Validation and Localization of Restricted Gene Expression in the Developing Prostate

Date
2009-05
Authors
Frank, Sander Barkley
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University of Delaware
Abstract
The prostate gland is a significant source of male genitourinary tract morbidity. As developmental processes share several features in common with metastatic cancer, I hypothesize that a better understanding of genes involved in prostate morphogenesis may identify possible targets for therapeutic intervention. The prostate is derived from the urogenital sinus (UGS), which can be separated into four subdomains based on cellular compartment and dorsal-ventral patterning: epithelium/mesenchyme (UGE/UGM) and dorsal/ventral halves (UGD/UGV). Each region develops into specific lobes of the prostate with unique structural and biochemical properties. My research sought to validate differential gene expression associated with each of these UGS subcompartments as determined by a survey of more than 15,000 cDNA fragments in a custom microarray covering a timed course of prostate development. Results identified 530 genes (3.5%) in UGE/UGM, and 35 (0.23%) in UGD/UGV that exhibit spatially restricted expression in UGS subdomains. To validate the microarray results, male UGS were separated either into UGE/UGM or UGD/UGV subcompartments. Total RNA was extracted and used to synthesize cDNA templates for each subdomain. Quantitative real time reverse transcription polymerase chain reaction (Q-PCR) was performed to observe relative mRNA levels and confirm localization for 10/12 primary and 6/10 secondary compartmental genes. Immunofluorescence (IF) was used to localize 5 genes in the predicted subdomains. With this research I have established a map of regional gene expression in the murine UGS and identified candidate genes for further study and analysis.
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