Graf J, Henderson R M, Krumpholz B, Boyer J L
J Membr Biol. 1987;95(3):241-54. doi: 10.1007/BF01869486.
The basic electrical properties of an isolated rat hepatocyte couplet (IRHC) system have been analyzed using classical techniques of epithelial electrophysiology, including measurement of electric potentials, resistances and intracellular ion activities. Applications of these techniques are discussed with respect to their limitations in small isolated cells. Mean intracellular and intracanalicular membrane potentials ranged from -23.7 to -46.7 and -4.3 to -5.9 mV, respectively. Membrane resistances were determined using an equivalent circuit analysis modified according to the geometry of the IRHC system. Resistances of the sinusoidal (basolateral) and canalicular (luminal) cell membranes and tight junctions averaged 0.15 and 0.78 G omega and 25 m omega, respectively. The cells are electrically coupled via low resistance intercellular communications (approximately 58 M omega). Intracellular ion activities for Na+, K+ and Cl- averaged 12.2, 88.1 and 17.7 mmol/liter, respectively. The basolateral membrane potential reveals a permeability sequence of PK greater than PCl greater than PNa. The luminal potential showed minimal dependence on changes in transjunctional ion gradients, indicating a poor ion selectivity of the paracellular pathway. The electrogenic (Na+-K+)-ATPase contributes little to the luminal and cellular negative electric potential. Therefore, the luminal potential probably results from the secretion of impermeant ions and a Donnan distribution of permeant ions, a mechanism which provides the osmotic driving force for bile formation. By providing the unique opportunity to measure luminal potentials, this isolated hepatocyte system permits study of secretory mechanisms for the first time in a mammalian gland using electrophysiologic techniques.
已使用上皮电生理学的经典技术分析了分离的大鼠肝细胞偶联物(IRHC)系统的基本电学特性,包括测量电势、电阻和细胞内离子活性。讨论了这些技术在小的分离细胞中的应用及其局限性。平均细胞内和胆小管膜电位分别为-23.7至-46.7 mV和-4.3至-5.9 mV。使用根据IRHC系统的几何形状修改的等效电路分析来确定膜电阻。窦状(基底外侧)和胆小管(管腔)细胞膜以及紧密连接的电阻平均分别为0.15和0.78 GΩ以及25 mΩ。细胞通过低电阻细胞间通讯(约58 MΩ)电耦合。Na +、K +和Cl -的细胞内离子活性平均分别为12.2、88.1和17.7 mmol/升。基底外侧膜电位显示出PK大于PCl大于PNa的通透性顺序。管腔电位对跨连接离子梯度变化的依赖性最小,表明细胞旁途径的离子选择性较差。电生(Na + -K +)-ATP酶对管腔和细胞的负电位贡献很小。因此,管腔电位可能是由不可渗透离子的分泌和可渗透离子的唐南分布引起的,这是一种为胆汁形成提供渗透驱动力的机制。通过提供测量管腔电位的独特机会,这个分离的肝细胞系统首次允许使用电生理技术研究哺乳动物腺体中的分泌机制。