

• Type of Material: Article
• Authors: Ming-Ming Wu, Harbin Medical University; Jie Lou, Harbin Medical University; Bin-Lin Song, Harbin Medical University; Yuan-Feng Gong, Harbin Medical University; Yan-Chao Li, Harbin Medical University; Chang-Jiang Yu, Harbin Medical University; Qiu-Shi Wang, Harbin Medical University; Tian-Xing Ma, Harbin Medical University; Ke Ma, Harbin Medical University; Harrison Hartzell Jr., Emory University; Dayue Darrel Duan, University of Nevada; Dan Zhao, Harbin Medical University; Zhi-Ren Zhang, Harbin Medical University
• Language: English
• Date: 2014-07-17
• Publisher: Wiley: 12 months
• Publication Version: Final Published Version
• Copyright Statement: © 2014 The Authors. British Journal of Pharmacology published by John Wiley & Sons Ltd on behalf of The British Pharmacological Society. This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.
• License: Creative Commons Attribution-NonCommerical-NoDerivs 3.0 Unported
• Final Published Version (URL): http://dx.doi.org/10.1111/bph.12730
• Title of Journal or Parent Work: British Journal of Pharmacology
• ISSN: 0007-1188
• Volume: 171
• Issue: 15
• Start Page: 3680
• End Page: 3692
• Grant/Funding Information: This study was supported by Key Project of Chinese National Program for Fundamental Research and Development (973 Program 2014CB542401, 2012CB517803 to Z. Z.), National Natural Science Foundation of China (30871007, 81270340 and 81320108002 to Z. Z.), Doctoral Tutor Foundation of Ministry of Education (20122307110008 to Z. Z.), Overseas Talent Foundation of Department of Education, Heilongjiang Province (1154HZ11 to Z. Z.) and the Natural Science Foundation of Heilongjiang Province (ZD200807-01, ZD200807-02 to Z. Z. and QC2010097 to D. Z.).; This study was also supported by American Heart Association Western States Affiliate Grant-in-Aid (11GRNT7610161 to D. D.) and the National Institute of Health Grant (HL106256 to D. D.).
• Keywords: HYPERTENSION; EXPRESSION; Science & Technology; HISTAMINE; PHARMACOLOGY & PHARMACY; Pharmacology & Pharmacy; CONTRIBUTES; Life Sciences & Biomedicine; CA2+-ACTIVATED CL-CHANNEL; ARRIVE GUIDELINES; TMEM16A; PROLIFERATION; CA2+ ENTRY; SMOOTH-MUSCLE-CELLS
• Research Categories: Biology, Cell; Health Sciences, Pharmacy

Abstract

Background and Purpose The molecular identity of calcium-activated chloride channels (CaCCs) in vascular endothelial cells remains unknown. This study sought to identify whether anoctamin-1 (Ano1, also known as TMEM16A) functions as a CaCC and whether hypoxia alters the biophysical properties of Ano1 in mouse cardiac vascular endothelial cells (CVECs). Experimental Approach Western blot, quantitative real-time PCR, confocal imaging analysis and patch-clamp analysis combined with pharmacological approaches were used to determine whether Ano1 was expressed and functioned as CaCC in CVECs. Key Results Ano1 was expressed in CVECs. The biophysical properties of the current generated in the CVECs, including the Ca2+ and voltage dependence, outward rectification, anion selectivity and the pharmacological profile, are similar to those described for CaCCs. The density of ICl(Ca) detected in CVECs was significantly inhibited by T16A inh-A01, an Ano1 inhibitor, and a pore-targeting, specific anti-Ano1 antibody, and was markedly decreased in Ano1 gene knockdown CVECs. The density of ICl(Ca) was significantly potentiated in CVECs exposed to hypoxia, and this hypoxia-induced increase in the density of ICl(Ca) was inhibited by T16Ainh-A01 or anti-Ano1 antibody. Hypoxia also increased the current density of ICl(Ca) in Ano1 gene knockdown CVECs. Conclusions and Implications Ano1 formed CaCC in CVECs of neonatal mice. Hypoxia enhances Ano1-mediated ICl(Ca) density via increasing its expression, altering the ratio of its splicing variants, sensitivity to membrane voltage and to Ca 2+. Ano1 may play a role in the pathophysiological processes during ischaemia in heart, and therefore, Ano1 might be a potential therapeutic target to prevent ischaemic damage.