Perforated Patch-Clamp Recording of Olfactory Sensory Neurons in Mice

Take the severed head of a genetically engineered mouse. The olfactory epithelium in its nose contains olfactory sensory neurons, or OSNs, with fluorophore-tagged odor receptors.

Remove the skin and dissect the head to isolate the septum with attached olfactory epithelium.

Separate and secure the epithelium in a recording chamber.

Using a fluorescence microscope, detect the labeled receptors to target the OSNs' dendritic knob.

Bring a recording pipette with an intracellular solution containing nystatin, a membrane-perforating antibiotic.

Apply positive pressure to avoid pipette clogging. Insert the pipette into the epithelium and approach an OSN.

Switch to negative pressure, forming a seal with the cell membrane. Apply current to maintain the membrane at the resting potential.

Nystatin perforates the membrane, allowing measurement of intracellular ionic currents.

Position a pipette containing an odor stimulant near the OSN. Release the stimulant, which binds to the odor receptors, triggering ion channel opening to allow ion influx. Record the resulting electrophysiological signals.

For this protocol, have mice available between 2 and 4 weeks old. In this presentation, mice expressi