Marcus D C
Am J Physiol. 1986 Nov;251(5 Pt 1):C662-70. doi: 10.1152/ajpcell.1986.251.5.C662.
Transepithelial electrical potential difference (VT) was measured across the vestibular labyrinth of the inner ear in vitro by puncturing the epithelial wall of the utricle with a glass microelectrode. A region of nonsensory cells of the utricle was isolated from the sensory regions by introducing columns of liquid Sylgard 184. Under control conditions, the VT of this region was +7.5 +/- 0.3 mV (means +/- SE), lumen positive. This potential difference was rapidly reduced by either 1 mM ouabain, 10-100 microM bumetanide, 0.5-5.0 mM Ba (in the bathing solution), or cooling, but not by the disulfonic stilbene, 4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid. Changes in VT due to reductions of Cl or Na or to increases of K in the bathing solution in exchange for presumably impermeant ions were observed in this region and were compared with those in a preparation in which the insulating seals were absent. The K-induced voltage change was significantly higher in the unblocked preparation, a finding consistent with a high K permeability of the sensory cells. The voltage change due to reduction of Cl was not inhibited by Cl channel blockers (9-anthracenecarboxylate and diphenylamine-2-carboxylate) in the bathing solution. These results represent the first direct demonstration that the nonsensory cells of the utricle produce a lumen-positive active-transport potential and characterize some of the properties of the cell membranes in terms of their pharmacological sensitivities and net voltage responses to changes in the bathing medium ions Na, K, and Cl.
通过用玻璃微电极穿刺椭圆囊的上皮壁,在体外测量跨内耳前庭迷路的跨上皮电势差(VT)。通过引入液态Sylgard 184柱,将椭圆囊的非感觉细胞区域与感觉区域隔离开。在对照条件下,该区域的VT为+7.5±0.3 mV(平均值±标准误),管腔呈正电位。1 mM哇巴因、10 - 100 μM布美他尼、0.5 - 5.0 mM Ba(在浴液中)或降温均可迅速降低该电势差,但二磺酸芪(4 - 乙酰氨基 - 4'-异硫氰酸芪 - 2,2'-二磺酸)则无此作用。在该区域观察到由于浴液中Cl或Na减少或K增加以置换可能不可渗透的离子而导致的VT变化,并与未形成绝缘封接的制剂中的变化进行了比较。在未封接的制剂中,K诱导的电压变化明显更高,这一发现与感觉细胞的高K通透性一致。浴液中Cl通道阻滞剂(9 - 蒽羧酸和二苯胺 - 2 - 羧酸)并未抑制由于Cl减少而引起的电压变化。这些结果首次直接证明了椭圆囊的非感觉细胞产生管腔正性的主动转运电位,并根据其药理学敏感性和对浴液介质中离子Na、K和Cl变化的净电压反应,对细胞膜的一些特性进行了表征。