Abstract
Alterations in brain serotonin levels are implicated in major depression and are regulated by tryptophan hydroxylase-2 (TPH2). To study its regulation, we measured TPH2 RNA by quantitative RT-PCR in differentiated serotonergic rat raphe RN46A and GH4C1 pituitary cells, which express TPH2. Upon calcium mobilization using KCl (40 mmol/L), TPH2 RNA was rapidly (1 h) and strongly (> 10-fold) induced in differentiated RN46A cells, but not in GH4C1 cells. This effect was blocked by actinomycin D, implicating transcriptional activation. Similarly, calcium ionophore ionomycin induced TPH2 RNA by threefold in RN46A cells. To address the promoter sites involved, the transcription start site was identified and a series of TATA-containing TPH2 promoter-luciferase constructs were analyzed. In differentiated RN46A cells, the TPH2 promoter was induced 2.5-fold by ionomycin, similar to its action on TPH2 RNA. By contrast, ionomycin had no effect on TPH2 promoter activity in GH4C1 cells or TPH2-negative L6 myoblasts. Ionomycin sensitivity was localized to within 88 bp of the start site, containing putative CCATT-enhancer binding protein element, activator protein-1 and -2 (AP-1, AP-2) elements. These results are the first to identify calcium-mediated regulation of the proximal TPH2 promoter as critical for cell-specific TPH2 expression.
Published Version (
Free)
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
