Author:
Oliveira Clara Slade,Saraiva Naiara Zoccal,Cruz Maria Helena Coelho,Mazeti Bruna,Oliveira Leticia Zoccolaro,Lopes Flavia Lombardi,Garcia Joaquim Mansano
Abstract
During initial development, both X chromosomes are active in females, and one of them must be silenced at the appropriate time in order to dosage compensate their gene expression levels to male counterparts. Silencing involves epigenetic mechanisms, including histone deacetylation. Major X chromosome inactivation (XCI) in bovine occurs between hatching and implantation, althoughin vitroculture conditions might disrupt the silencing process, increasing or decreasing X-linked gene expression. In this study, we aimed to address the roles of histone deacetylase inhibition by trichostatin A (TSA) on female preimplantation development. We tested the hypothesis that by enhancing histone acetylation, TSA would increase the percentage of embryos achieving 16-cell stage, reducing percentage of embryos blocked at 8-cell stage, and interfere with XCI in IVF embryos. We noticed that after TSA treatment, acetylation levels in individual blastomeres of 8–16 cell embryos were increased twofold on treated embryos, and the same was detected for blastocysts. Changes among blastomere levels within the same embryo were diminished on TSA group, as low-acetylated blastomeres were no longer detected. The percentage of embryos that reached the 5th cleavage cycle 118 h after IVF, analyzed by Hoechst staining, remained unaltered after TSA treatment. Then, we assessedXISTandG6PDexpression in individual female bovine blastocysts by quantitative real-time PCR. Even thoughG6PDexpression remained unaltered after TSA exposure,XISTexpression was eightfold decreased, and we also detected a major decrease in the percentage of blastocysts expressing detectableXISTlevels after TSA treatment. Based on these results, we conclude that HDAC is involved on XCI process in bovine embryos, and its inhibition might delay X chromosome silencing and attenuate aberrantXISTexpression described for IVF embryos.
Subject
Cell Biology,Obstetrics and Gynaecology,Endocrinology,Embryology,Reproductive Medicine
Cited by
17 articles.
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