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Enteric neuropathic changes induced by components of the diet and excess of fluoride

Grant number: 13/23862-8
Support Opportunities:Scholarships abroad - Research Internship - Doctorate
Effective date (Start): June 01, 2014
Effective date (End): December 31, 2014
Field of knowledge:Health Sciences - Dentistry
Principal Investigator:Marília Afonso Rabelo Buzalaf
Grantee:Carina Guimarães de Souza Melo
Supervisor: John Barton Furness
Host Institution: Faculdade de Odontologia de Bauru (FOB). Universidade de São Paulo (USP). Bauru , SP, Brazil
Research place: University of Melbourne, Australia  
Associated to the scholarship:11/10233-7 - Evaluation of enteric innervation and proteomic analysis of small intestin of rats exposed to chronic or acute fluoride dose, BP.DR


Nutritional factors can induce profound neuroplastic changes in the Enteric Nervous System (ENS). Dietary components, notably high fats, high cholesterol and advanced glycation end-products (AGEs) are associated with important pathologies. Fluoride (F) in the diet has metabolic effects and it may also affect enteric neurons. Consumption of high-fat diets is implicated in the etiology of obesity, and may lead to gastrointestinal symptoms and physiologic adaptations. Altered innervation of the gastrointestinal tract by the ENS has been shown in various animal models of obesity and there is evidence that a high fat diet causes degenerative changes in enteric neurons. Localization of neuropeptides and proteins in the ENS has been valuable in identifying the classes of neurons that contribute to the control of gastrointestinal function. In this project, we will investigate how diet can lead to enteric neuron damage, and possible dietary improvements that can be applied. In order to accomplish this, segments of stomach, ileum, caecum, and colon collected from C57bl6 mice on a normal chow diet, on a high fat, high cholesterol diet or receiving fluoridated water (50 ppm F - NaF in deionized water) will be evaluated. Special immunofluorescence techniques will be performed using double, and where necessary triple labeling with markers of different myenteric and submucosal neuron types, including NOS, ChAT, VIP, Calretinin, Neurofilament Medium, and Somatostatin. All double labeling will be quantified to determine the relative sizes of different populations and the total neuronal population will be identified with anti-Hu staining. Neuropathic changes will be assessed. Likewise, functional neuro-muscular changes will be recorded using ileal and colon segments mounted in organ baths. Added to this, in order to evaluate the effects of AGEs and RAGEs, tissue collected from all the animal groups will be examined with anti-AGEs and anti-RAGE antibodies. (AU)

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