Date of Award

5-2006

Document Type

Thesis

Degree Name

Master of Science (MS)

College/School

College of Science and Mathematics

Department/Program

Biology

Thesis Sponsor/Dissertation Chair/Project Chair

Quinn Vega

Committee Member

James Campanella

Committee Member

Kirsten Monsen

Abstract

RET is a mammalian receptor tyrosine kinase required for proper kidney and neuronal development. RET is expressed as three splice variants named RET9, RET43 and RET51 based on the splice variations at the 3’ end of the gene. All three splice variants of RET are activated by a complex of proteins including a ligand (GFL) and a co-receptor (GFR[alpha]l-4). Although there has been a great deal of work looking at the early stages of RET activation, less is known about the long-term effects, including transcriptional changes that occur in response to RET activation. In this study, transcriptional changes following RET activation were analyzed, paying particular attention to RET, GFR[alpha]2, RET9 and RET51. The expression levels of RET and GFR[alpha]2 were studied using chemilumenescent northern blotting and Quantitative PCR on neuronal cells responsive to GDNF. For analysis of the splice variants, specific primers were designed and RT-PCR was run on RNA isolated from the same cells used previously after the cells were treated with the RET ligand for 0-72 hours. In response to ligand, there was an increase in the levels of RET9 compared to RET51 or to a control gene. This increase was seen at 24 hours and increased up to 72 hours after receptor activation.

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