Aug 11, 2019
SPARC - Gastrointestinal myoelectric recordings from the behaving ferret
- Charles C Horn1,
- Derek M. Miller1,
- Stephanie Fulton1,
- Bill J. Yates1,
- Lee E. Fisher1,
- Ameya C. Nanivadekar1
- 1University of Pittsburgh
- SPARCTech. support email: info@neuinfo.org
External link: https://www.biorxiv.org/content/10.1101/607242v2
Protocol Citation: Charles C Horn, Derek M. Miller, Stephanie Fulton, Bill J. Yates, Lee E. Fisher, Ameya C. Nanivadekar 2019. SPARC - Gastrointestinal myoelectric recordings from the behaving ferret. protocols.io https://dx.doi.org/10.17504/protocols.io.6a8hahw
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: August 09, 2019
Last Modified: August 11, 2019
Protocol Integer ID: 26688
Keywords: gastrointestinal tract, stomach, intestine, vagus nerve, electrophysiology, behavior
Abstract
This protocol provides the steps for recording the gastrointestinal myoelectric responses and stimulation of the abdominal vagus nerve from the behaving ferret. These tests occur after an animal is surgically implanted with gastrointestinal and vagus nerve electrodes (see the protocol "SPARC - Chronic implantation of gastrointestinal and vagus nerve electrodes in the ferret" for implantation methods).
Funding: This protocol was developed with funding from the NIH Common Fund’s Stimulating Peripheral Activity to Relieve Conditions (SPARC) program (Award U18TR002205). To learn more about the SPARC program, visit https://sparc.science.
Guidelines
The following guidelines are important:
1) care must be taken to not damage the head connector (e.g., getting debris inside the connection)
2) a ferret will often play with the tether, especially during initial exposure; therefore, the position of the tether will need to be manipulated manually to avoid the animal getting tangled
Materials
MATERIALS
feeding tubeBio-ServCatalog #9012
Emetine dihydrochloride hydrateMerck MilliporeSigma (Sigma-Aldrich)Catalog #45160
Room setup
Room setup
Cover floor of behavioral test chamber with layer of Alpha-dri bedding. The floor of the plexiglas chamber is 51 x 51 cm (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3348962/)
Turn off all lights except lamp over testing chamber.
Set up video camera directly above chamber so that entire area is visible with minimal shadows. Set up a second camera to view the feeding tube from the side for feeding trials to track volume consumed.
Equipment
video camera
NAME
camera
TYPE
Sony
BRAND
HDR-XR550V
SKU
Suspend tether ~50 cm above chamber and secure with rubber bands to provide elasticity as the animal moves.
Keep the room quiet during recording except for white noise.
Animal
Animal
Animal should be fasted 3 hours before starting recording.
Place animal in chamber and allow 10 minutes for acclimation.
Connect animal to tether.
During testing, hold the tether straight above the animal out of its reach, but with some slack. If the animal tries to grab the tether, especially during adaptation, a plastic straw can be inserted through the breathing holes in the side of the chamber to distract the animal from the tether.
Recording: Baseline control
Recording: Baseline control
Record electrode impedances before the recording begins.
Equipment
Grapevine Neural Interface Processor and a Nano2 recording headstage
NAME
electrophysiological recording and stimulation
TYPE
Ripple LLC
BRAND
Grapevine Neural Interface Processor and a Nano2 r
SKU
Electrophysiological recording and video recording should be started at the same time.
Equipment
Media Recorder
NAME
video to computer recording system
TYPE
Noldus Information Technology
BRAND
MediaRecorder
SKU
LINK
Record 1 h of baseline data for control days.
Recording: Feeding
Recording: Feeding
For feeding trials, introduce Ensure liquid diet in a graduated tube (100 ml Richter glass feeding tube) attached to the side of the chamber at 10 min after the start of recording. Remove the feeding tube 45 min after presentation. Finish with 10 min of baseline recording.
Equipment
Grapevine Neural Interface Processor and a Nano2 recording headstage
NAME
electrophysiological recording and stimulation
TYPE
Ripple LLC
BRAND
Grapevine Neural Interface Processor and a Nano2 r
SKU
Recording: Feeding with vagus stimulation
Recording: Feeding with vagus stimulation
For feeding with stimulation trials, introduce Ensure at the same time as in Step 13. Start stimulation 2 minutes after food introduction. Stop stimulation and remove food at the same time, i.e., minute 45 in the recording. Finish with 10 minutes of baseline
Equipment
Grapevine Neural Interface Processor and a Nano2 recording headstage
NAME
electrophysiological recording and stimulation
TYPE
Ripple LLC
BRAND
Grapevine Neural Interface Processor and a Nano2 r
SKU
Recording: Emetiine (emesis testing)
Recording: Emetiine (emesis testing)
For emetine trials, begin infusion at the 10 min in the recording. Infusion rate is 10ml/min. Record for 1 hour after infusion has ended.
Equipment
Grapevine Neural Interface Processor and a Nano2 recording headstage
NAME
electrophysiological recording and stimulation
TYPE
Ripple LLC
BRAND
Grapevine Neural Interface Processor and a Nano2 r
SKU
After recording is done, animal should be monitored until there is no further emesis.
Recording: Water control
Recording: Water control
For water control trials, follow Step 15.
Equipment
Grapevine Neural Interface Processor and a Nano2 recording headstage
NAME
electrophysiological recording and stimulation
TYPE
Ripple LLC
BRAND
Grapevine Neural Interface Processor and a Nano2 r
SKU
End
End
Stop electrophysiological and video recordings at the same time.
Measure impedances at the end of each recording.
Equipment
Grapevine Neural Interface Processor and a Nano2 recording headstage
NAME
electrophysiological recording and stimulation
TYPE
Ripple LLC
BRAND
Grapevine Neural Interface Processor and a Nano2 r
SKU