Abstract
Vestibular dark cell epithelium secretes K+ via I sKchannels in the apical membrane. The previous observation that disulfonic stilbenes increased the equivalent short circuit current (I sc) suggested that these agents might be useful investigative tools in this tissue. The present experiments were conducted to determine if the increase in I scwas associated with an increase in K+ flux and if the effect was directly on the I sKchannel or indirectly via a cytosolic intermediary. Measurements of transepithelial K+ flux with the K+-selective vibrating probe and of changes in net cellular solute flux by measurements of epithelial cell height showed that 4,4′-diisothiocyanatostilbene-2,2′-disulfonic acid (DIDS) increased K+ flux by a factor of 1.96±0.71 and caused net solute efflux. The apical membrane was partitioned with a macropatch pipette and DIDS was applied either to the membrane outside the pipette, inside the pipette or to the entire apical membrane. DIDS inside the pipette increased the current across the patch, the membrane conductance, the slowly-inactivating (I sK) component of the membrane current and shifted the reversal voltage toward the equilibrium potential for K+. DIDS outside the patch decreased the patch current and conductance, consistent with shunting of current away from the membrane patch. These findings strongly support the notion that DIDS increases K+ secretion through I sKchannels in the apical membrane of vestibular dark cell epithelium by acting directly on the channels or on a tightly colocalized membrane component.
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Bültmann, R., Starke, K. 1994. Blockade by 4.4′-diisothiocyanatostilbene-2.2′-disulfonate (DIDS) of P2-purinoceptors in rat vas deferens. Br. J. Pharmacol. 112:690–694
Busch, A.E., Herzer, T., Wagner, C.A., Schmidt, F., Raber, G., Waldegger, S., Lang, F. 1994. Positive regulation by chloride channel blockers of I sKchannels expressed in Xenopus oocytes. Mol. Pharmacol. 46:750–753
Cabantchik, Z.I., Greger, R. 1992. Chemical probes for anion transporters of mammalian cell membranes. Am. J. Physiol. Cell Physiol. 262:C803-C827
Cabantchik, Z.I., Rothstein, A. 1974. Membrane proteins related to anion permeability of human red blood cells. I. Localization of disulfonic stilbene binding sites in proteins involved in permeation. J. Membrane Biol. 15:207–226
Frace, A.M., Eaton, D.C. 1992. Chemical modification of Ca2+activated potassium channels of GH3 anterior pituitary cells. Am. J. Physiol. 263:C1081-C1087
Gögelein, H., Pfannmüller, B. 1989. The nonselective cation channel in the basolateral membrane of rat exocrine pancreas. Inhibition by 3′,5′-dichlorophenylamine-2-carboxylic acid (DCDPC) and activation by stilbene disulfonates. Pfluegers Arch. 413:287–298
Lapointe, J.Y., Szabo, G. 1987. A novel holder allowing internal perfusion of patch-clamp pipettes. Pfluegers Arch. 410:212–216
Liu, J., Kozakura, K., Marcus, D.C. 1995. Evidence for purinergic receptors in vestibular dark cell and strial marginal cell epithelia of gerbil. Auditory Neurosci. (in press)
Marcus, D.C. 1986. Transepithelial electrical potential of nonsensory region of gerbil utricle in vitro. Am. J. Physiol. 251:C662-C670
Marcus, D.C., Liu, J., Wangemann, P. 1994. Transepithelial voltage and resistance of vestibular dark cell epithelium from the gerbil ampulla. Hear. Res. 73:101–108
Marcus, D.C., Marcus, N.Y. 1989. Transepithelial electrical responses to Cl− of nonsensory region of gerbil utricle. Biochim. Biophys. Acta 987:56–62
Marcus, D.C., Shen, Z. 1994. Slowly activating, voltage-dependent K+ conductance is apical pathway for K+ secretion in vestibular dark cell epithelium. Am. J. Physiol. 267:C857-C864
Marcus, D.C., Shipley, A.M. 1994. Potassium secretion by vestibular dark cell epithelium demonstrated by vibrating probe. Biophys. J. 66:1939–1942
Marcus, D.C., Takeuchi, S., Wangemann, P. 1992. Ca2+-activated nonselective cation channel in apical membrane of vestibular dark cells. Am. J. Physiol. Cell Physiol. 262:C1423-C1429
Shen, Z., Liu, J., Marcus, D.C. 1994. Direct activation of a conductive pathway in the apical membrane of vestibular dark cells by a disulfonic stilbene (DIDS). Assoc. Res. Otolaryngol. 17:134 (Abstr.)
Shen, Z., Marcus, D.C. 1995. DIDS increases the I sKcurrent across the apical membrane of vestibular dark cells of gerbil. Assoc. Res. Otolaryngol. 18:99 (Abstr.)
Soejima, M., Noma, A. 1984. Mode of regulation of the AChsensitive K-channel by the muscarinic receptor in rabbit atrial cells. Pfluegers Arch. 400:424–431
Soltoff, S.P., McMillian, M.K., Talamo, B.R., Cantley, L.C. 1993. Blockade of ATP binding site of P2 purinoceptors in rat parotid acinar cells by isothiocyanate compounds. Biochem. Pharmacol. 45:1936–1940
Tabcharani, J.A., Low, W., Elie, D., Hanrahan, J.W. 1990. Lowconductance chloride channel activated by cAMP in the epithelial cell line T84. FEBS Lett. 270:157–164
Takeuchi, S., Marcus, D.C., Wangemann, P. 1992. Maxi K+ channel in apical membrane of vestibular dark cells. Am. J. Physiol. Cell Physiol. 262:C1430-C1436
Tenenhouse, H.S., Martel, J. 1993. Na+ dependent sulfate transport in opossum kidney cells is DIDS sensitive. Am. J. Physiol. Cell Physiol. 265:C54-C61
Wangemann, P., Liu, J., Marcus, D.C. 1995. Ion transport mechanisms responsible for K+ secretion and the transepithelial voltage across marginal cells of stria vascularis in vitro. Hear. Res. 84:19–29
Wangemann, P., Marcus, D.C. 1989. Membrane potential measurements of transitional cells from the crista ampullaris of the gerbil. Effects of barium, quinidine, quinine, tetraethylammonium, cesium, ammonium, thallium and ouabain. Pfluegers Arch. 414:656–662
Wangemann, P., Marcus, D.C. 1990. K+-induced swelling of vestibular dark cells is dependent on Na+ and Cl− and inhibited by piretanide. Pfluegers Arch. 416:262–269
Wangemann, P., Shiga, N., Welch, C., Marcus, D.C. 1992. Evidence for the involvement of a K+ channel in isosmotic cell shrinking in vestibular dark cells. Am. J. Physiol. 263:C616-C622
Wehner, F., Rosin-Steiner, S., Beetz, G., Sauer, H. 1993. The anion transport inhibitor DIDS increases rat hepatocyte K+ conductance via uptake through the bilirubin pathway. J. Physiol. 471:617–635
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We thank Dr. Peter J.S. Smith and Alan Shipley of the National Vibrating Probe Facility at the Marine Biological Laboratory at Woods Hole, MA for their support and assistance in the measurements of K+ flux. This work was supported by National Institutes of Health grants R01-DC00212, R29-DC1098 and P41-RR01395.
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Shen, Z., Liu, J., Marcus, D.C. et al. DIDS increases K+ secretion through an I sKchannel in apical membrane of vestibular dark cell epithelium of gerbil. J. Membarin Biol. 146, 283–291 (1995). https://doi.org/10.1007/BF00233948
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DOI: https://doi.org/10.1007/BF00233948