Maryam Salimi1, Abolfazl Shirazi2,3, Mohsen Norouzian2, Ameneh Jafari4, Haleh Edalatkhah2, Maryam Mehravar2, Mohammad Majidi5, Mohammad Mahdi Mehrazar2. 1. Department of Biology and Anatomical Sciences, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran. 2. Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran. 3. Department of Gametes and Cloning, Research Institute of Animal Embryo Technology, Shahrekord University, Shahrekord, Iran. 4. Student Research Committee, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran. 5. Food and Drug Laboratory Research Center, Food and Drug Organization, MOH & ME, Tehran, Iran.
Abstract
BACKGROUND: Currently, the efficient production of chimeric mice and their survival are still challenging. Recent researches have indicated that preimplantation embryo culture media and manipulation lead to abnormal methylation of histone in the H19/Igf2 promotor region and consequently alter their gene expression pattern. This investigation was designed to evaluate the relationship between the methylation state of histone H3 and H19/Igf2 expression in mice chimeric blastocysts. METHODS: Mouse 129/Sv embryonic stem cells (mESCs) expressing the green fluorescent protein (mESCs-GFP) were injected into the perivitelline space of 2.5 days post-coitis (dpc) embryos (C57BL/6) using a micromanipulator. H3K4 and H3K9 methylation, and H19 and Igf2 expression was measured by immunocytochemistry and q-PCR, respectively, in blastocysts. RESULTS: Histone H3 trimethylation in H3K4 and H3K9 in chimeric blastocysts was significantly less and greater, respectively (p< 0.05), than in controls. H19 expression was significantly less (p< 0.05), while Igf2 expression was less, but not significantly so, in chimeric than in control blastocysts. CONCLUSION: Our results showed, that the alteration ofH3K4me3 and H3K9me3 methylation, change H19/Igf2 expression in chimeric blastocysts.
BACKGROUND: Currently, the efficient production of chimeric mice and their survival are still challenging. Recent researches have indicated that preimplantation embryo culture media and manipulation lead to abnormal methylation of histone in the H19/Igf2 promotor region and consequently alter their gene expression pattern. This investigation was designed to evaluate the relationship between the methylation state of histone H3 and H19/Igf2 expression in mice chimeric blastocysts. METHODS: Mouse 129/Sv embryonic stem cells (mESCs) expressing the green fluorescent protein (mESCs-GFP) were injected into the perivitelline space of 2.5 days post-coitis (dpc) embryos (C57BL/6) using a micromanipulator. H3K4 and H3K9 methylation, and H19 and Igf2 expression was measured by immunocytochemistry and q-PCR, respectively, in blastocysts. RESULTS: Histone H3 trimethylation in H3K4 and H3K9 in chimeric blastocysts was significantly less and greater, respectively (p< 0.05), than in controls. H19 expression was significantly less (p< 0.05), while Igf2 expression was less, but not significantly so, in chimeric than in control blastocysts. CONCLUSION: Our results showed, that the alteration ofH3K4me3 and H3K9me3 methylation, change H19/Igf2 expression in chimeric blastocysts.
Authors: William T Poueymirou; Wojtek Auerbach; David Frendewey; Joseph F Hickey; Jennifer M Escaravage; Lakeisha Esau; Anthony T Doré; Sean Stevens; Niels C Adams; Melissa G Dominguez; Nicholas W Gale; George D Yancopoulos; Thomas M DeChiara; David M Valenzuela Journal: Nat Biotechnol Date: 2006-12-24 Impact factor: 54.908
Authors: Gudrun E Moore; Miho Ishida; Charalambos Demetriou; Lara Al-Olabi; Lydia J Leon; Anna C Thomas; Sayeda Abu-Amero; Jennifer M Frost; Jaime L Stafford; Yao Chaoqun; Andrew J Duncan; Rachel Baigel; Marina Brimioulle; Isabel Iglesias-Platas; Sophia Apostolidou; Reena Aggarwal; John C Whittaker; Argyro Syngelaki; Kypros H Nicolaides; Lesley Regan; David Monk; Philip Stanier Journal: Philos Trans R Soc Lond B Biol Sci Date: 2015-03-05 Impact factor: 6.237