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Improved in vitro development of cloned bovine embryos using S-Adenosylhomocysteine, a non-toxic epigenetic modifying reagent.


Citation

Jafari, Shahram and Hosseini, Morteza S. and Hajian, Mahdi and Forouzanfar, Mohsen and Jafarpour, Farnoosh and Abedi, Parvaneh and Ostadhosseini, Somayyeh and Abbasi, Hassan and Gourabi, Hamid and Shahverdi, Abdol H. and Taghi Dizaj, Ahmad Vosough and Anjomshoaa, Maryam and Haron, Abd Wahid and Nordin, Norshariza and Yaakub, Halimatun and Nasr Esfahani, Mohammad Hosein (2011) Improved in vitro development of cloned bovine embryos using S-Adenosylhomocysteine, a non-toxic epigenetic modifying reagent. Molecular Reproduction and Development, 78 (8). pp. 576-584. ISSN 1040-452X; ESSN:1098-2795

Abstract

In this study, fibroblast cells were stably transfected with mouse POU5F1 promoter-driven enhanced green fluorescent protein (EGFP) to investigate the effect of S-adenosylhomocysteine (SAH), the reversible non-toxic inhibitor of DNA-methyltransferases (DNMTs), at different intervals post-fusion on in vitro development of cloned bovine embryos. Treatment with SAH for 12hr resulted in 54.6±7.7% blastocyst production, which was significantly greater than in vitro fertilized embryos (IVF: 37.2±2.7%), cloned embryos treated with SAH for 72hr (31.0±7.6%), and control cloned embryos (34.6±3.6%). The fluorescence intensities of the EGFP-POU5F1 reporter gene at all intervals of SAH treatment, except of 72hr, were significantly higher than control somatic cell nuclear transfers (SCNT) embryos. The intensity of DNA-methylation in cloned embryos treated with SAH for 48hr was similar to that of IVF embryos, and was significantly lower than the other SCNT groups. The levels of H3K9 acetylation in all SCNT groups were significantly lower than IVF embryos. Real-time PCR analysis of gene expression revealed significantly higher expression of POU5F1 in cloned versus IVF blastocysts. Neither embryo production method (SCNT vs. IVF) nor the SAH treatment interval affected expression of the BCL2 gene. Cloned embryos at all intervals of SAH treatment, except for 24hr, had significantly increased VEGF transcript compared to IVF and control SCNT embryos. It was suggested that the time interval of DNMT inhibition may have important consequences on different in vitro features of bovine SCNT, and the improving effects of DNMT inhibition on developmental competency of cloned embryos are restricted to a specific period of time preceding de novo methylation.


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Additional Metadata

Item Type: Article
Divisions: Faculty of Agriculture
DOI Number: https://doi.org/10.1002/mrd.21344
Publisher: Wiley-Blackwell
Keywords: Cloned embryo; Epigenetic; Bovine.
Depositing User: Nur Farahin Ramli
Date Deposited: 28 Nov 2013 02:23
Last Modified: 06 Oct 2015 02:43
Altmetrics: http://www.altmetric.com/details.php?domain=psasir.upm.edu.my&doi=10.1002/mrd.21344
URI: http://psasir.upm.edu.my/id/eprint/23827
Statistic Details: View Download Statistic

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