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Patch Clamp

Patch Clamp

Many fundamental cell functions such as muscle contraction and nerve transmission rely on the electrical signals produced by the movement of positively and negatively charged ions across the cell membrane. One competent method to record current flowing across the whole cell or single ion channel is the patch-clamp technique.
In this method, a glass micropipette containing electrolyte solution is tightly sealed against a small portion of the cell membrane. As a result, a patch of the cell...

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Evidence for basolateral but not apical membrane localization of outwardly rectifying depolarization-induced Cl(-)

T H Hwang¹, H J Lee, N K Lee

¹Department of Pharmacology & Science Institute of Medicine, Dong-A Medical College, Pusan, South Korea 602-103.

The Journal of Membrane Biology

|August 10, 2000

Summary

This summary is machine-generated.

This study identifies a novel depolarization-induced chloride current (BORDIC) on the basolateral membrane of airway epithelia. This finding advances our understanding of ion channel function in respiratory tissues.

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Area of Science:

Cell Biology
Physiology
Ion Transport

Background:

Primary cultured airway epithelia serve as a model for ion channel studies.
Nystatin permeabilization allows investigation of apical or basolateral membrane ion channels.

Purpose of the Study:

To investigate depolarization-induced chloride currents in airway epithelia.
To characterize the properties and localization of these currents.

Main Methods:

Utilized rat primary cultured-airway monolayers.
Employed nystatin permeabilization for selective membrane access.
Applied voltage clamping to measure ion currents.
Investigated halide permeability and sensitivity to DIDS and bumetanide.

Main Results:

Identified 4,4'-diisothiocyanatostilbene-2, 2'-disulfonic acid (DIDS)-sensitive outwardly rectifying depolarization-induced Cl(-) (BORDIC) currents on the basolateral membrane after apical permeabilization.
Observed no significant Cl(-) current on the apical membrane after basolateral permeabilization.
Determined the halide permeability sequence for BORDIC current as Br(-) = I(-) > Cl(-).
Found BORDIC current unaffected by basolateral bumetanide.
Demonstrated that basolateral DIDS inhibits CFTR-mediated short-circuit current in intact airway epithelia.

Conclusions:

This study provides the first evidence of depolarization-induced Cl(-) currents on the basolateral membrane of airway epithelia.
These findings contribute to a deeper understanding of ion transport mechanisms in the airway epithelium.