Hüser J, Wang Y G, Sheehan K A, Cifuentes F, Lipsius S L, Blatter L A
Loyola University Chicago, Stritch School of Medicine, Department of Physiology, Maywood, IL 60153, USA.
J Physiol. 2000 May 1;524 Pt 3(Pt 3):795-806. doi: 10.1111/j.1469-7793.2000.00795.x.
Electromechanical alternans was characterized in single cat atrial and ventricular myocytes by simultaneous measurements of action potentials, membrane current, cell shortening and changes in intracellular Ca2+ concentration ([Ca2+]i). Using laser scanning confocal fluorescence microscopy, alternans of electrically evoked [Ca2+]i transients revealed marked differences between atrial and ventricular myocytes. In ventricular myocytes, electrically evoked [Ca2+]i transients during alternans were spatially homogeneous. In atrial cells Ca2+ release started at subsarcolemmal peripheral regions and subsequently spread toward the centre of the myocyte. In contrast to ventricular myocytes, in atrial cells propagation of Ca2+ release from the sarcoplasmic reticulum (SR) during the small-amplitude [Ca2+]i transient was incomplete, leading to failures of excitation-contraction (EC) coupling in central regions of the cell. The mechanism underlying alternans was explored by evaluating the trigger signal for SR Ca2+ release (voltage-gated L-type Ca2+ current, ICa,L) and SR Ca2+ load during alternans. Voltage-clamp experiments revealed that peak ICa,L was not affected during alternans when measured simultaneously with changes of cell shortening. The SR Ca2+ content, evaluated by application of caffeine pulses, was identical following the small-amplitude and the large-amplitude [Ca2+]i transient. These results suggest that the primary mechanism responsible for cardiac alternans does not reside in the trigger signal for Ca2+ release and SR Ca2+ load. beta-Adrenergic stimulation with isoproterenol (isoprenaline) reversed electromechanical alternans, suggesting that under conditions of positive cardiac inotropy and enhanced efficiency of EC coupling alternans is less likely to occur. The occurrence of electromechanical alternans could be elicited by impairment of glycolysis. Inhibition of glycolytic flux by application of pyruvate, iodoacetate or beta-hydroxybutyrate induced electromechanical and [Ca2+]i transient alternans in both atrial and ventricular myocytes. The data support the conclusion that in cardiac myocytes alternans is the result of periodic alterations in the gain of EC coupling, i. e. the efficacy of a given trigger signal to release Ca2+ from the SR. It is suggested that the efficiency of EC coupling is locally controlled in the microenvironment of the SR Ca2+ release sites by mechanisms utilizing ATP, produced by glycolytic enzymes closely associated with the release channel.
通过同时测量动作电位、膜电流、细胞缩短和细胞内钙离子浓度([Ca2+]i)的变化,对单个猫心房和心室肌细胞中的机电交替进行了表征。使用激光扫描共聚焦荧光显微镜,电诱发的[Ca2+]i瞬变的交替揭示了心房和心室肌细胞之间的显著差异。在心室肌细胞中,交替期间电诱发的[Ca2+]i瞬变在空间上是均匀的。在心房细胞中,Ca2+释放始于肌膜下周边区域,随后向肌细胞中心扩散。与心室肌细胞不同,在心房细胞中,小幅度[Ca2+]i瞬变期间从肌浆网(SR)释放的Ca2+传播不完全,导致细胞中心区域兴奋-收缩(EC)偶联失败。通过评估SR Ca2+释放的触发信号(电压门控L型Ca2+电流,ICa,L)和交替期间的SR Ca2+负荷,探索了交替的潜在机制。电压钳实验表明,与细胞缩短变化同时测量时,交替期间ICa,L峰值不受影响。通过施加咖啡因脉冲评估的SR Ca2+含量,在小幅度和大幅度[Ca2+]i瞬变后是相同的。这些结果表明,心脏交替的主要机制不在于Ca2+释放的触发信号和SR Ca2+负荷。用异丙肾上腺素进行β-肾上腺素能刺激可逆转机电交替,这表明在正性心肌收缩力和增强的EC偶联效率条件下,交替不太可能发生。机电交替的发生可由糖酵解受损引起。应用丙酮酸、碘乙酸或β-羟基丁酸抑制糖酵解通量可在心房和心室肌细胞中诱导机电和[Ca2+]i瞬变交替。数据支持这样的结论,即在心肌细胞中,交替是EC偶联增益周期性改变的结果,即给定触发信号从SR释放Ca2+的功效。有人提出,EC偶联效率在SR Ca2+释放位点的微环境中通过利用ATP的机制进行局部控制,ATP由与释放通道紧密相关的糖酵解酶产生。