Cheng Hongwei, Li Jue, James Andrew F, Inada Shin, Choisy Stéphanie C M, Orchard Clive H, Zhang Henggui, Boyett Mark R, Hancox Jules C
School of Physiology, Pharmacology & Neuroscience, Biomedical Sciences Building, University of Bristol, Bristol BS8 1TD, UK.
Institute of Cardiovascular Sciences, University of Manchester, Core Technology Facility, 46 Grafton Street, Manchester M13 9NT, UK.
J Mol Cell Cardiol. 2016 Aug;97:114-24. doi: 10.1016/j.yjmcc.2016.04.014. Epub 2016 Apr 27.
Background inward sodium current (IB,Na) that influences cardiac pacemaking has been comparatively under-investigated. The aim of this study was to determine for the first time the properties and role of IB,Na in cells from the heart's secondary pacemaker, the atrioventricular node (AVN). Myocytes were isolated from the AVN of adult male rabbits and mice using mechanical and enzymatic dispersion. Background current was measured using whole-cell patch clamp and monovalent ion substitution with major voltage- and time-dependent conductances inhibited. In the absence of a selective pharmacological inhibitor of IB,Na, computer modelling was used to assess the physiological contribution of IB,Na. Net background current during voltage ramps was linear, reversing close to 0mV. Switching between Tris- and Na(+)-containing extracellular solution in rabbit and mouse AVN cells revealed an inward IB,Na, with an increase in slope conductance in rabbit cells at -50mV from 0.54±0.03 to 0.91±0.05nS (mean±SEM; n=61 cells). IB,Na magnitude varied in proportion to [Na(+)]o. Other monovalent cations could substitute for Na(+) (Rb(+)>K(+)>Cs(+)>Na(+)>Li(+)). The single-channel conductance with Na(+) as charge carrier estimated from noise-analysis was 3.2±1.2pS (n=6). Ni(2+) (10mM), Gd(3+) (100μM), ruthenium red (100μM), or amiloride (1mM) produced modest reductions in IB,Na. Flufenamic acid was without significant effect, whilst La(3+) (100μM) or extracellular acidosis (pH6.3) inhibited the current by >60%. Under the conditions of our AVN cell simulations, removal of IB,Na arrested spontaneous activity and, in a simulated 1D-strand, reduced conduction velocity by ~20%. IB,Na is carried by distinct low conductance monovalent non-selective cation channels and can influence AVN spontaneous activity and conduction.
影响心脏起搏的内向背景钠电流(IB,Na)相对研究较少。本研究的目的是首次确定IB,Na在心脏次级起搏点房室结(AVN)细胞中的特性和作用。使用机械和酶分散法从成年雄性兔和小鼠的AVN中分离出心肌细胞。使用全细胞膜片钳测量背景电流,并用主要的电压和时间依赖性电导被抑制的单价离子替代法进行测量。在没有IB,Na的选择性药理抑制剂的情况下,使用计算机建模来评估IB,Na的生理贡献。电压斜坡期间的净背景电流是线性的,反转电位接近0mV。在兔和小鼠AVN细胞中,在含Tris和含Na(+)的细胞外溶液之间切换显示出内向的IB,Na,兔细胞在-50mV时斜率电导从0.54±0.03增加到0.91±0.05nS(平均值±标准误;n = 61个细胞)。IB,Na的大小与[Na(+)]o成比例变化。其他单价阳离子可以替代Na(+)(Rb(+)>K(+)>Cs(+)>Na(+)>Li(+))。通过噪声分析估计,以Na(+)作为电荷载体的单通道电导为3.2±1.2pS(n = 6)。Ni(2+)(10mM)、Gd(3+)(100μM)、钌红(100μM)或氨氯地平(1mM)使IB,Na适度降低。氟芬那酸无显著影响,而La(3+)(100μM)或细胞外酸中毒(pH6.3)使电流抑制>60%。在我们的AVN细胞模拟条件下,去除IB,Na会使自发活动停止,并且在模拟的一维链中,传导速度降低约20%。IB,Na由不同的低电导单价非选择性阳离子通道携带,并可影响AVN的自发活动和传导。