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A microfluidic platform for measuring electrical activity across cells.

Cédric Bathany¹, Derek L Beahm, Steve Besch

¹Department of Mechanical and Aerospace Engineering, SUNY-Buffalo, Buffalo, New York 14260, USA.

Biomicrofluidics

|September 25, 2013

Summary

This summary is machine-generated.

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This study introduces a microfluidic chip for measuring electrical conductance via gap junction channels. The novel chip demonstrates that 2-APB drug reversibly inhibits connexin 43 (Cx43) gap junction conductivity in a dose-dependent manner.

Area of Science:

Biomedical Engineering
Cell Biology
Biophysics

Background:

Gap junctions are crucial for intercellular communication, mediating direct cell-to-cell passage of ions and small molecules.
Dysfunctional gap junctions are implicated in various diseases, necessitating tools to study their activity and develop therapeutic interventions.
Connexin 43 (Cx43) is a key gap junction protein, and understanding its regulation is vital for cellular signaling research.

Purpose of the Study:

To develop and validate a novel microfluidic chip for quantifying electrical conductance across 2D cell sheets via gap junction channels.
To investigate the effects of the gap junction inhibitor 2-aminoethoxydiphenyl borate (2-APB) on the electrical coupling of connexin 43 (Cx43) gap junction channels.
To simultaneously monitor changes in electrical conductance and dye diffusion to confirm the mechanism of action of gap junction modulators.

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Main Methods:

Development of a tri-stream laminar flow microfluidic chip to create a non-conductive sucrose gap, forcing current through cell-based gap junctions.
Application of the microfluidic chip to NRK-49F cells expressing Cx43 gap junction channels.
Real-time measurement of electrical conductance and fluorescent dye diffusion in response to varying concentrations of 2-APB.

Main Results:

The microfluidic chip successfully measured electrical conductance through Cx43 gap junction channels in a 2D cell sheet.
The gap junction inhibitor 2-APB demonstrated a reversible, dose-dependent blockade of electrical conductivity.
Simultaneous measurements showed that 2-APB affected both electrical conductance and dye diffusion, confirming its action on gap junction activity.

Conclusions:

The developed microfluidic chip serves as a versatile platform for investigating gap junction properties.
The study confirms 2-APB's inhibitory effect on Cx43 gap junction channels, impacting both electrical coupling and molecular diffusion.
This technology offers a valuable tool for high-throughput screening of drugs targeting gap junction transmission for therapeutic purposes.