Botchkin L M, Matthews G
Department of Neurobiology and Behavior, State University of New York, Stony Brook 11794-5230, USA.
J Cell Physiol. 1995 Aug;164(2):286-94. doi: 10.1002/jcp.1041640209.
Membrane current and [Ca]i in rabbit nonpigmented ciliary body epithelial cells (NPE cells) were monitored with combined patch-clamp and fura-2 measurements during cell swelling induced by anisosmotic conditions. In the presence of K-channel blockers, cell swelling produced an increase in membrane current, accompanied by an increase in [Ca]i. Structural changes in the cell, associated with membrane deformation, may be the cause of the increase in [Ca]i during swelling. The conductance activated by swelling was permeable to Cl: it was dependent on the Cl concentration gradient across the cell membrane, and it was blocked by the Cl-channel blockers DIDS, SITS, NPPB, and DIOA. Although swelling increased both Cl current and [Ca]i, there was no evidence that Ca was involved in the regulation of the Cl conductance. Cell swelling activated the current even when [Ca]i was strongly buffered at an elevated level (500 nM) or at a low level (approximately 0) with internal Ca-BAPTA/Cs-BAPTA mixtures. In addition, Cl conductance was unaffected when [Ca]i was increased with a Ca ionophore. There was also no evidence that cAMP participates in the regulation of the Cl conductance: swelling activation of the current occurred in the presence of cAMP inhibitor (Rp-cAMP-S) and cAMP mimic (Sp-cAMP-S). The data suggest independent involvement of Cl conductance and internal Ca in the regulation of cell volume in NPE cells.
在等渗条件诱导的细胞肿胀过程中,采用膜片钳和fura-2测量相结合的方法监测兔非色素睫状体上皮细胞(NPE细胞)的膜电流和细胞内钙离子浓度([Ca]i)。在存在钾通道阻滞剂的情况下,细胞肿胀导致膜电流增加,同时[Ca]i也增加。与膜变形相关的细胞结构变化可能是肿胀过程中[Ca]i增加的原因。肿胀激活的电导对氯离子通透:它依赖于跨细胞膜的氯离子浓度梯度,并且被氯离子通道阻滞剂DIDS、SITS、NPPB和DIOA阻断。尽管肿胀增加了氯离子电流和[Ca]i,但没有证据表明钙离子参与了氯离子电导的调节。即使细胞内钙离子浓度用内部Ca-BAPTA/Cs-BAPTA混合物在高水平(500 nM)或低水平(约0)下被强烈缓冲,细胞肿胀仍能激活电流。此外,当用钙离子载体增加[Ca]i时,氯离子电导不受影响。也没有证据表明环磷酸腺苷(cAMP)参与氯离子电导的调节:在存在cAMP抑制剂(Rp-cAMP-S)和cAMP类似物(Sp-cAMP-S)的情况下,电流仍会因肿胀而激活。这些数据表明氯离子电导和细胞内钙离子在NPE细胞体积调节中独立发挥作用。