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Correlation of open cell-attached and excised patch clamp techniques
1Department of Internal Medicine, Yale School of Medicine, New Haven, CT, USA.
The Journal of Membrane Biology
|November 1, 1995
Summary
The nystatin-induced open cell-attached patch offers a more stable alternative to excised patches for studying amiloride-sensitive sodium channels. This method improves preparation success rates for ion channel research.
Area of Science:
- Biophysics
- Cell Physiology
Background:
- Excised patch clamp techniques can suffer from instability due to seal loss and rundown.
- The amiloride-sensitive Na+ channel in A6 cells is a target for single channel analysis.
Purpose of the Study:
- To evaluate the nystatin-induced open cell-attached patch as a stable alternative to excised inside-out patches.
- To compare the stability and channel characteristics of the two patch configurations.
Main Methods:
- Utilized the nystatin-induced open cell-attached patch configuration on A6 cells.
- Compared success rates and stability with excised inside-out patches.
- Performed current-voltage (I-V) relation analysis and determined reversal potential (Erev).
Main Results:
- Open cell-attached patches showed significantly higher success rates (56%) compared to excised patches (9%).
- Both configurations exhibited linear I-V relations and zero Erev in symmetrical Na+ solutions.
- Channel slope conductance and Erev were comparable between the two methods, indicating no nystatin-induced alteration of channel activity.
Conclusions:
- The nystatin-induced open cell-attached patch is a more stable and readily achievable configuration for studying amiloride-sensitive Na+ channels.
- This method provides a viable alternative for experiments involving modulation of cytosolic univalent ions.