The following protocol was submitted on behalf of the authors from the Bornstein lab by the SPARC... more The following protocol was submitted on behalf of the authors from the Bornstein lab by the SPARC project. This protocol describes methods for standard intracellular recording from mouse myenteric neurons impaled with intracellular electrodes containing biocytin, followed by processing for immunohistochemistry to map the projections of the neurites of enteric neurons. Methods described in this protocol are adapted from decades of studies on guinea-pig enteric neurons and more recent analyses of mouse duodenal myenteric neurons and colonic submucosal neurons. The immunofluorescence is used to reveal either calretinin or neuronal nitric oxide synthase (nNOS), as well as the enteric neurons in the proximal colon of the mouse. The methods can be generalized to whole mount preparations from any gut region in any species. Mice were sacrificed by cervical dislocation, a procedure approved by the University Melbourne Animal Experimentation Ethics committee. The experimental procedures shoul...
Background The autonomic nervous system (ANS) is thought to play a critical role in the anti-infl... more Background The autonomic nervous system (ANS) is thought to play a critical role in the anti-inflammatory reflex pathway in acute colitis via its interaction with the spleen and colon. Inflammation in the intestine is associated with a blunting of vagal signaling and increased sympathetic activity. As a corollary, methods to restore sympatho-vagal balance are being investigated as therapeutic strategies for the treatment of intestinal inflammation. Nevertheless, it is indefinite whether these autonomic signaling adaptations in colitis are detrimental or beneficial to controlling intestinal inflammation. In this study, models of moderate and severe chronic colitis are utilized to resolve the correlations between sympatho-vagal signaling and the severity of intestinal inflammation. Methods Spleens and colons were collected from Winnie (moderate colitis), Winnie-Prolapse (severe colitis), and control C57BL/6 mice. Changes to the size and histomorphology of spleens were evaluated. Flow ...
The following protocol was submitted on behalf of the authors from the Bornstein lab by the SPARC... more The following protocol was submitted on behalf of the authors from the Bornstein lab by the SPARC project. This protocol describes methods for standard intracellular recording from mouse myenteric neurons impaled with intracellular electrodes containing biocytin, followed by processing for immunohistochemistry to map the projections of the neurites of enteric neurons. Methods described in this protocol are adapted from decades of studies on guinea-pig enteric neurons and more recent analyses of mouse duodenal myenteric neurons and colonic submucosal neurons. The immunofluorescence is used to reveal either calretinin or neuronal nitric oxide synthase (nNOS), as well as the enteric neurons in the proximal colon of the mouse. The methods can be generalized to whole mount preparations from any gut region in any species. Mice were sacrificed by cervical dislocation, a procedure approved by the University Melbourne Animal Experimentation Ethics committee. The experimental procedures shoul...
Background The autonomic nervous system (ANS) is thought to play a critical role in the anti-infl... more Background The autonomic nervous system (ANS) is thought to play a critical role in the anti-inflammatory reflex pathway in acute colitis via its interaction with the spleen and colon. Inflammation in the intestine is associated with a blunting of vagal signaling and increased sympathetic activity. As a corollary, methods to restore sympatho-vagal balance are being investigated as therapeutic strategies for the treatment of intestinal inflammation. Nevertheless, it is indefinite whether these autonomic signaling adaptations in colitis are detrimental or beneficial to controlling intestinal inflammation. In this study, models of moderate and severe chronic colitis are utilized to resolve the correlations between sympatho-vagal signaling and the severity of intestinal inflammation. Methods Spleens and colons were collected from Winnie (moderate colitis), Winnie-Prolapse (severe colitis), and control C57BL/6 mice. Changes to the size and histomorphology of spleens were evaluated. Flow ...
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Papers by Joel Bornstein