Mechanosensitive ion channels investigated simultaneously by scanning probe microscopy and patch clamp

Area of Science:

• Biophysics
• Cell Biology
• Neuroscience

Background:

• Mechanosensitive ion channels are crucial for sensing mechanical stimuli like touch, balance, and sound.
• Understanding these channels in sensory systems, such as auditory hair cells, is vital for explaining mechanotransduction.

Purpose of the Study:

• To present a new experimental strategy for well-defined access to single mechanosensitive ion channels in living cells.
• To detail the investigation of mechanosensitive transduction channels in cochlear hair cells using a combination of advanced techniques.

Main Methods:

• Integration of atomic force microscopy (AFM) for localized mechanical stimulation.
• Utilization of patch clamp technique for recording ion channel current responses.
• Employing differential interference contrast (DIC) microscopy with specialized optics for high-resolution imaging.

Main Results:

• Demonstration of a feasible experimental setup combining AFM, patch clamp, and DIC microscopy.
• Detailed protocols for preparing and investigating the organ of Corti using AFM.
• Successful adaptation of AFM mechanical and electrical designs for integration with light microscopy.

Conclusions:

• The presented strategy provides unprecedented access to single mechanosensitive ion channels in their native cellular environment.
• This methodology is particularly valuable for studying auditory mechanotransduction in cochlear hair cells.
• The technical advancements facilitate future research into the function and modulation of mechanosensitive channels.