Phosphorylated histone H2A.x in porcine embryos produced by IVF and somatic cell nuclear transfer

Bohrer, Rodrigo Camponogara; Che, Limei; Gonçalves, Paulo Bayard Dias; Duggavathi, Raj; Bordignon, Vilceu

doi:10.1530/rep-13-0271

Cited by 19 publications

(19 citation statements)

References 55 publications

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“…These results are in agreement with previous reports of higher blastocyst rates among fast-developing embryos [19,8]. Slow-cleaving embryos might have a higher incidence of chromosomal abnormalities, altered gene expression and increased DNA double-strand breaks compared to rapidly cleaving embryos [8,7]. In contrast, there are reports of abnormalities in the expression of imprinted genes in fast-developing mice embryos [20].…”

Section: Discussionsupporting

confidence: 92%

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Developmental Block and Programmed Cell Death in Bos indicus Embryos: Effects of Protein Supplementation Source and Developmental Kinetics

et al. 2015

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The aims of this study were to determine if the protein source of the medium influences zebu embryo development and if developmental kinetics, developmental block and programmed cell death are related. The culture medium was supplemented with either fetal calf serum or bovine serum albumin. The embryos were classified as Fast (n = 1,235) or Slow (n = 485) based on the time required to reach the fourth cell cycle (48 h and 90 h post insemination - hpi -, respectively). The Slow group was further separated into two groups: those presenting exactly 4 cells at 48 hpi (Slow/4 cells) and those that reached the fourth cell cycle at 90 hpi (Slow). Blastocyst quality, DNA fragmentation, mitochondrial membrane potential and signs of apoptosis or necrosis were evaluated. The Slow group had higher incidence of developmental block than the Fast group. The embryos supplemented with fetal calf serum had lower quality. DNA fragmentation and mitochondrial membrane potential were absent in embryos at 48 hpi but present at 90 hpi. Early signs of apoptosis were more frequent in the Slow and Slow/4 cell groups than in the Fast group. We concluded that fetal calf serum reduces blastocyst development and quality, but the mechanism appears to be independent of DNA fragmentation. The apoptotic cells detected at 48 hpi reveal a possible mechanism of programmed cell death activation prior to genome activation. The apoptotic cells observed in the slow-developing embryos suggested a relationship between programmed cell death and embryonic developmental kinetics in zebu in vitro-produced embryos.

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Section: Discussionsupporting

confidence: 92%

“…Embryos that fail to accomplish this task do not survive beyond the eight-cell stage; this phenomenon is known as developmental block. At this stage, a low level of DNA fragmentation [7] and rapid development [8] are indicative of embryonic quality.…”

Section: Introductionmentioning

confidence: 99%

Developmental Block and Programmed Cell Death in Bos indicus Embryos: Effects of Protein Supplementation Source and Developmental Kinetics

et al. 2015

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“…To support the role of H2AX in embryogenesis, H2AX presence during embryonic development was also documented in other species, including Xenopus laevis (73), porcine embryos (74) and different human ESC lines (75). …”

Section: The Histone H2ax Non-canonical Rolesmentioning

confidence: 99%

Multiple facets of histone variant H2AX: a DNA double-strand-break marker with several biological functions

Turinetto

Giachino

2015

Nucleic Acids Research

305

221

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In the last decade, many papers highlighted that the histone variant H2AX and its phosphorylation on Ser 139 (γH2AX) cannot be simply considered a specific DNA double-strand-break (DSB) marker with a role restricted to the DNA damage response, but rather as a ‘protagonist’ in different scenarios. This review will present and discuss an up-to-date view regarding the ‘non-canonical’ H2AX roles, focusing in particular on possible functional and structural parts in contexts different from the canonical DNA DSB response. We will present aspects concerning sex chromosome inactivation in male germ cells, X inactivation in female somatic cells and mitosis, but will also focus on the more recent studies regarding embryonic and neural stem cell development, asymmetric sister chromosome segregation in stem cells and cellular senescence maintenance. We will discuss whether in these new contexts there might be a relation with the canonical DNA DSB signalling function that could justify γH2AX formation. The authors will emphasize that, just as H2AX phosphorylation signals chromatin alteration and serves the canonical function of recruiting DSB repair factors, so the modification of H2AX in contexts other than the DNA damage response may contribute towards creating a specific chromatin structure frame allowing ‘non-canonical’ functions to be carried out in different cell types.

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“…The number and size of H2AX139ph foci were evaluated in each nucleus using the SimplePCI Imaging Software (Compix, Inc), and foci lager than 0.3 µm 3 were counted as a site of DSBs. 47 All RAD51 and 53BP1 foci colocalized with H2AX139ph foci were also counted.…”

Section: Downloaded By [Northwestern University] At 16:21 03 Februarymentioning

confidence: 99%

“…47 In somatic cells, histone modifications play important roles in DSB repair. For instance, the acetylation of histone H4K16 is critical for DNA damage response and DSB repair by HR and NHEJ pathways.…”

Section: Introductionmentioning

confidence: 99%

Inhibition of histone deacetylases enhances DNA damage repair in SCNT embryos

2014

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Recent studies have shown that DNA damage affects embryo development and also somatic cell reprogramming into induced pluripotent stem (iPS) cells. It has been also shown that treatment with histone deacetylase inhibitors (HDACi) improves development of embryos produced by somatic cell nuclear transfer (SCNT) and enhances somatic cell reprogramming. There is evidence that increasing histone acetylation at the sites of DNA double-strand breaks (DSBs) is critical for DNA damage repair. Therefore, we hypothesized that HDACi treatment enhances cell programming and embryo development by facilitating DNA damage repair. To test this hypothesis, we first established a DNA damage model wherein exposure of nuclear donor cells to ultraviolet (UV) light prior to nuclear transfer reduced the development of SCNT embryos proportional to the length of UV exposure. Detection of phosphorylated histone H2A.x (H2AX139ph) foci confirmed that exposure of nuclear donor cells to UV light for 10 s was sufficient to increase DSBs in SCNT embryos. Treatment with HDACi during embryo culture increased development and reduced DSBs in SCNT embryos produced from UV-treated cells. Transcript abundance of genes involved in either the homologous recombination (HR) or nonhomologous end-joining (NHEJ) pathways for DSBs repair was reduced by HDACi treatment in developing embryos at day 5 after SCNT. Interestingly, expression of HR and NHEJ genes was similar between HDACi-treated and control SCNT embryos that developed to the blastocyst stage. This suggested that the increased number of embryos that could achieve the blastocyst stage in response to HDACi treatment have repaired DNA damage. These results demonstrate that DNA damage in nuclear donor cells is an important component affecting development of SCNT embryos, and that HDACi treatment after nuclear transfer enhances DSBs repair and development of SCNT embryos.

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Phosphorylated histone H2A.x in porcine embryos produced by IVF and somatic cell nuclear transfer

Cited by 19 publications

References 55 publications

Developmental Block and Programmed Cell Death in Bos indicus Embryos: Effects of Protein Supplementation Source and Developmental Kinetics

Developmental Block and Programmed Cell Death in Bos indicus Embryos: Effects of Protein Supplementation Source and Developmental Kinetics

Multiple facets of histone variant H2AX: a DNA double-strand-break marker with several biological functions

Inhibition of histone deacetylases enhances DNA damage repair in SCNT embryos

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