Caldwell Bryan J, Legrice Ian J, Hooks Darren A, Tai Dean C-S, Pullan Andrew J, Smaill Bruce H
Bioengineering Institute and Physiology Department, University of Auckland, Auckland, New Zealand.
J Cardiovasc Electrophysiol. 2005 Sep;16(9):1001-10. doi: 10.1111/j.1540-8167.2005.40558.x.
Transmembrane potentials can be recorded at multiple intramural sites in the intact heart using fiber optic probes or optrodes. The technique has considerable potential utility for studies of arrhythmia and defibrillation, but has not been validated in large mammalian hearts.
An optrode was used to acquire intramural transmembrane potentials in six isolated Langendorff-perfused pig hearts. Mechanical activity was suppressed with 2,3-butanedione monoxime (BDM). Excitation light (488 nm) was delivered to and fluorescence collected from six sites, each spaced 1.4 mm apart across the left ventricle (LV) free wall that was stained with di-4 ANEPPS. Intramural membrane potentials were compared with extracellular potentials recorded at adjacent locations in sinus rhythm, and during atrial and subepicardial ventricular pacing (1-3 Hz). In three hearts, epicardial intracellular potentials were also measured close to the optrode. Optical action potentials were reproducible, with no significant transmural variation in morphology. There was close correspondence between subepicardial optical and intracellular potentials (R2 = 0.948, n = 23). The onset of activation at and its progression across adjacent optical and extracellular recording sites were consistent, as was the variation of action potential duration (APD) with cycle length. However, there was greater variability in absolute APD estimated from optical and extracellular records (R2 = 0.773, n = 258). Comparison of extracellular potentials at the same intramural sites in vivo confirms that heart isolation and BDM slow electrical propagation and depress restitution relationships, but otherwise preserve intramural patterns of electrical activation.
We have demonstrated that our optrode provides reliable intramural transmembrane potential recordings in the isolated pig heart preparation.
使用光纤探头或光电极可以在完整心脏的多个壁内位点记录跨膜电位。该技术在心律失常和除颤研究中具有相当大的潜在应用价值,但尚未在大型哺乳动物心脏中得到验证。
使用光电极在六个离体的Langendorff灌注猪心脏中获取壁内跨膜电位。用2,3-丁二酮一肟(BDM)抑制机械活动。激发光(488nm)照射到左心室(LV)游离壁上相隔1.4mm的六个位点,并从这些位点收集荧光,左心室游离壁用di-4 ANEPPS染色。将壁内膜电位与在窦性心律、心房和心外膜下心室起搏(1-3Hz)时相邻位置记录的细胞外电位进行比较。在三个心脏中,还在靠近光电极的位置测量了心外膜细胞内电位。光学动作电位具有可重复性,形态上没有明显的跨壁变化。心外膜下光学电位和细胞内电位之间有密切的对应关系(R2 = 0.948,n = 23)。相邻光学和细胞外记录位点的激活起始及其进展是一致的,动作电位持续时间(APD)随心动周期长度的变化也是一致的。然而,从光学和细胞外记录估计的绝对APD存在更大的变异性(R2 = 0.773,n = 258)。体内相同壁内位点的细胞外电位比较证实,心脏分离和BDM会减慢电传播并抑制恢复关系,但在其他方面保留壁内电激活模式。
我们已经证明,我们的光电极在离体猪心脏制备中提供可靠的壁内跨膜电位记录。