Department of Pharmacology, School of Medicine, UPR, San Juan, PR 00936-5067.
Mol Cell Biochem. 2010 Oct;343(1-2):107-13. doi: 10.1007/s11010-010-0504-8. Epub 2010 May 30.
The influence of cell swelling on cell communication was investigated in cardiomyocytes isolated from the ventricle of adult rats. Measurements of dye coupling were performed in cell pairs using intracellular dialysis of Lucifer Yellow CH. The pipette was attached to one cell of the pair and after a gig ohm seal was achieved, the membrane was ruptured by a brief suction allowing the dye to diffuse from the pipette into the cell. Fluorescence of the dye in the injected as well as in non-dialyzed cell of the pair was continuously monitored. The results indicate that in cell pairs exposed to hypotonic solution the cell volume was increased by about 60% within 35 min and the dye coupling was significantly reduced by cell swelling. Calculation of gap junction permeability (P(j)) assuming an the intracellular volume accessible to intracellular diffusion of the dye as 12% of total cell volume, showed an average P(j) value of 0.16 ± 0.04 × 10(-4) cm/s (n = 35) in the control and 0.89 ± 1.1 × 10(-5) cm (n = 40) for cells exposed to hypotonic solution (P < 0.05). Similar results were found assuming intracellular volumes accessible to the dye of 20 and 30% of total cell volume, respectively. Cell swelling did not change the rate of intracellular diffusion of the dye. The results which indicate that cell volume is an important regulator of gap junction permeability, have important implications to myocardial ischemia and heart failure as well as to heart pharmacology because changes in cell volume caused by drugs and transmitters can impair cell communication with consequent generation of slow conduction and cardiac arrhythmias.
研究了成年大鼠心室分离的心肌细胞中细胞肿胀对细胞通讯的影响。使用细胞内灌流 Lucifer Yellow CH 对细胞对中的染料偶联进行了测量。将玻璃微电极连接到细胞对的一个细胞上,当达到千兆欧密封后,通过短暂抽吸破坏细胞膜,使染料从玻璃微电极扩散到细胞中。连续监测对细胞偶联的细胞内荧光。结果表明,在暴露于低渗溶液的细胞对中,细胞体积在 35 分钟内增加了约 60%,并且细胞肿胀显著降低了染料偶联。假设染料的细胞内扩散可到达细胞内体积的 12%,计算间隙连接通透性 (P(j)),结果显示,在对照条件下,平均 P(j) 值为 0.16 ± 0.04 × 10(-4) cm/s(n = 35),而在暴露于低渗溶液的细胞中,P(j) 值为 0.89 ± 1.1 × 10(-5) cm/s(n = 40)(P < 0.05)。假设染料可到达细胞内体积的 20%和 30%,分别得到类似的结果。细胞肿胀并没有改变染料的细胞内扩散速度。这些结果表明,细胞体积是间隙连接通透性的重要调节因子,对心肌缺血和心力衰竭以及心脏药理学具有重要意义,因为药物和递质引起的细胞体积变化会损害细胞通讯,导致缓慢传导和心律失常的产生。