Microelectrode and impedance analysis of anion secretion in calu-3 cells

Tsutomu Tamada, Martin J. Hug, Raymond A. Frizzell, Robert J. Bridges

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25 Citations (Scopus)

Abstract

Calu-3 cells secrete HCO3- in response to cAMP agonists but can be stimulated to secrete Cl- with K+ channel activating agonists. Microelectrode and impedance analysis experiments were performed to obtain a better understanding of the conductances and driving forces involved in these different modes of anion secretion in Calu-3 cells. Microelectrode studies revealed apical and basolateral membrane depolarizations upon the addition of forskolin (Vap -52 mV vs. -21 mV; Vbl 60 mV vs. -44 mV) that paralleled the hyperpolarization of the mucosal negative transepithelial voltage (VT -8 mV vs. -23 mV). These changes were accompanied by a decrease in the apical membrane fractional resistance (FRap) from approximately 0.50 to 0.08, consistent with the activation of an apical membrane conductance. The subsequent addition of 1-ethyl-2-benzimidazolinone (1-EBIO), a K+ channel activator, hyperpolarized Vap to - 27 mV, Vbl to - 60 mV and VT to - 33 mV. Impedance analysis revealed the apical membrane resistance (Rap) of the forskolin-stimulated cells was less than 20 Ω cm2, indeed in most monolayers Rap fell to less than 5 W cm2. The impedance derived estimate of the basolateral membrane resistance (Rbl) was approximately 170 Ω cm2 in forskolin treated cells and fell to 50 Ω cm2 with the addition of 1-EBIO. Using these values for the Rbl and the FRap value of 0.08 yields a Rap of approximately 14 Ω cm2 in the presence of forskolin and 4 Ω cm2 in the presence of forskolin plus 1-EBIO. Thus, by two independent methods, forskolin-stimulated Calu-3 cells are seen to have a very high apical membrane conductance of 50 to 200 mS/cm2. Therefore, we would assert that even at one-tenth the anion selectivity for Cl-, this high conductance could support the conductive exit of HCO3- across the apical membrane. We further propose that this high apical membrane conductance serves to clamp the apical membrane potential near the equilibrium potential for Cl- and thereby provides the driving force for HCO3- secretion in forskolin-stimulated Calu-3 cells. The hyperpolarization of Vap and Vbl caused by 1-EBIO provides a driving force for Cl- exit across the apical membrane, inhibits the influx of HCO3- on the Na+:HCO3- cotransporter across the basolateral membrane, activates the basolateral membrane Na+:K:2Cl- cotransporter and thereby provides the switch from HCO3- secretion to Cl-secretion.

Original languageEnglish
Pages (from-to)219-228
Number of pages10
JournalJournal of the Pancreas
Volume2
Issue number4
Publication statusPublished - 2001 Jul
Externally publishedYes

Keywords

  • Bicarbonates
  • Cholrides
  • Secretions

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Hepatology
  • Endocrinology

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