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Related Experiment Videos

Apical Cl- channels in A6 cells

W Zeiske1, F Atia, W Van Driessche

  • 1Laboratory of Physiology, K.U. Leuven, Campus Gasthuisberg, B-3000 Leuven, Belgium.

The Journal of Membrane Biology
|December 8, 1998
PubMed
Summary
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Stimulating Xenopus A6 cells with agents increasing cyclic AMP (cAMP) activated chloride secretion and apical chloride channels. This suggests cAMP signaling may involve exocytosis of chloride channels.

Area of Science:

  • * Cell biology
  • * Physiology
  • * Molecular biology

Background:

  • * Polarized epithelial cells, specifically from the distal nephron of Xenopus laevis (A6 cell line), are crucial for ion transport.
  • * Understanding the mechanisms regulating ion secretion, particularly chloride (Cl-), is vital for comprehending kidney function.

Purpose of the Study:

  • * To investigate the role of cyclic AMP (cAMP) in regulating electrogenic chloride secretion in Xenopus A6 cells.
  • * To analyze the electrical properties, including short-circuit current (Isc), transepithelial conductance (Gt), and capacitance (CT), in response to various stimuli.
  • * To characterize the noise properties associated with ion channel activity and explore potential mechanisms of channel regulation.

Main Methods:

  • * Measurement of short-circuit current (Isc), transepithelial conductance (Gt), and electrical capacitance (CT) in polarized A6 epithelial cells.

Related Experiment Videos

  • * Apical and basolateral perfusions with specific ionic solutions (Na+- and Cl--free apical, NaCl-Ringer basolateral).
  • * Application of cAMP-increasing agents (hormones, forskolin, phosphodiesterase inhibitors) and analysis of associated electrical noise.

    • Main Results:

    • * Agents increasing cellular cAMP evoked significant increases in Gt, CT, and Isc, accompanied by Lorentzian Isc-noise, indicative of active Cl- secretion.
    • * Hormonal stimulation (arginine-vasotocin, oxytocin) resulted in a peak-plateau response, while forskolin and phosphodiesterase inhibitors showed different response patterns.
    • * All stimulants induced Lorentzian noise with a consistent corner frequency (40-50 Hz), suggesting activation of apical Cl- channels. cAMP activation could overcome hormone-induced refractoriness, and increased CT correlated with Gt, suggesting exocytosis of Cl- channels.

    Conclusions:

    • * Activation of the cAMP-messenger pathway in Xenopus A6 cells stimulates electrogenic chloride secretion.
    • * The observed increase in apical membrane capacitance (CT) suggests that cAMP signaling may involve the exocytosis of a vesicular pool of chloride channels.
    • * Noise analysis provides insights into the kinetics and characteristics of apical chloride channels, with flufenamate acting as a blocker but not a simple open-channel inhibitor.