Jan 15, 2024
Protocol for vagus nerve stimulation in awake pigs (subject-3)
- Oliver Armitage1
- 1BIOS Health
Protocol Citation: Oliver Armitage 2024. Protocol for vagus nerve stimulation in awake pigs (subject-3). protocols.io https://dx.doi.org/10.17504/protocols.io.rm7vzb692vx1/v1
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: January 03, 2023
Last Modified: January 15, 2024
Protocol Integer ID: 74702
Funders Acknowledgement:
SPARC
Grant ID: OTA-20-004
Abstract
The Vagus nerve innervates a number of thoracic and visceral organs. Exogenous nervous signal, for example, Vagus nerve stimulation (VNS) provides a route to modulating their function for therapeutic purposes. However, the relationship between VNS parameters and organ responses is poorly understood as evidenced by a number of failed human trials, for example, in the heart failure space.
This protocol is for a chronic study that builds upon a previous acute study (https://doi.org/10.26275/dm5l-tspb) in which the pigs were under anesthesia. It maps VNS parameters to their immediate neural and cardiac effects in 2 freely moving pigs. By exploring the changes in heart rate responses from acute to chronic periods, we aim to gain a more comprehensive understanding of the effects of VNS on organ function.
Subjects
Subjects
Female swine of adult age and weight between 35—55 kg for the experimental design were used.
Surgery
Surgery
Measure the weight of the pig, then administer anesthesia. Intubate the animal and start mechanical ventilation.
Position pig prone and mark neck where the skin implant is to be placed along with the incisions on the neck.
Cut out circle for skin implant to go through and make an incision where the implant will be inserted.
Place skin implant in position and sew into muscle and skin.
Tunnel the cuffs and leads to an open incision halfway down the neck.
Position the pig supine with neck extended and make a single incision through the skin and superficial fat layers between the mandible and sternal notch. Use blunt dissection to expose the carotid sheath and isolate the vagus nerve from the carotid artery.
Use blunt dissection to locate the superior and recurrent laryngeal branches of the vagus nerve.
Use blunt dissection to expose as much length of the vagus trunk as possible. Ideally at least 8 cm from the superior laryngeal branch insertion point to a more caudal location.
Tunnel the leads and cuffs from the incision in the neck to where the vagus nerve is exposed.
Place cuffs on vagus nerve such that there is at least 4 cm between each cuff, but ideally 5.5 cm.
Sew EMG patch on laryngeal muscle and ground lead in a pocket next to the caudal cuff. Coil the ECG lead and place in a pocket on the right pectoral. Tack down the wires.
Validate system works before closing incisions.
Close incisions.
Stimulation Protocol
Stimulation Protocol
Connect BIOS Neurotool software to implant.
Set the the following stimulation parameters in BIOS Neurotool software:
- Pulse width: 260us
- Train duration: 3s
- Stimulation shape: monophasic
- Polarity: cathodic
Conduct a grid search by doing the cartesian product of the frequency and current parameters.
- Frequency: {2Hz, 5Hz, 10Hz, 20Hz, 30Hz}
- Current: {0.05mA, 0.5mA, 1mA, 1.5mA, 2.5mA}
After each stimulation, wait for the physiological signals to return to baseline.