Cell Physics Laboratory, Department of Physiology, Columbia University College of Physicians & Surgeons, 10032, New York, New York.
J Membr Biol. 1971 Mar;5(1):20-50. doi: 10.1007/BF01870824.
Substitution of extracellular Na(+) by Li(+) causes depression of junctional membrane permeability inChironomus salivary gland cells; within 3 hr, permeability falls to so low a level that neither fluorescein nor the smaller inorganic ions any longer traverse the junctional membrane in detectable amounts (uncoupling). The effect is Li-specific: if choline(+) is the Na(+) substitute, coupling is unchanged. The Li-produced uncoupling is not reversed by restitution of Na(+). Long-term exposure (>1 hr) of the cells to Ca, Mg-free medium leads also to uncoupling. This uncoupling is fully reversible by early restitution of Ca(++) or Mg(++). Coupling is maintained in the presence of either Ca(++) or Mg(++), so long as the total divalent concentration is about 12MM. The uncoupling in Ca, Mg-free medium ensues regardless of whether the main monovalent cation is Na, Li or choline.The uncouplings are accompanied by cell depolarization. Repolarization of the cells by inward current causes restoration of coupling; the junctional conductance rises again to its normal level. The effect was shown for Li-produced uncoupling, for uncoupling by prolonged absence of external Ca(++) and Mg(++), and for uncoupling produced by dinitrophenol. In all cases, the recoupling has the same features: (1) it develops rapidly upon application of the polarizing current; (2) it is cumulative; (3) it is transient, but outlasts the current; and (4) it appears not to depend on the particular ions carrying the current from the electrodes to the cell. The recoupling is due to repolarization of nonjunctional cell membrane; recoupling can be produced at zero net currernt through the junctional membrane. Recoupling takes place also as a result of chemically produced repolarization; restoration of theK gradients in uncoupled cells causes partial recoupling during the repolarization phase.An explanation of the results on coupling is proposed in terms of known mechanisms of regulation of Ca(++) flux in cells. The uncouplings are explained by actions raising the Ca(++) level in the cytoplasmic environment of the junctional membranes; the recoupling is explained by actions lowering this Ca(++) level.
细胞外钠离子被锂离子取代会导致摇蚊唾液腺细胞连接膜通透性降低;在 3 小时内,通透性降低到极低水平,以至于荧光素和较小的无机离子都不再以可检测的量穿过连接膜(解偶联)。这种效应是锂特异性的:如果用胆碱取代钠离子,耦合不变。用锂产生的解偶联不能通过恢复钠离子来逆转。细胞长期(>1 小时)暴露于无钙镁的培养基中也会导致解偶联。这种解偶联可以通过早期恢复钙(++)或镁(++)完全逆转。只要总二价浓度约为 12MM,钙(++)或镁(++)存在时保持耦合。在存在钙(++)或镁(++)的情况下,无论主要单价阳离子是钠、锂还是胆碱,耦合都能维持。无钙镁培养基中的解偶联与主要单价阳离子无关。解偶联伴随着细胞去极化。内向电流使细胞复极化导致耦合恢复;连接电导再次上升到正常水平。该效应适用于锂产生的解偶联、长时间缺乏细胞外钙(++)和镁(++)引起的解偶联以及二硝基苯酚产生的解偶联。在所有情况下,再偶联都具有相同的特征:(1)在施加极化电流时迅速发展;(2)它是累积的;(3)它是瞬态的,但持续时间超过电流;(4)它似乎不依赖于将电流从电极带到细胞的特定离子。再偶联是由于非连接细胞膜的复极化引起的;通过连接膜可以在零净电流下产生再偶联。再偶联也可以作为化学产生的复极化的结果发生;在去偶联细胞中恢复钾梯度会导致复极化过程中部分再偶联。根据细胞中钙(++)流调节的已知机制,提出了对耦合结果的解释。解偶联是由于增加了连接膜细胞质环境中的钙(++)水平的作用引起的;再偶联是由于降低了这种钙(++)水平的作用引起的。