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从成年豚鼠心脏房室结分离出的单细胞的特征

Characteristics of single cells isolated from the atrioventricular node of the adult guinea-pig heart.

作者信息

Yuill Kathryn H, Hancox Jules C

机构信息

Department of Physiology and Cardiovascular Research Laboratories, School of Medical Sciences, University Walk, Britsol, UK.

出版信息

Pflugers Arch. 2002 Dec;445(3):311-20. doi: 10.1007/s00424-002-0932-8. Epub 2002 Sep 26.

Abstract

To date, data regarding the cellular electrophysiology of the atrioventricular node (AVN) have derived from AVN cells isolated from the rabbit heart. The aim of this study was to characterise for the first time the electrophysiological properties of single cells isolated from the AVN of the guinea-pig heart. Cells were isolated from the AVN region by a combination of enzymatic and mechanical dispersion. Mean (+/-SEM) cell dimensions were 110.8+/-2.3 microm in length by 11.4+/-1.3 micro m in width (n=76 cells). Electrophysiological recordings were made using the whole-cell patch-clamp technique at 37 degrees C. Mean cell capacitance was 25.1+/-0.9 pF (n=43) and mean cell input resistance was 1,377+/-178 Muomega (n=21). Spontaneously active cells exhibited action potentials characteristic of cardiac pacemaker tissue. Under whole-cell voltage clamp, the mean 'zero current' potential was -39.7+/-4.1 mV (n=21). Voltage commands of 500 ms duration to a range of test potentials from a holding potential of -40 mV revealed a number of distinct current components. At potentials positive to -40 mV an early inward current was observed that exhibited a bell-shaped current voltage (I-V) relation, typical of L-type calcium current. A delayed outward current that increased progressively with test potential magnitude was also observed. Analysis of the outward current 'tails' on repolarisation to -40 mV showed this to be comprised of two components with half-maximal activation voltages of -17.2+/-1.8 mV, and +27.1+/-3.6 mV (n=13). 'Transient outward' current appeared absent from guinea-pig AVN cells. Hyperpolarising test pulses activated net inward current, for which three components could be observed: a barium-sensitive (100 microM Ba(2+)) inwardly rectifying current evident at the start of the voltage command and prominent at potentials negative to the diastolic potential range; a time-dependent, hyperpolarisation-activated current, and a residual background current. On repolarisation to -40 mV, a large inward current spike typical of cardiac Na current was observed in some cells. Notably, the following electrophysiological characteristics of guinea-pig AVN cells are distinct from those characteristics previously reported for the rabbit AVN: (1) an absence of transient outward current, (2) the presence of two components of delayed outward current, and (3) the presence of a Ba(2+)-sensitive inwardly rectifying current at negative voltages. The guinea-pig AVN isolated cell preparation may be valuable in providing additional insights into the cellular electrophysiology of this important region of the heart.

摘要

迄今为止,有关房室结(AVN)细胞电生理学的数据均来自从兔心脏分离出的AVN细胞。本研究的目的是首次对从豚鼠心脏AVN分离出的单细胞的电生理特性进行表征。通过酶解和机械分散相结合的方法从AVN区域分离细胞。平均(±标准误)细胞尺寸为长110.8±2.3微米,宽11.4±1.3微米(n = 76个细胞)。在37℃下使用全细胞膜片钳技术进行电生理记录。平均细胞电容为25.1±0.9 pF(n = 43),平均细胞输入电阻为1377±178 MΩ(n = 21)。自发活动的细胞表现出心脏起搏组织特有的动作电位。在全细胞电压钳制下,平均“零电流”电位为-39.7±4.1 mV(n = 21)。从-40 mV的保持电位向一系列测试电位施加持续500 ms的电压指令,揭示了许多不同的电流成分。在高于-40 mV的电位下,观察到一个早期内向电流,其呈现出典型的L型钙电流的钟形电流-电压(I-V)关系。还观察到一个延迟外向电流,其随着测试电位幅度的增加而逐渐增加。对复极化至-40 mV时的外向电流“尾电流”分析表明,它由两个成分组成,半最大激活电压分别为-17.2±1.8 mV和+27.1±3.6 mV(n = 13)。豚鼠AVN细胞似乎不存在“瞬时外向”电流。超极化测试脉冲激活净内向电流,可观察到三个成分:一种钡敏感(100 μM Ba²⁺)内向整流电流,在电压指令开始时明显,在低于舒张电位范围的电位时突出;一种时间依赖性、超极化激活电流,以及一种残余背景电流。在复极化至-40 mV时,在一些细胞中观察到典型的心脏钠电流的大内向电流尖峰。值得注意的是,豚鼠AVN细胞的以下电生理特性与先前报道的兔AVN的特性不同:(1)不存在瞬时外向电流,(2)存在延迟外向电流的两个成分,(3)在负电压下存在钡敏感内向整流电流。豚鼠AVN分离细胞制剂可能有助于为深入了解心脏这一重要区域的细胞电生理学提供更多见解。

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