Development of a simple model of extrinsic denervation of the small bowel in mouse.
Small bowel transplantation (SBT) is associated with poorly understood enteric dysfunction. The study of SBT in mice is hindered by the technical difficulty of orthotopic SBT in the mouse. Our aim was to develop an easy preparation of extrinsic denervation of the entire jejunoileum in mice as a model of orthotopic SBT. All neurolymphatic tissues accompanying the superior mesenteric artery (SMA) and vein (SMV) were ligated just distal to the middle colic vessels. The SMA and SMV were then stripped of investing adventitia, and the mesentery to jejunum and colon were transected radially. Jejunum and colon were not transected and reanastomosed. To confirm extrinsic denervation 1, 3, and 6 months later, segments of small bowel were stained for protein gene product 9.5 (PGP9.5) and tyrosine hydroxylase (TH). Tyrosine hydroxylase immunoreactive intensity was then quantified using a semiquantitative analysis. Immunohistochemical fluorescence showed persistence of PGP9.5 immunoreactivity confirming enteric nerves in jejunoileum; however, there was no TH immunoreactivity in jejunoileum in denervated mice despite the expected preservation of TH immunoreactivity in the still-innervated duodenum at 1 month. At 3 months, sparse immunoreactivity for TH was present, and by 6 months, reinnervation of TH-containing nerves appeared similar to controls. Quantification of intensity at each time-point further confirmed this trend. This technique in the mouse accomplishes a complete extrinsic denervation of jejunoileum early postoperatively (1 and 3 months); reinnervation occurs by 6 months. This is an easily learned murine model of orthotopic SBT.
Fatima
Javairiah
J
Department of Surgery, Gastroenterology Research Unit (AL 2-435), Mayo Clinic, 200 First Street SW, Rochester, MN 55905, USA. fatima.javairiah@mayo.edu
Houghton
Scott G
SG
Sarr
Michael G
MG
D000818:Animals; D003714:Denervation / Q000379:methods*; D007082:Ileum / Q000294:innervation / Q000601:surgery*; D007583:Jejunum / Q000294:innervation / Q000601:surgery*; D051379:Mice; D008810:Mice, Inbred C57BL; D023421:Models, Animal