Yao A, Spitzer K W, Ito N, Zaniboni M, Lorell B H, Barry W H
Cardiology Division, University of Utah, Salt Lake City 84132, USA.
Cell Calcium. 1997 Dec;22(6):431-8. doi: 10.1016/s0143-4160(97)90070-1.
In cardiac muscle sarcolemmal structures such as T-tubules, caveolae and negatively charged protein-polysaccharides may affect the rate of cation exchange on the external surface of the cells. To test this hypothesis, we examined the rate of external cation exchange in adult rabbit and rat ventricular myocytes using a rapid solution switcher to change the bulk external solution within 4 ms. To assess the rate of diffusion of monovalent cations, we increased [K+]o from 4.4 to 6.6 or 8.8 mM and measured the time required to achieve a stable membrane depolarization. In rat myocytes, the mean time to 90% depolarization (t90) was significantly longer than that in rabbit myocytes (137 and 64 ms, respectively) and the difference in t90 was not associated with the cell size. To assess the time course of exchange of external Ca2+, we rapidly exposed the myocytes to 0 Ca2+-2 mM EGTA solution at specific time points before action potentials or voltage clamp steps, and measured the rate of alteration of the normalized peak [Ca2+]i transient (Fluo-3) or Ca2+ current. Exposure to 0 Ca2+-2 mM EGTA solution caused a decline in the intracellular calcium transient. In rat myocytes, the rate of decline in the [Ca2+]i transient was much slower (t90 > 1500 ms, the time required for 90% decline) than for the rabbit (t90 = 295 ms). Also, the rate of decline in the Ca2+ current was prolonged in rat myocytes (t90 = 910 ms) compared with rabbit myocytes (t90 = 241 ms). These data indicate that there is a restricted space on the external surface of sarcolemma which limits diffusion of divalent cations more markedly than monovalent cations. The extent of this limitation of cation diffusion varies between species, and may have functional significance.
在心肌中,肌膜结构如横管、小凹以及带负电荷的蛋白多糖可能会影响细胞外表面阳离子交换的速率。为了验证这一假设,我们使用快速溶液切换器在4毫秒内改变细胞外总体溶液,检测成年兔和大鼠心室肌细胞的细胞外阳离子交换速率。为了评估单价阳离子的扩散速率,我们将细胞外钾离子浓度([K+]o)从4.4毫摩尔/升提高到6.6或8.8毫摩尔/升,并测量达到稳定膜去极化所需的时间。在大鼠心肌细胞中,达到90%去极化的平均时间(t90)明显长于兔心肌细胞(分别为137毫秒和64毫秒),且t90的差异与细胞大小无关。为了评估细胞外钙离子交换的时间进程,我们在动作电位或电压钳制步骤之前的特定时间点,将心肌细胞快速暴露于0钙离子-2毫摩尔/升乙二醇双四乙酸(EGTA)溶液中,并测量归一化峰值细胞内钙离子瞬变(Fluo-3)或钙离子电流的变化速率。暴露于0钙离子-2毫摩尔/升EGTA溶液会导致细胞内钙离子瞬变下降。在大鼠心肌细胞中,细胞内钙离子瞬变的下降速率比兔心肌细胞慢得多(t90>1500毫秒,即下降90%所需的时间)(兔心肌细胞t90=295毫秒)。此外,与兔心肌细胞(t90=241毫秒)相比,大鼠心肌细胞中钙离子电流的下降速率延长(t90=910毫秒)。这些数据表明,肌膜外表面存在一个受限空间,该空间对二价阳离子扩散的限制比对单价阳离子更为明显。这种阳离子扩散限制的程度在不同物种之间有所不同,并且可能具有功能意义。