title: Estradiol induces transcriptional and posttranscriptional modifications in versican expression in the mouse uterus.
abstract: We have previously shown the differential expression of versican in the mouse uterus under ovarian hormone influence. We also demonstrated there is not a direct correlation between mRNA levels and protein expression, suggesting posttranscriptional events, such as alteration in mRNA stability. This posttranscriptional effect may result in the elongation and stabilization of transcripts poly(A) tail. Thus, the aim of this study was to analyze whether estradiol (E2) regulates versican mRNA stability and expression in a dose-related and time-dependent manner. For this purpose female mice were ovariectomized and treated with a single injection of 0.1 or 10 μg E2. To block transcription a group of females received a single injection of alpha-amanitin before hormone administration. Uterine tissues were collected 30 min, 1, 3, 6, 12 and 24 h after treatments and processed for quantitative real time PCR (qPCR), RACE-PAT Assay and immunohistochemistry. qPCR showed that versican mRNA levels are higher than control from 3 to 24 h after E2 administration, whereas after transcription inhibition versican mRNA unexpectedly increases within 3 h, which can be explained when transcriptional blockers alter the degradation rate of the transcript, resulting in the superinduction of this mRNA. Accordingly, analysis of versican transcript poly(A) tail evidenced a longer product 3 h after treatment, but not after 12 h. Versican immunoreaction becomes conspicuous in the superficial stroma only 3 h after E2 injection, whereas the whole stroma is immunoreactive from 6 h onward. These results demonstrate that E2 modulates versican at the transcriptional and posttranscriptional levels in a time-dependent manner.
authors.affiliation: Department of Cell and Developmental Biology, University of São Paulo, Avenida Professor Lineu Prestes, 1524-Room 429, Butantã, São Paulo, SP 05508-000, Brazil. rsalgado@icb.usp.br
authors.forename: Renato M
authors.initials: RM
authors.lastname: Salgado
authors.forename: Ambart C
authors.initials: AC
authors.lastname: Covarrubias
authors.forename: Rodolfo R
authors.initials: RR
authors.lastname: Favaro
authors.forename: Caroline
authors.initials: C
authors.lastname: Serrano-Nascimento
authors.forename: Maria Tereza
authors.initials: MT
authors.lastname: Nunes
authors.forename: Telma M T
authors.initials: TM
authors.lastname: Zorn
mesh_terms: D053959:Alpha-Amanitin / Q000494:pharmacology; D000818:Animals; D004305:Dose-Response Relationship, Drug; D004958:Estradiol / Q000494:pharmacology*; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D051379:Mice; D011061:Poly A; D026723:Polyadenylation / Q000187:drug effects; D012323:RNA Processing, Post-Transcriptional / Q000187:drug effects*; D013997:Time Factors; D014158:Transcription, Genetic / Q000187:drug effects*; D014599:Uterus / Q000187:drug effects* / Q000378:metabolism*; D053675:Versicans / Q000235:genetics* / Q000378:metabolism
chemical_list: D053959:Alpha-Amanitin; D053675:Versicans; D011061:Poly A; D004958:Estradiol