Ionic conductive properties of rabbit proximal straight tubule basolateral membrane

Paul A. Welling, Roger G. O'Neil

Research output: Contribution to journalArticle

Abstract

The ionic conductive properties of the nonperfused rabbit proximal straight tubule (S2) basolateral membrane were assessed by microelectrode techniques. The response of the basolateral membrane electrical potential difference, Vbl, to rapid changes in the peritubular bath concentration of K, HCO3, Na, and Cl were monitored with microelectrodes. The control steady-state Vbl averaged -41 mV (cell negative). An increase in peritubular bathing medium K concentration from 5 to 40 mM resulted in an instantaneous and sustained depolarization of +14.6 mV (27% of ΔEK). Addition of barium (2 mM) depolarized the Vbl by +15.8 mV and abolished the Vbl response to the high-K medium. In other studies, reduction of peritubular bicarbonate at constant pH from 25 to 2.5 mM instantaneously and transiently depolarized Vbl by +15.8 mV (26% of ΔEHCO3). In these same tubules reduction of peritubular Na from 126 to 2.2 mM resulted in an instantaneous and paradoxical depolarization of Vbl of +21.5 mV. Both depolarization transients resulting from reduction of Na and HCO3 were simultaneously inhibited by the addition of 4-acetamido-4′-isothiocyanostilbene-2,2′-disulfonic acid (SITS; 0.5 mM), consistent with the presence of a SITS-sensitive Na-HCO3-coupled conductive pathway. In the absence of the bicarbonate buffer, reduction of Na resulted in a small sustained hyperpolarization of -5.8 mV (5% of ΔENa). Reduction of peritubular Cl from 120 to 4 mM resulted in an instantaneous and sustained depolarization of Vbl of +5.3 mV (6% of ECl) and was not affected by the addition of bumetanide (0.1 mM). It is concluded that the basolateral membrane of the nonperfused proximal straight tubule is characterized by a major barium-sensitive K conductance and a SITS-sensitive Na-coupled HCO3 conductance that carries net negative charge. These pathways are paralleled by relatively minor, but important, Na-conductive and Cl-conductive pathways.

Original languageEnglish (US)
Pages (from-to)F940-F950
JournalAmerican Journal of Physiology - Renal Fluid and Electrolyte Physiology
Volume258
Issue number4 27-4
StatePublished - Apr 1 1990
Externally publishedYes

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4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid
Microelectrodes
Barium
Bicarbonates
Rabbits
Membranes
Bumetanide
Baths
Membrane Potentials
Buffers
Acids

Keywords

  • 4-acetamido-4′-isothiocyanostilbene-2,2′-disulfonic acid
  • Barium
  • Bumetanide
  • Chloride conductance
  • Epithelial ion transport
  • Potassium chloride cotransport
  • Potassium conductance
  • Sodium conductance
  • Sodium-dependent bicarbonate co-transport

ASJC Scopus subject areas

  • Physiology

Cite this

Ionic conductive properties of rabbit proximal straight tubule basolateral membrane. / Welling, Paul A.; O'Neil, Roger G.

In: American Journal of Physiology - Renal Fluid and Electrolyte Physiology, Vol. 258, No. 4 27-4, 01.04.1990, p. F940-F950.

Research output: Contribution to journalArticle

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N2 - The ionic conductive properties of the nonperfused rabbit proximal straight tubule (S2) basolateral membrane were assessed by microelectrode techniques. The response of the basolateral membrane electrical potential difference, Vbl, to rapid changes in the peritubular bath concentration of K, HCO3, Na, and Cl were monitored with microelectrodes. The control steady-state Vbl averaged -41 mV (cell negative). An increase in peritubular bathing medium K concentration from 5 to 40 mM resulted in an instantaneous and sustained depolarization of +14.6 mV (27% of ΔEK). Addition of barium (2 mM) depolarized the Vbl by +15.8 mV and abolished the Vbl response to the high-K medium. In other studies, reduction of peritubular bicarbonate at constant pH from 25 to 2.5 mM instantaneously and transiently depolarized Vbl by +15.8 mV (26% of ΔEHCO3). In these same tubules reduction of peritubular Na from 126 to 2.2 mM resulted in an instantaneous and paradoxical depolarization of Vbl of +21.5 mV. Both depolarization transients resulting from reduction of Na and HCO3 were simultaneously inhibited by the addition of 4-acetamido-4′-isothiocyanostilbene-2,2′-disulfonic acid (SITS; 0.5 mM), consistent with the presence of a SITS-sensitive Na-HCO3-coupled conductive pathway. In the absence of the bicarbonate buffer, reduction of Na resulted in a small sustained hyperpolarization of -5.8 mV (5% of ΔENa). Reduction of peritubular Cl from 120 to 4 mM resulted in an instantaneous and sustained depolarization of Vbl of +5.3 mV (6% of ECl) and was not affected by the addition of bumetanide (0.1 mM). It is concluded that the basolateral membrane of the nonperfused proximal straight tubule is characterized by a major barium-sensitive K conductance and a SITS-sensitive Na-coupled HCO3 conductance that carries net negative charge. These pathways are paralleled by relatively minor, but important, Na-conductive and Cl-conductive pathways.

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KW - 4-acetamido-4′-isothiocyanostilbene-2,2′-disulfonic acid

KW - Barium

KW - Bumetanide

KW - Chloride conductance

KW - Epithelial ion transport

KW - Potassium chloride cotransport

KW - Potassium conductance

KW - Sodium conductance

KW - Sodium-dependent bicarbonate co-transport

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