BACKGROUND: Our aim was to develop and validate a technically easy, reliable, and reproducible method of complete jejunoileal denervation in the rat to allow study of the physiologic effects of intestinal transplantation devoid of immunologic phenomena and ischemia/reperfusion injury. MATERIALS AND METHODS: Six adult Sprague-Dawley rats underwent transection and reanastomosis of the proximal jejunum and proximal colon, transection of all neurolymphatic tissues at the base of the mesentery, stripping adventitia off the superior mesenteric artery and vein, and radial transection of the intervening mesenteries, thereby denervating the jejunoileum in situ without disrupting blood flow. Three rats each were sacrificed 1 and 6 months later. Intestinal smooth muscle from the still-innervated duodenum and the denervated jejunum, mid-small bowel, and ileum was compared to corresponding tissues from a normal rat for tyrosine hydroxylase immunohistochemistry, a marker of extrinsic innervation. RESULTS: One and six months after denervation, all duodenal samples demonstrated normal tyrosine hydroxylase immunostaining. In contrast, tyrosine hydroxylase immunoreactivity was undetectable in jejunum, mid-small bowel, or ileum of rats at 1 month and 2 of the 3 rats at 6 months; 1 rat at 6 months had low levels of tyrosine hydroxylase immunoreactivity at the mesenteric border of jejunum and mid small bowel. CONCLUSION: This simple technique of in situ neural isolation effectively and reproducibly achieves complete extrinsic denervation of the entire rat jejunoileum. Low levels of neural regeneration may be present 6 months after denervation.
BACKGROUND: Our aim was to develop and validate a technically easy, reliable, and reproducible method of complete jejunoileal denervation in the rat to allow study of the physiologic effects of intestinal transplantation devoid of immunologic phenomena and ischemia/reperfusion injury. MATERIALS AND METHODS: Six adult Sprague-Dawley rats underwent transection and reanastomosis of the proximal jejunum and proximal colon, transection of all neurolymphatic tissues at the base of the mesentery, stripping adventitia off the superior mesenteric artery and vein, and radial transection of the intervening mesenteries, thereby denervating the jejunoileum in situ without disrupting blood flow. Three rats each were sacrificed 1 and 6 months later. Intestinal smooth muscle from the still-innervated duodenum and the denervated jejunum, mid-small bowel, and ileum was compared to corresponding tissues from a normal rat for tyrosine hydroxylase immunohistochemistry, a marker of extrinsic innervation. RESULTS: One and six months after denervation, all duodenal samples demonstrated normal tyrosine hydroxylase immunostaining. In contrast, tyrosine hydroxylase immunoreactivity was undetectable in jejunum, mid-small bowel, or ileum of rats at 1 month and 2 of the 3 rats at 6 months; 1 rat at 6 months had low levels of tyrosine hydroxylase immunoreactivity at the mesenteric border of jejunum and mid small bowel. CONCLUSION: This simple technique of in situ neural isolation effectively and reproducibly achieves complete extrinsic denervation of the entire rat jejunoileum. Low levels of neural regeneration may be present 6 months after denervation.