Methylation and expression patterns of the Pomc gene in the offspring of rats subjected to deficient and supplemented with folic acid diets

Author(s)

Bruna Morais Faleiros de Paula

• Document type: Master's Dissertation
• Press: Ribeirão Preto.
• Institution: Universidade de São Paulo (USP). Faculdade de Medicina de Ribeirão Preto (PCARP/BC)
• Defense date: 2016-11-22

Examining board members

Helio Vannucchi; Carla Barbosa Nonino; Sérgio Alberto Rupp de Paiva

• Advisor: Helio Vannucchi

Abstract

Epigenetic mechanisms are essential for proper development in mammals, and that changes at this stage may lead to various metabolic disorders such as obesity. Currently obesity is a serious problem of public health worldwide, has a multifactorial origin involving both environmental and genetic factors. There are some genes that are involved with obesity, such as the proopiomelanocortin gene (POMC). The aim of this research project is to investigate the expression and methylation patterns of the Pomc gene in the offspring of rats subjected to deficient and supplemented diets with folic acid. Animals used were Wistar rats. The study involved males and females pups (n = 24) that were weaned at the same diet their mothers, three treatment groups, control group (2,0 mg/kg of folic acid), deficient group (0,5 mg/kg of folic acid) and the supplemented group (8,0 mg/kg of folic acid). The arcuate nucleus of the hypothalamus tissue were collected, from which was extracted DNA, RNA and proteins using commercially available kits following the manufacturer\'s protocol. The DNA methylation pattern analysis was performed. The mRNA was used for the analysis of gene expression by real time PCR, the TaqMan (Life Technologies (TM)) system. The proteomic study was carried out by Western blotting. In general, we found that the body weight of the male offspring showed no statistical difference between the groups. The feed intake of folic acid deficient group was statistically (p = 0.03) higher than the control group. In relation to female offspring was observed that the body weight of the supplemented group was statistically (p = 0.01) higher than the control group, and related to feed intake, there was no statistically significant difference between the treatment groups. The analysis of brain weight, gene expression, methylation and protein expression of Pomc no significant statistical differences among treatment groups of both sexes. Concluded that the intervention diets with different folic acid concentrations did not cause significant changes in the offspring compared to the study of proteomics and methylation and expression patterns of the Pomc gene. As for the body weight and feed consumption of animals it showed that supplementation with folic acid during pregnancy and post weaning was able to alter these two parameters with differing response between males and females in the adult offspring. (AU)

• FAPESP's process: 14/15357-4 - Methylation and expression patterns of the POMC gene in the offspring of rats subjected to diets deficient and supplemented with folic acid
• Grantee: Bruna Morais Faleiros de Paula
• Support Opportunities: Scholarships in Brazil - Master