This article describes a method for recording afferent nerve activity from the mesenteric afferent nerve in a mouse jejunum segment. The procedure involves distension of the jejunal segment to generate electrical signals for analysis.
Start with a recording chamber containing a dissected segment of a mouse jejunum, a part of the small intestine, submerged in an electrolyte solution and secured with a pin.
The jejunal segment includes surrounding fat tissue with a pre-dissected mesenteric afferent nerve, which transmits signals from the jejunum to the brain.
The segment is connected to the chamber's inflow and outflow ports for solution flow.
Take a suction electrode connected to a micromanipulator and a recording unit.
Fill the suction electrode with the electrolyte solution and position it near the mesenteric afferent nerve.
Apply a gentle suction to pull the nerve with the surrounding fat tissue to form a tight seal.
Close the outflow port while maintaining the inflow to gradually increase the internal pressure and distension of the jejunal segment.
This causes the excitation of the mesenteric afferent nerve around the jejunum, generating an electrical signal.
Record the afferent nerve activity in response to jejunal pressure changes.
In this procedure, using a micromanipulator, lower the tip of the suction electrode into the organ bath. Then, gently aspirate some Krebs solution from the organ bath so that the tip of the electrode is submerged in the Krebs solution. Ensure that the Krebs solution covers the wire electrode inside the suction electrode.
Next, position the tip of the suction electrode immediately next to the transected afferent nerve strand, and draw the transected nerve strand into the capillary over its entire length. Maneuver the tip of the electrode towards the adipose tissue, and aspirate it into the glass capillary with the plunger, thereby mechanically sealing the nerve in the capillary from the contents of the organ bath.
To verify the recording of afferent nerve activity, perform a ramp distension-induced increase in afferent firing by closing the output port to lead to a gradual rise in pressure in the intestinal segment. Only perform the desired experimental protocol when three consecutive ramp distensions yield a reproducible multi-unit discharge.
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