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Related Experiment Videos

Stilbenes stimulate T84 Cl- secretion by elevating Ca2+

D J Brayden1, M E Krouse, T Law

  • 1Cystic Fibrosis Research Laboratory, Stanford University, California 94305-2130.

The American Journal of Physiology
|February 1, 1993
PubMed
Summary

4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS) and similar compounds transiently increase chloride secretion in T84 cells, likely via calcium elevation. This effect differs between confluent and nonconfluent cells, suggesting distinct calcium-activated conductances.

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Area of Science:

  • Cell Biology
  • Physiology
  • Ion Transport

Background:

  • T84 cell monolayers are a model for studying epithelial ion transport.
  • Chloride secretion is a critical process in epithelial tissues.
  • Calcium signaling plays a role in regulating ion channel activity.

Purpose of the Study:

  • To investigate the effects of 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS) and related compounds on chloride secretion in T84 cells.
  • To elucidate the role of intracellular calcium in mediating these effects.
  • To compare the mechanisms of ion transport in confluent versus nonconfluent T84 cells.

Main Methods:

  • Application of DIDS and SITS to T84 cell monolayers.
  • Measurement of short-circuit current (Isc) to assess ion transport.

Related Experiment Videos

  • Use of Fura-2 to measure intracellular calcium levels.
  • Whole-cell patch-clamp recordings to evaluate ion channel activity.

    • Main Results:

    • Basolateral DIDS and SITS induced a transient increase in Isc, followed by inhibition.
    • The DIDS-induced increase in Isc mimicked responses to thapsigargin, suggesting calcium involvement.
    • Responses were blocked by bumetanide and dependent on basolateral calcium.
    • Synergistic effects were observed with cAMP-elevating agents.
    • In nonconfluent cells, DIDS stimulated chloride conductance, unlike in confluent monolayers.

    Conclusions:

    • DIDS and SITS activate chloride secretion in T84 cells, primarily through calcium-dependent mechanisms.
    • Confluent T84 monolayers exhibit distinct calcium-activated conductances compared to nonconfluent cells.
    • The findings support a model where cyclic nucleotides activate apical chloride conductance and calcium activates basolateral potassium channels in confluent monolayers.