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Whole-cell recording from non-dissociated taste cells in mouse taste bud
T Miyamoto1, T Miyazaki, Y Okada
1Department of Physiology, Nagasaki University School of Dentistry, Japan.
Insights
This study details a new method for whole-cell recording in mouse taste cells. This technique allows for simultaneous patch-clamping, cell visualization, and localized taste stimulation.
Area of Science:
- Neuroscience
- Sensory Biology
- Physiology
Background:
- Understanding taste perception requires studying individual taste cells.
- Current methods often involve dissociating cells, which can alter their native properties.
Purpose of the Study:
- To develop a novel method for whole-cell recording from intact, non-dissociated taste cells within mouse taste buds.
- To enable simultaneous electrophysiological recording, morphological visualization, and localized stimulation of taste cells.
Main Methods:
- A technique involving enzymatic digestion (elastase) and specific Tyrode solutions to isolate taste buds with minimal tissue disruption.
- Utilizing holding pipettes for orientation and patch pipettes with Lucifer Yellow for whole-cell recording.
- Employing a third pipette for localized taste stimuli or blocker application near the taste pore under continuous solution flow.
Main Results:
- Successfully established whole-cell configuration in non-dissociated taste cells.
- Achieved simultaneous patch-clamping, visualization of taste cell morphology, and localized taste stimulation.
- Obtained rapid electrophysiological responses to salt stimuli, demonstrating the viability of the preparation.
Conclusions:
- The described method provides a robust approach for studying taste cell function in their native environment.
- This technique preserves the microenvironment of the taste organ, crucial for accurate physiological measurements.
- Facilitates detailed investigation into taste cell responses and mechanisms.
Abstract:
A method for the whole-cell recording from non-dissociated taste cells within mouse taste bud is described. The lingual epithelial sheet containing the taste buds was peeled free from the tongue by injecting a proteolytic enzyme, elastase, under the lingual epithelium and by incubating it in normal Tyrode solution at 30 degrees C. The preparation consisting of a taste bud and a small piece of the lingual epithelium was obtained by further the incubation in divalent cation-free Tyrode solution. After holding the small piece of the epithelium by a holding pipette loaded with continuous negative pressure for keeping the orientation of the taste bud, whole-cell configuration was established in a non-dissociated taste cell within the taste bud with a patch pipette containing Lucifer Yellow. Taste stimuli or blockers were applied from the third pipette placed near the taste pore under the continuous flow of bathing solution. Under this condition, we could simultaneously accomplish patch-clamping, visualization of taste cell morphology, localized taste stimulation and maintenance of microenvironment around the taste organ. Rapid responses to a relatively high concentration of salt stimuli were also obtained.
