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Functional Analysis of Connexin Channels in Cultured Cells by Neurobiotin Injection and Visualization.
Philipp Wörsdörfer1, Klaus Willecke2
1Institute of Anatomy and Cell Biology, University of Würzburg, Koellikerstraße 6, 97070 Würzburg, Germany.
Bio-Protocol
|September 20, 2021
Summary
This study details a method for assessing cell-to-cell communication using tracer substances. It quantifies direct intercellular communication by visualizing Neurobiotin injected into cultured cells.
Area of Science:
- Cell Biology
- Biophysics
Background:
- Gap junction channels mediate direct intercellular communication between adjacent cells.
- Assessing the functionality of gap junctions is crucial for understanding cellular networks.
Purpose of the Study:
- To describe a protocol for quantifying direct intercellular communication.
- To detail the iontophoretic injection and visualization of Neurobiotin in cultured cells.
Main Methods:
- Microinjection of low molecular weight tracer substances into cultured cells.
- Iontophoretic injection of Neurobiotin.
- Visualization of tracer distribution to assess intercellular coupling.
Main Results:
- The method allows for precise quantification of the extent of direct intercellular communication.
- Neurobiotin effectively visualizes functional gap junction channels.
Conclusions:
- Microinjection of tracer substances is a reliable method for assessing gap junction function.
- This protocol provides a standardized approach for evaluating intercellular communication.
