Imprinted gene expression in in vivo- and in vitro-produced bovine embryos and chorio-allantoic membranes

Perecin, Felipe; Méo, Simone Cristina; Yamazaki, W.; Merighe, Giovana Krempel Fonseca; Meirelles, Flávio Vieira; Garcia, Joaquim Mansano

doi:10.4238/vol8-1gmr541

Cited by 23 publications

(16 citation statements)

References 37 publications

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“…This imprinting mechanism is important to determine the parent of origin expression of a particular group of genes called imprinted genes, which will be responsible for extraembryonic and embryonic tissue development. In cloned embryos, a considerable number of studies have reported deficient epigenetic reprogramming, where abnormal patterns of DNA methylation [11][12][13] and imprinted gene expression [13][14][15][16] were found. Because some of these imprinted genes, such as IGF2 and H19, are essential for cell differentiation and tissue growth, this raised the question as to whether epigenetic failure could account for abnormal embryo development and early loss in SCNT pregnancies.…”

Section: Introductionmentioning

confidence: 99%

Loss of Methylation at H19 DMD Is Associated with Biallelic Expression and Reduced Development in Cattle Derived by Somatic Cell Nuclear Transfer1

Suzuki

Therrien

Filion

et al. 2011

Biology of Reproduction

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Although cloning of mammals has been achieved successfully, the percentage of live offspring is very low because of reduced fetal size and fewer implantation sites. Recent studies have attributed such pathological conditions to abnormal reprogramming of the donor cell used for cloning. The inability of the oocyte to fully restore the differentiated status of a somatic cell to its pluripotent and undifferentiated state is normally evidenced by aberrant DNA methylation patterns established throughout the genome during development to blastocyst. These aberrant methylation patterns are associated with abnormal expression of imprinted genes, which among other genes are essential for normal embryo development and gestation. We hypothesized that embryo loss and low implantation rates in cattle derived by somatic cell nuclear transfer (SCNT) are caused by abnormal epigenetic reprogramming of imprinted genes. To verify our hypothesis, we analyzed the parental expression and the differentially methylated domain (DMD) methylation status of the H19 gene. Using a parental-specific analysis, we confirmed for the first time that H19 biallelic expression is tightly associated with a severe demethylation of the paternal H19 DMD in SCNT embryos, suggesting that these epigenetic anomalies to the H19 locus could be directly responsible for the reduced size and low implantation rates of cloned embryos in cattle.

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Section: Introductionmentioning

confidence: 99%

Loss of Methylation at H19 DMD Is Associated with Biallelic Expression and Reduced Development in Cattle Derived by Somatic Cell Nuclear Transfer1

Suzuki

Therrien

Filion

et al. 2011

Biology of Reproduction

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“…The gene expression of embryos and fetuses produced by normal fertilization with sperm in comparison with embryos produced by NT shows numerous differences (Wrenzycki et al, 2001;Nowak-Imialek et al, 2008;Perecin et al, 2009). Conservation of appropriate DNA methylation patterns appears to be a problem in NT embryos (Bourc'his et al, 2001;Kang et al, 2002;Cezar et al, 2003;Santos et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

Proteomic analysis of the major cellular proteins of bovine trophectoderm cell lines derived from IVP, parthenogenetic and nuclear transfer embryos: Reduced expression of annexins I and II in nuclear transfer-derived cell lines

Talbot¹,

Powell²,

Caperna³

et al. 2010

Animal Reproduction Science

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M.; Caperna, Thomas J.; and Garrett, Wesley M., "Proteomic analysis of the major cellular proteins of bovine trophectoderm cell lines derived from IVP, parthenogenetic and nuclear transfer embryos: Reduced expression of annexins I and II in nuclear transfer-derived cell lines" (2010 Trophectoderm cell lines were established from 8-day in vitro-cultured embryos of cattle derived from fertilization (IVF), somatic cell nuclear transfer (NT), or parthenogenetic activation (P) of in vitro-matured oocytes and from five 8-day-old in vivo (V) embryos. The most abundant cellular proteins of 5 V-, 16 NT-, 12 P-, and 16 IVF-derived cell lines were compared by 2D-gel electrophoresis and mass spectrometry; that is, the unaltered thiourea/urea extract of each cell culture was analyzed. Common protein spots (n = 118) were examined, and 95% were identified with significant scores from protein and gene database searches. Of the proteins detected and identified, actin and cytokeratin-8 were found to be the most abundant. Other prominent cellular proteins were metabolic enzymes such as aldose reductase, phosphoglycerate mutase, enolase, triosephosphate isomerase, cytoskeletal interacting proteins transgelin and stratifin, anti-oxidant proteins peroxiredoxin 1 and anti-oxidant protein 2, and the calcium-dependent lipid-binding proteins annexins I and II. In comparative analysis of the 2D-gels, the NT-derived trophectoderm had less annexins I and II in comparison to the IVF-and P-derived trophectoderm. Because annexins I and II are abundant in the placenta and have functions important to the maintenance of placentation, the down-regulation of the annexin genes in the cultured NT trophectoderm may be related to the frequent failures of NT pregnancies. Published by Elsevier B.V.

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“…With respect to IGF2R , this gene plays an important role as a negative effector in fetal growth since it promotes IGF2 arrest into lysosomes followed by degradation [59], [60]. It has been hypothesized that dysregulation of imprinted genes such as IGF2R are the cause of poor developmental rates observed in SCNT [56], and alterations of IGF2R expression has often been associated to common problems in cloned embryos [61]. For instance, in sheep, parthenogenetic embryos that express low levels of IGF2 and high levels of IGF2R and H19 , have retarded growth when compared to control embryos [62]–[64].…”

Section: Discussionmentioning

confidence: 99%

Development to Term of Cloned Cattle Derived from Donor Cells Treated with Valproic Acid

Sangalli

Chiaratti

et al. 2014

PLoS ONE

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Cloning of mammals by somatic cell nuclear transfer (SCNT) is still plagued by low efficiency. The epigenetic modifications established during cellular differentiation are a major factor determining this low efficiency as they act as epigenetic barriers restricting reprogramming of somatic nuclei. In this regard, most factors that promote chromatin decondensation, including histone deacetylase inhibitors (HDACis), have been found to increase nuclear reprogramming efficiency, making their use common to improve SCNT rates. Herein we used valproic acid (VPA) in SCNT to test whether the treatment of nuclear donor cells with this HDACi improves pre- and post-implantation development of cloned cattle. We found that the treatment of fibroblasts with VPA increased histone acetylation without affecting DNA methylation. Moreover, the treatment with VPA resulted in increased expression of IGF2R and PPARGC1A, but not of POU5F1. However, when treated cells were used as nuclear donors no difference of histone acetylation was found after oocyte reconstruction compared to the use of untreated cells. Moreover, shortly after artificial activation the histone acetylation levels were decreased in the embryos produced with VPA-treated cells. With respect to developmental rates, the use of treated cells as donors resulted in no difference during pre- and post-implantation development. In total, five clones developed to term; three produced with untreated cells and two with VPA-treated cells. Among the calves from treated group, one stillborn calf was delivered at day 270 of gestation whereas the other one was delivered at term but died shortly after birth. Among the calves from the control group, one died seven days after birth whereas the other two are still alive and healthy. Altogether, these results show that in spite of the alterations in fibroblasts resulting from the treatment with VPA, their use as donor cells in SCNT did not improve pre- and post-implantation development of cloned cattle.

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Imprinted gene expression in in vivo- and in vitro-produced bovine embryos and chorio-allantoic membranes

Cited by 23 publications

References 37 publications

Loss of Methylation at H19 DMD Is Associated with Biallelic Expression and Reduced Development in Cattle Derived by Somatic Cell Nuclear Transfer1

Loss of Methylation at H19 DMD Is Associated with Biallelic Expression and Reduced Development in Cattle Derived by Somatic Cell Nuclear Transfer1

Proteomic analysis of the major cellular proteins of bovine trophectoderm cell lines derived from IVP, parthenogenetic and nuclear transfer embryos: Reduced expression of annexins I and II in nuclear transfer-derived cell lines

Development to Term of Cloned Cattle Derived from Donor Cells Treated with Valproic Acid

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