S&T-31 TRANSCRIPTIONAL REGULATION OF CORTICOTROPIN RELEASING FACTOR

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You have accessJournal of UrologyScience & Technology Posters1 Apr 2016S&T-31 TRANSCRIPTIONAL REGULATION OF CORTICOTROPIN RELEASING FACTOR Lizath Aguiniga, Anthony Schaeffer, and David Klumpp Lizath AguinigaLizath Aguiniga More articles by this author , Anthony SchaefferAnthony Schaeffer More articles by this author , and David KlumppDavid Klumpp More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2016.02.2860AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Interstitial cystitis (IC) patients suffer from chronic pelvic pain and bladder dysfunction. IC patients have altered cortisol levels suggesting dysregulation of the hypothalamic-pituitary adrenal (HPA) axis and suffer from exacerbated symptoms in response to high stress. Corticotropin-releasing factor (CRF) is the initiator of the HPA axis and mediates stress responses and voiding control, where increased CRF levels in Barrington′s nucleus induce bladder dysfunction. Arachidonic acid (AA) metabolites have been shown to induce CRF expression, however the transcriptional mediators of this modulation are unknown. We are investigating which transcription factors mediate AA-induced CRF gene expression. METHODS We used MIRAGE software to identify candidate transcription factor binding sites in a 1kb region of the human CRF gene promoter. We identified a peroxisome proliferator activated hormone response element (PPRE) and two Xenobiotic Responsive Element (XRE) sites as candidate mediators of AA-dependent CRF induction. Site directed mutations of the PPRE and XRE sites were generated in a CRF luciferase reporter plasmid. We evaluated the WT CRF promoter and the mutants for their responses to AA. We also over-expressed transcription factors AhR and PPAR gamma and evaluated AA-induced CRF expression in HEK 293T cells. We used the hypothalamic cell line N42, which has functional CRF expression to assess the role of AhR. We over-expressed AhR, induced with AA and used RT PCR to quantify CRF expression. RESULTS AA induction in the PPRE mutant resulted in increased CRF promoter activity compared to WT, whereas XRE1 had decreased activity. The mutation of both XRE1 and XRE2 resulted in decreased responsiveness to AA. Over expression of PPAR gamma alone showed no change, while over expression of AhR alone showed a significant increase in AA-induced CRF expression; this was inhibited by over-expression of both AhR and PPAR gamma in HEK 293T cells. Over expression of AhR in N42 cells resulted in increased CRF mRNA in response to AA induction. CONCLUSIONS These results suggest AhR binding to the XRE sites modulates AA-dependent CRF gene expression. PPAR gamma inhibits AA-dependent CRF gene expression. Continued studies will use mouse models to examine the in vivo role of AhR and PPAR gamma in modulating voiding in response to stress. © 2016FiguresReferencesRelatedDetails Volume 195Issue 4SApril 2016Page: e321 Advertisement Copyright & Permissions© 2016MetricsAuthor Information Lizath Aguiniga More articles by this author Anthony Schaeffer More articles by this author David Klumpp More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...