Estradiol induces transcriptional and posttranscriptional modifications in versican expression in the mouse uterusReport as inadecuate




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Journal of Molecular Histology

, Volume 44, Issue 2, pp 221–229

First Online: 28 December 2012Received: 14 October 2012Accepted: 14 December 2012DOI: 10.1007-s10735-012-9476-1

Cite this article as: Salgado, R.M., Covarrubias, A.C., Favaro, R.R. et al. J Mol Hist 2013 44: 221. doi:10.1007-s10735-012-9476-1

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 polyA 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 polyA 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.

KeywordsEstradiol Versican mRNA stability Mouse uterus PolyA tail  Download fulltext PDF



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