White J F
J Membr Biol. 1986;92(1):75-89. doi: 10.1007/BF01869017.
The characteristics of Cl- movement across luminal and basolateral membranes of Amphiuma intestinal absorptive cells were studied using Cl(-)-sensitive microelectrodes and tracer 36Cl techniques. Intracellular Cl- activity (aiCl) was unchanged when serosal Cl- was replaced; when luminal Cl- was replaced cell Cl- was rapidly lost. Accordingly, the steady state aiCl could be varied by changing the luminal [Cl]. As luminal [Cl] was raised from 1 to 86 mM, aiCl rose in a linear manner, the mucosal membrane hyperpolarized, and the transepithelial voltage became serosa negative. In contrast, the rate of Cl- transport from the cell into the serosal medium, measured as the SITS-inhibitable portion of the Cl- absorptive flux, attained a maximum when aiCl reached an apparent value of 17 mM, indicating the presence of a saturable, serosal transport step. The stilbene-insensitive absorptive flux was linear with luminal [Cl], suggestive of a paracellular route of movement. Intracellular aCl was near electrochemical equilibrium at all but the lowest values of luminal [Cl] after interference produced by other anions was taken into account. aiCl was unaffected by Na replacement, removal of medium K, or elevation of medium HCO-3. Mucosae labeled with 36Cl lost isotope into both luminal and serosal media at the same rate and from compartments of equal capacity. Lowering luminal [Cl] or addition of theophylline enhanced luminal Cl- efflux. It is concluded that a conductive Cl leak pathway is present in the luminal membrane. Serosal transfer is by a saturable, stilbene-inhibitable pathway. Luminal Cl- entry appears to be passive, but an electrogenic uptake cannot be discounted.
利用氯离子敏感微电极和示踪剂³⁶Cl技术研究了蚓螈肠吸收细胞跨腔面膜和基底外侧膜的氯离子移动特性。当浆膜氯离子被替代时,细胞内氯离子活性(aiCl)不变;当腔面膜氯离子被替代时,细胞内氯离子迅速丢失。因此,通过改变腔内[Cl]可以改变稳态aiCl。当腔内[Cl]从1 mM升高到86 mM时,aiCl呈线性上升,黏膜膜超极化,跨上皮电压变为浆膜负电位。相反,以氯离子吸收通量中SITS可抑制部分来衡量,氯离子从细胞转运到浆膜介质中的速率在aiCl达到17 mM的表观值时达到最大值,表明存在一个可饱和的浆膜转运步骤。对二苯乙烯不敏感的吸收通量与腔内[Cl]呈线性关系,提示存在细胞旁移动途径。在考虑了其他阴离子产生的干扰后,除了最低的腔内[Cl]值外,细胞内aCl在所有情况下都接近电化学平衡。aiCl不受钠离子替代、去除培养基中的钾离子或提高培养基中HCO₃⁻的影响。用³⁶Cl标记的黏膜以相同的速率且从容量相等的区室向腔内和浆膜介质中丢失同位素。降低腔内[Cl]或添加茶碱可增强腔内氯离子外流。结论是腔面膜中存在一条传导性氯离子泄漏途径。浆膜转运是通过一条可饱和的、对二苯乙烯敏感的途径。腔内氯离子进入似乎是被动的,但不能排除电生性摄取。