Project description:Analysis of gene expression profiles of epididymal fat from DIO rats We applied a comparative functional genomics approach to evaluate diet-induced obese (DIO) rats as an obesity model Keywords: single time point, comparison control animal vs. diet induced obese animal
Project description:According to different feeding and treatment conditions, 36 C57BL/6JC rats were randomly divided into normal diet group (WC group), high fat diet group (WF group) and high fat diet + silibinin group (WS group). TMT combined with LC-MS/MS were used to study the expression of WAT in epididymis of HFD-induced obese rats and normal diet rats. Gene Ontology, InterPro and KEGG databases were used to analyze the cellular processes, the biological processes, the corresponding molecular functions and the network molecular mechanisms involved
Project description:Background: Post-menopausal obesity is an established risk factor for breast cancer. Consumption of diets high in fat is known to be highly correlated with obesity. In this, we sought to evaluate the interaction(s) between high fat diet, weight gain and mammary carcinogenesis using an obese-resistant and obese-prone rat model with direct correlates to human disease. Methods: Female obese-prone (OP) and obese-resistant (OR) weanling rats were placed on either a low fat (10% kcal) or a high fat (39% kcal) n-6 polyunsaturated (PUFA) safflower diet for 30 days. At post natal day (PND) 50, global gene expression profiling was performed on microdissected mammary epithlelium from one cohort of rats and another cohort of rats were given a single oral gavage of either 7,12-dimethylbenz[a]anthracene (DMBA at 14 mg/kg) or vehicle. Rats were then maintained on the diets and body weights, food consumption and development of mammary lesions were monitored weekly. Results: The DMBA-treated OR rats on the 39% safflower diet had significantly greater incidence of ductal carcinoma-in-situ (DCIS) lesions and significantly greater DCIS multiplicity than DMBA-treated OR rats on the 10% safflower diet. These differences were not seen in the OP strain. Gene expression analysis of mammary ductal epithelium from OR rats on the high fat diet showed significant upregulation of proliferation-related genes compared to those consuming the low fat safflower diet. Again, these differences were not seen in the OP strain. Conclusion: Our findings indicate that consumption of high fat safflower diet enhances mammary carcinogenesis in an OR rat strain through increased proliferation of mammary epithelium at the time of exposure, but not in the OP rat strain. Thus, the diet-induced increase in sensitivity was strain-specific and independent of weight gain or obesity level. Female obese-prone (OP) and obese-resistant (OR) weanling rats were placed on either a low fat (10% kcal) or a high fat (39% kcal) n-6 polyunsaturated (PUFA) safflower diet for 30 days. At post natal day (PND) 50, global gene expression profiling was performed on microdissected mammary epithlelium from one cohort of rats and another cohort of rats were given a single oral gavage of either 7,12-dimethylbenz[a]anthracene (DMBA at 14 mg/kg) or vehicle. Rats were then maintained on the diets and body weights, food consumption and development of mammary lesions were monitored weekly.
Project description:Analysis of gene expression profiles of epididymal fat from DIO rats; We applied a comparative functional genomics approach to evaluate diet-induced obese (DIO) rats as an obesity model Experiment Overall Design: Gene expression profiles from DIO and lean rats were generated and compared. A private colony of male Long-Evans rats was set up at Harlan (Harlan-Sprague Dawley, Indianapolis, IN). The animals had been on either a high fat/high sucrose diet (TD95217, Harlan Takled, Madison, WI) or a regular chow (Purina 5008; Ralston-Purina, St. Louis, MO) since weaning. Rats were shipped at 14-weeks of age to our facility and maintained on a 12:12-hour light-dark photoperiod (light on 10:00 PM). They were individually housed throughout the study at ambient temperature, and had free access to diets and water. After 2 weeks acclimation to the facility, body weight of the rats and weight of food consumed in the last 7 days were measured once a week. Body fat mass was analyzed by nuclear magnetic resonance (NMR) using an Echo Medical System (Houston, TX) instrument 3 times throughout the study. Fat free mass is the difference between body mass and fat mass. All animal studies were conducted in compliance with approved institutional animal care and use protocols according to NIH guidelines (NIH publication No. 86-23, 1985). OGTT was conducted after an overnight fasting when animals were 21 weeks of age. After a baseline blood collection (time 0), animals were dosed orally with 1.5 gram/kg glucose. Subsequence blood samples were collected at 30, 60, and 120 minutes post glucose challenge for the determination of plasma glucose and insulin levels. At 25 weeks of age, blood was collected by tail bleeding after overnight fasting. Rats were killed a week later at 09:00am by rapid decapitation. Trunk blood was collected and epididymal fat tissue excised. Various circulating metabolic parameters were measured under fasted (week 25) and fed conditions (week 26). Total RNA from rat epididymal fat was isolated with RNA STAT-60 (Tel-Test) according to the manufacturer's protocol. 5 micrograms of total RNA were labeled and hybridized to Affymetrix RAE230_2 arrays according to the Affymetrix protocol.
Project description:Purpose: Successive paternal generations of unhealthy diet induce a fat mass increase. The goal of this study is to compare epididymal adipose tissue transcriptome profiling (RNA-seq) of mice fed either a control-Diet or a High-Fat-Diet for One or Five successive generations. The aim of the study is to identify differentially expressed RNA in epididymal adipose tissue of obese males which might be involved in the exacerbation of induced by the maintenance of High Fat Diet feeding for 5 successive generations. Methods: Epididymal adipose tissue mRNA profiles of 18-months-old mice fed either a control-Diet or a Western-Diet for One or Five successive generations were generated by deep sequencing, in triplicate, using Illumina GAIIx. The sequence reads that passed quality filters were quantitavely analyzed. Reads abundance was evaluated for each gene followed by annotation versus mouse GTF by using the featureCounts function.The R package Edger was used in order to normalize the reads and to identify differentially expressed (DE) genes. Results: RNA-seq data revealed a specific enrichment in gene related to metabolic syndrome traits in male mice obtained after successive paternal generations of Western-Diet.