TITLE: Implantation of a pelvic nerve array in rats
AUTHORS: James Fallon, Sophie Payne, Janet Keast, Peregrine Osborne

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">A pelvic nerve array is custom designed for implantation onto the pelvic nerve of male rats. This device is used for long-term (recovery) experiments. The device can electrically stimulate the pelvic nerve as well as record electrically evoked or spontaneous neural activity. Previously, we have successfully implanted rats for 2 months with no reports of biological adverse complications or damage sustained to the array. The surgical procedure is performed under anesthesia and should incorporate all local requirements for standards of animal experimentation, including methods of anesthesia, surgical environment, and post-operative monitoring and care. </div></div>

[STEPS]
?. [Surgical Preparation]
Prior to surgery, pelvic nerve arrays are sterilised through a sonication process and autoclaving.
?. [Surgical Preparation]
Sonication cycles: 15 mins in 5% Pyroneg, 5 mins distilled water, 5 mins in distilled water, 10 mins in 96% ethanol, 5 mins in distilled water, 5 mins in distilled water
?. [Surgical Preparation]
Autoclave
?. [Surgical Preparation]
For surgery, anesthetise (2.5% isoflurane in oxygen, or as required for maintenance) the animal (adult male rat Sprague-Dawley, 8-10 weeks)
?. [Surgical Preparation]
Apply eye lubricant and place animal on a heat pad
?. [Surgical Preparation]
Shave and clean (betadine) the ventral abdomen and the dorsal-lumbar back
?. [Surgery]
Make an incision to the dorsal-lumbar aspect of the back (this will be the exit site for the percutaneous electrode connector).
?. [Surgery]
Make a ventral midline incision in the skin overlying the region of the bladder
?. [Surgery]
Using hemostats, subcutaneously tunnel the array from the dorsal incision site through to the ventral incision site.
?. [Surgery]
Make an incision in the ventral midline of the abdominal cavity to expose the bladder.
?. [Surgery]
Make a second, small (1 cm) incision (lateral to the midline incision) to the abdominal muscle on the same side as lead wire, and feed the array through this secondary incision.
?. [Surgery]
Locate the left or the right prostate and gently retract the lobe to reveal the pelvic ganglia and connecting nerves (including the pelvic nerve)
?. [Surgery]
Clearly identify the pelvic nerve
?. [Surgery]
The array is placed over the top of the pelvic nerve and secured in place to the connective tissue of the prostate using sutures (7’0 silk).
?. [Surgery]
Exteriorise the lead wire through the abdominal cavity (via the second small incision).
?. [Surgery]
At this stage of the surgery, a bladder catheter can be implanted according to the protocol described in 'Cystometry for Awake Rats'
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?. [Surgery]
Close the abdominal cavity and skin incision sites with sutures.
?. [Surgery]
Suture (2’0 silk) the percutaneous pedestal silicone-Dacron embedded mesh to the underlying muscle and connective tissue of the dorsal midline near the neck.
?. [Surgery]
Close the skin incision site with sutures.
?. [Surgery]
Administer analgesic and sterile Hartmann’s solution.
?. [Surgery]
During post-surgery management, the skin around the percutaneous plug is cleaned with betadine every 2 weeks to ensure an infection does not develop. Rats can be awake, but gently restrained, during this maintenance procedure.