S-adenosylhomocysteine treatment of adult female fibroblasts alters X-chromosome inactivation and improves in vitro embryo development after somatic cell nuclear transfer

Jeon, Byeong‐Gyun; Coppola, G.; Perrault, Steven D.; Rho, Gyu‐Jin; Betts, Dean H.; King, W. A.

doi:10.1530/rep-07-0442

Cited by 47 publications

(57 citation statements)

References 75 publications

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“…Therefore, treating donor cells with pharmacological agents to remove some epigenetic marks before NT may improve the ability of the donor cells to be fully reprogrammed by the recipient ooplasm. Pretreatment of bovine fibroblast donor cells by DNA demethylation agents such as 5-aza-2Ј-deoxycytidine or S-adenosyl homocysteine has been shown to decrease the global methylation levels in the donor cells but provide little or no improvement on the blastocyst development of embryos reconstructed from the treated cells (9,10). Histone-deacetylase inhibitors (HDACi) such as trichostatin A (TSA) enhance the pool of acetylated histones (11).…”

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confidence: 99%

Somatic Nucleus Reprogramming Is Significantly Improved by m-Carboxycinnamic Acid Bishydroxamide, a Histone Deacetylase Inhibitor

Dai

Hao

Hou

et al. 2010

Journal of Biological Chemistry

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Somatic cell nuclear transfer (SCNT) has shown tremendous potential for understanding the mechanisms of reprogramming and creating applications in the realms of agriculture, therapeutics, and regenerative medicine, although the efficiency of reprogramming is still low. Somatic nucleus reprogramming is triggered in the short time after transfer into recipient cytoplasm, and therefore, this period is regarded as a key stage for optimizing SCNT. Here we report that CBHA, a histone deacetylase inhibitor, modifies the acetylation status of somatic nuclei and increases the developmental potential of mouse cloned embryos to reach pre-and post-implantation stages. Furthermore, the cloned embryos treated by CBHA displayed higher efficiency in the derivation of nuclear transfer embryonic stem cell lines by promoting outgrowths. More importantly, CBHA increased blastocyst quality compared with trichostatin A, another prevalent histone deacetylase inhibitor reported previously. Use of CBHA should improve the productivity of SCNT for a variety of research and clinical applications, and comparisons of cells with different levels of pluripotency and treated with CBHA versus trichostatin A will facilitate studies of the mechanisms of reprogramming.Reprogramming of a terminally differentiated nucleus is successfully achieved in many species through somatic cell nuclear transfer (SCNT) 3 technology. However, its efficiency remains very low, limiting its application in agriculture to well bred livestock propagation and in species preservation (1) and regenerative medicine. The reprogramming process remains largely unknown; at the time of its transfer into an enucleated oocyte the somatic donor nucleus is in a configuration very different from a germ cell or an embryonic nucleus. Its own specific program of gene expression will have to be turned on, and the specific epigenetic modifications and chromatin configuration were erased (2). Extensive chromatin remodeling takes place as soon as the somatic nucleus is in contact with the oocyte cytoplasm, and two main types of epigenetic marks are directly involved in gene expression regulation; that is, methylation of histones or DNA and acetylation of histones (3-5). During normal pre-implantation development, the embryonic genome is progressively demethylated up to the early blastocyst stage. This is followed by differential remethylation in the two lineages of the blastocyst, the pluripotent inner cell mass and the trophoblast (6), but this process has been shown to be errorprone in SCNT embryos (7,8).Attempts to improve reprogramming during nuclear transfer have, therefore, focused primarily on modifying the epigenetic configuration of the donor nuclei before nuclear transfer. Therefore, treating donor cells with pharmacological agents to remove some epigenetic marks before NT may improve the ability of the donor cells to be fully reprogrammed by the recipient ooplasm. Pretreatment of bovine fibroblast donor cells by DNA demethylation agents such as 5-aza-2Ј-deoxycytidine or S-adenosy...

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confidence: 99%

Somatic Nucleus Reprogramming Is Significantly Improved by m-Carboxycinnamic Acid Bishydroxamide, a Histone Deacetylase Inhibitor

Dai

Hao

Hou

et al. 2010

Journal of Biological Chemistry

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“…We adapted this QC reaction from a method described elsewhere (38) for use in a SYBR green assay format. The LSG-qPCR and QC-qPCR were separate reactions (in separate PCR tubes) in which the reaction mixtures were prepared by use of the same chemistry and cycling parameters.…”

Section: Methodsmentioning

confidence: 99%

Detection and Differentiation of Leishmania spp. in Clinical Specimens by Use of a SYBR Green-Based Real-Time PCR Assay

et al. 2017

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Leishmaniasis in humans is caused by Leishmania spp. in the subgenera Leishmania and Viannia. Species identification often has clinical relevance. Until recently, our laboratory relied on conventional PCR amplification of the internal transcribed spacer 2 (ITS2) region (ITS2-PCR) followed by sequencing analysis of the PCR product to differentiate Leishmania spp. Here we describe a novel real-time quantitative PCR (qPCR) approach based on the SYBR green technology (LSG-qPCR), which uses genus-specific primers that target the ITS1 region and amplify DNA from at least 10 Leishmania spp., followed by analysis of the melting temperature (T m ) of the amplicons on qPCR platforms (the Mx3000P qPCR system [Stratagene-Agilent] and the 7500 real-time PCR system [ABI Life Technologies]). We initially evaluated the assay by testing reference Leishmania isolates and comparing the results with those from the conventional ITS2-PCR approach. Then we compared the results from the real-time and conventional molecular approaches for clinical specimens from 1,051 patients submitted to the reference laboratory of the Centers for Disease Control and Prevention for Leishmania diagnostic testing. Specimens from 477 patients tested positive for Leishmania spp. with the LSG-qPCR assay, specimens from 465 of these 477 patients also tested positive with the conventional ITS2-PCR approach, and specimens from 10 of these 465 patients had positive results because of retesting prompted by LSG-qPCR positivity. On the basis of the T m values of the LSG-qPCR amplicons from reference and clinical specimens, we were able to differentiate four groups of Leishmania parasites: the Viannia subgenus in aggregate; the Leishmania

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“…In vitro cultured fibroblast cells, which had been derived from the tissue of foetuses and adult specimens, or derived from the follicle (granulose cells/cumulus cells) are the commonly used source of nuclear donor cells in the somatic cell cloning of mammals (Skrzyszowska et al, 2008, Jeon et al 2008, Van Thuan et al 2009, Dai et al 2010, Samiec and Skrzyszowska 2010a, 2012a. The degree of molecular and epigenetic differentiation of these cells, which is related both to the advanced methylation profile of DNA cytosine residues and to the lysine deacetylation profile of histones forming nucleosomal core of nuclear chromatin, often seems to make impossible converting the covalent modifications back to a totipotent state of embryonic (zygotic) cells.…”

Section: The Use Of Mscs In Mammals Scnt -Epigenetic Aspectmentioning

confidence: 99%

Characterization of mesenchymal stem cells and their application in experimental embryology

Opiela¹,

Samiec²

2013

Polish Journal of Veterinary Sciences

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The efficiency of somatic cell cloning (somatic cell nuclear transfer; SCNT) as well as in vitro fertilization/in vitro embryo production (IVF/IVP) in mammals stay at relatively same level for over a decade. Despite plenty of different approaches none satisfactory break-through took place. In this article, we briefly summarize the implementation of mesenchymal stem cells (MSCs) for experimental embryology. The advantages of using MSCs as nuclear donors in somatic cell cloning and in vitro embryo culture are described. The description of results obtained with these cells in mammalian embryo genomic engineering is presented.

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S-adenosylhomocysteine treatment of adult female fibroblasts alters X-chromosome inactivation and improves in vitro embryo development after somatic cell nuclear transfer

Cited by 47 publications

References 75 publications

Somatic Nucleus Reprogramming Is Significantly Improved by m-Carboxycinnamic Acid Bishydroxamide, a Histone Deacetylase Inhibitor

Somatic Nucleus Reprogramming Is Significantly Improved by m-Carboxycinnamic Acid Bishydroxamide, a Histone Deacetylase Inhibitor

Detection and Differentiation of Leishmania spp. in Clinical Specimens by Use of a SYBR Green-Based Real-Time PCR Assay

Characterization of mesenchymal stem cells and their application in experimental embryology

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