North Carolina State University Undergraduate Symposium





2012- 21st Annual NC State Undergraduate Research Symposium

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Session Time : 4/10/12 12:15 PM - 4/10/12 1:30 PM
Content Area : Animal Science
Poster Appointment: NONE
Student Presenters :       
Taylor Alexander Treadaway Genetics
Mentors and/or Co-Authors :
Melissa Ashwell Animal Science
Abstract Title : Gene Level Pain Expression in Feline Associated Degenerative Joint Disease
Abstract :
Degenerative joint disease is a crippling disorder that results in painful movement, and stiffness in the joints.  This crippling condition is not very well understood in cats, which makes their treatment difficult. Therefore we sought to determine if feline sufferers of degenerative joint disease express pain at the genetic level in a significantly different manner from disease free cats by examining expression of genes thought to be associated with pain. My contribution to this project was to identify the most stable reference genes in both cats with and without the disease. The best reference genes are expressed at stable levels across all samples to give a stable platform for comparison. We gathered dorsal root ganglia from cats determined to have degenerative joint disease, and ganglia from control animals. From those samples we isolated total RNA that was converted to cDNA. Using the cDNA from both affected and unaffected cats, we ran real time PCR temperature gradients with primer pairs to determine their optimal annealing temperatures. We then examined expression of each gene in all the cDNA samples to determine which genes are the most stably expressed. To identify the most stably expressed reference genes, we used software programs designed to determine the best reference genes. Based on results from these two programs, we determined the three most stably expressed genes across all samples are RPL30, HMBS, and GUSB. With these results we can further analyze our feline samples to determine if genes found to encode for pain in prior rodent studies have significantly different gene expression levels in diseased versus healthy samples in the cat.