Sunday, May 16, 2010

Grant Proposal

I’m writing a grant proposal initialed “A Comprehensive Study of Gene Expression in Liver, Intestinal, and Adipose Tissues.” It deals with searching for genes that lead to protein families or enzymes that promote and derogate nutritional absorbencies in the digestive track. This is my PhD thesis. I think this work can be very beneficial to helping fight obesity...I hope it works.
A chicken is an evolutionary species that bridges genetic gaps between birds and mammals, such as higher vertebrates and most importantly humans. Chickens also provide a large amount of human nutrition from meat and eggs throughout the world. Because the chicken is largely used in the human diet, and obesity in humans is on the raise as a worldwide epidemic, there is a need to study the genetic promoters to excessive fat content in chickens. For human health purposes we propose a study on 94 chickens that are divided up into 4 different groups (1.fat males, 2.fat females, 3.lean males, and 4.lean females). All chickens have been feed the same diet. We extract 3 different tissues from these chickens (1.liver, 2.intestinal, and 3.adipose tissue) to compare differences in gene expression (GE) and copy number (CN). The liver secretes bile, which is the fat emulsifier in the digestive track. The small intestine is mainly the largest area of nutrient digestion and produces a mixture of disaccharides, peptides, fatty acids, and monoglycerides. The intestinal tissue in the chicken has not been highly studied in gene expression, even though the final digestion and absorption of food substances occurs mainly in the villi, which line the inner surface of the small intestine. We believe intestinal tissue GE and absorbency is important to study in the over all organisms fat content. Adipose tissue in chickens is a major source of many inflammatory conditions in humans. We will investigate the relationship of abdominal subcutaneous adipose tissues by gene expression to body composition, fat distribution, and metabolic genetic risks that are major contributing factors to human obesity.
How we will do this study:
We will use RNA isolation and RT-PCR. The RNA samples isolated from liver, intestinal and adipose tissue using TRIZOL (Invitrogen) that will be reverse-transcribed to cDNA. A microarray of the entire genome of the chicken will be preformed and areas of special interest will be chosen and primers will be designed for amplification by PCR. Genes that are mainly related to lipid metabolism, fat storage, energy metabolism, and transcription will be of special interest when choosing primer design.