Haploidy influences Bak and Bcl-xL mRNA expression and increases incidence of apoptosis in porcine embryos

Jeong, Yu Jeong; Cui, Xiang-Shun; Kim, Bong-Ki; Kim, Il Hwa; Kim, Teoan; Chung, Yon-Bok; Kim, Nam‐Hyung

doi:10.1017/s0967199405003096

Cited by 18 publications

(16 citation statements)

References 15 publications

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“…Interestingly, haploidy increases the incidence of apoptosis in preimplantation embryos, while parthenogenetic activation and parthenogenesis themselves would not lead to apoptosis (Liu et al 2002a, 2002b, Jeong et al 2005. Haploidy is also responsible for diminished blastocyst rates over normal embryos and diploid parthenotes (Henery & Kaufman 1992, Jeong et al 2005. Therefore, the enhanced diploidy we found in parthenotes is consistent with their AI (that was not increased) and improved development rates.…”

Section: Development and Gene Expression In Parthenotessupporting

confidence: 62%

“…Frequency of diploid blastocysts was reported to be lower in parthenotes than in IVF embryos (Wang et al 2008). Interestingly, haploidy increases the incidence of apoptosis in preimplantation embryos, while parthenogenetic activation and parthenogenesis themselves would not lead to apoptosis (Liu et al 2002a, 2002b, Jeong et al 2005. Haploidy is also responsible for diminished blastocyst rates over normal embryos and diploid parthenotes (Henery & Kaufman 1992, Jeong et al 2005.…”

Section: Development and Gene Expression In Parthenotesmentioning

confidence: 99%

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Biological differences between in vitro produced bovine embryos and parthenotes

Gómez¹,

Gutiérrez‐Adán²,

Díez³

et al. 2009

Reproduction

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Parthenotes may represent an alternate ethical source of stem cells, once biological differences between parthenotes and embryos can be understood. In this study, we analyzed development, trophectoderm (TE) differentiation, apoptosis/necrosis, and ploidy in parthenotes and in vitro produced bovine embryos. Subsequently, using real-time PCR, we analyzed the expression of genes expected to underlie the observed differences at the blastocyst stage. In vitro matured oocytes were either fertilized or activated with ionomycin C6-DMAP and cultured in simple medium. Parthenotes showed enhanced blastocyst development and diploidy and reduced TE cell counts. Apoptotic and necrotic indexes did not vary, but parthenotes evidenced a higher relative proportion of apoptotic cells between inner cell mass and TE. The pluripotence-related POU5F1 and the methylation DNMT3A genes were downregulated in parthenotes. Among pregnancy recognition genes, TP-1 was upregulated in parthenotes, while PGRMC1 and PLAC8 did not change. Expression of p66 shc and BAX/BCL2 ratio were higher, and p53 lower, in parthenotes. Among metabolism genes, SLC2A1 was downregulated, while AKR1B1, PTGS2, H6PD, and TXN were upregulated in parthenotes, and SLC2A5 did not differ. Among genes involved in compaction/blastulation, GJA1 was downregulated in parthenotes, but no differences were detected within ATP1A1 and CDH1. Within parthenotes, the expression levels of SLC2A1, TP-1, and H6PD, and possibly AKR1B1, resemble patterns described in female embryos. The pro-apoptotic profile is more pronounced in parthenotes than in embryos, which may differ in their way to channel apoptotic stimuli, through p66 shc and p53respectively, and in their mechanisms to control pluripotency and de novo methylation.Reproduction (2009) 137 285-295

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Section: Development and Gene Expression In Parthenotessupporting

confidence: 62%

Section: Development and Gene Expression In Parthenotesmentioning

confidence: 99%

Biological differences between in vitro produced bovine embryos and parthenotes

Gómez¹,

Gutiérrez‐Adán²,

Díez³

et al. 2009

Reproduction

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“…Nothing is known about the role of BAK1 during early development of the preimplantation bovine embryos; however, BAK1 has been shown to be involved in the regulation of apoptosis during early embryogenesis of the pig [34,35], human [36], and mouse [37], in a developmental stage-specific manner. Although elevated expression of BAK1 has been shown to play a critical role in the induction of apoptosis by inducing permeabilization of the mitochondrial outer membrane and by forming complexes with the protective BCL2-L1 proteins [38], the experimental evidence suggests that post-transcriptional and/or post-translational modification is also involved in regulation of BAK1 activation [39,40].…”

Section: Discussionmentioning

confidence: 99%

“…Accordingly, several studies have shown an important role of BCL2-L1 in the regulation of embryonic cell death in pig [18,34,35,41], mouse [42], human [43], and cattle [44][45][46]. In contrast, some studies have shown that the levels of BCL2-L1 transcripts did not correlated with blastocyst quality [47,48].…”

Section: Discussionmentioning

confidence: 99%

Insulin-like growth factor 2: A modulator of anti-apoptosis related genes (HSP70, BCL2-L1) in bovine preimplantation embryos

et al. 2014

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Intrinsic defects within the embryos, reflected by elevated cell death and low proliferative ability, are considered the most critical factors associated with bovine infertility. The identification of embryonic factors, which are responsible for successful embryo development, is thus critical in designing strategies for infertility intervention. In this experiment, the possible mechanisms involved in both blastomere proliferation and regulation of cell death were studied by analysis of relative expression patterns of IGF-II, BCL2-L1, BAK1, and HSP70 in 3 classes of morphological quality groups (e.g., excellent, good, and poor) of bovine blastocysts produced by IVF. Variation in total blastocyst cell numbers as well as their allocation to inner cell mass and trophectoderm lineages were also determined by differential CDX2 staining. Results showed that transcript levels for IGF-II, BCL2-L1, and the BCL2-L1/BAK1 ratio were higher in excellent- and good-quality blastocysts compared with low-quality blastocysts (P<0.01), whereas mRNA levels for HSP70 were higher in low-quality blastocyst compared with excellent-quality bovine blastocysts (P<0.05). In addition, excellent-quality blastocysts displayed not only greater total cell number but also greater mean inner cell mass/total cell number proportion than that of poor-quality blastocysts (P<0.01). The expression levels of IGF-II showed negative correlation with the levels of HSP70 (r=-0.70; P<0.05); however, the correlation of expression levels of IGF-II with both of BCL2-L1 (r=0.91; P<0.01) and the ratio of BCL2-L1/BAK1 (r=0.78; P<0.05) were highly positive. There was no correlation between the expression levels of IGF-II and BAK1 genes. In conclusion, these observations suggested that levels of endogenous IGF-II transcripts might be associated with the quality of IVF embryos by regulating either apoptosis-related genes or mitogenic actions in bovine preimplantation embryos.

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“…Esta família está subdivididas em dois grupos: Proteínas pró-apoptóticas (Bax, Bad, Bid, Bcl-xS, Bak, Box, Bik, Blk, Bim, HrK, BNIP3), localizadas principalmente na membrana externa da mitocôndria e proteína anti-apoptóticas (Bcl-2, Bcl-xl, Bcl-w, A1, Mcl-1), localizadas no envelope nuclear e no retículo endoplasmático (SPANOS et al, 2002;JEONG et al, 2005). O balanço de proteínas pró e anti-apoptóticas é o que determina se a célula entrará ou não em apoptose (BETTS; KING, 2001;SPANOS et al, 2002).…”

Section: Bcl-2unclassified

Presença da Caspase-3 ativa e das proteínas de reparo de lesões do DNA em embriões suínos ativados partenogeneticamente com alta ou baixa capacidade de desenvolvimento

Coutinho¹

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COUTINHO, A. R. S. Presence of active Caspase-3 and DNA damage and repair proteins in parthenogenetically activated swines embryos of high and low developmental capacity. [Presença de Caspase-3 ativa e proteínas de reparo de lesão do DNA em embriões suínos ativados partenogeneticamente com alta ou baixa capacidade de desenvolvimento]. 2007. 114 f. Tese (Doutorado em Medicina Veterinária) -Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, São Paulo, 2007.Apoptosis is a highly conserved form of cell death that plays a major role in animal development and cellular homeostasis by acting as a quality control mechanism to remove cells that are damaged, nonfunctional, misplaced or supranumerary. This form of cell death plays a major role on preimplantation embryonic development since embryonic cells are often subject to DNA damage, triggering either DNA damage and repair mechanisms or apoptosis. Once initiated the apoptotic process leads to caspase activation and cleavage of cellular substrates. The aim of the present study was to quantify active Caspase-3 (+casp-3) and to determine the role of this pro-apoptotic enzyme on the development of preimplantation porcine embryos, as well as to investigate the presence and localization of DNA damage and repair proteins. Four experiments were conducted using slaughterhouse immature oocytes collected from pre-pubertal gilt ovaries and in vitro matured (IVM) for 44-46 h. Metaphase II (MII) oocytes were parthenogenetically activated (PA) using ionomycin and strontium chloride and cultured in PZM-3 at 38°C, 5% CO 2 in air and high humidity. In the first study embryos were distributed into 4 groups according to cleavage time and cell number: R4cleaved at 24 h with > 4-cells at 48 h; R2 -cleaved at 24 h with 2-3 cells at 48 h; L4 -noncleaved at 24 h with > 4-cells at 48 h; L2 -non-cleaved at 24 h with 2-3 cells at 48 h. Group R embryos showed higher blastocyst rates R4-66.1 (174/263) and R2-59.6 (62/104) and more nuclei per blastocyst, R4-40.1 and R2-38.9, when compared to group L L4-15.4 (6/39) and L2-16.8 (13/77); L4-18.5 and L2-18.3 (Chi-square and Tukey-Kramer, P<0.05); however, there were no differences between R4 and R2 or L4 and L2. The second experiment used only early-(R) and late-cleaved embryos (L), classified at 24 h of IVC, fixed at D-2, D-4 and D-6 and then stained for +casp-3 immunofluorescence. In this embryos fixed at D-2, D-4 and D-6 were considered as sub-experiments conducted in different replicates. Active Caspase-3 cytoplasmic signal was detected in cultured embryos from D2 to D6 and nuclear signal was detected from D5 to D6. The relative amount of immunofluorescence signal for +casp-3 for groups R and L at D2, D4 and D6 of culture was 2.4 vs. 1.4; 1.1 vs. 1.0 and 1.1 vs. 1.2, respectively. A third experiment was conducted to evaluate the role of +casp-3 on porcine preimplantation embryos. In this study the Caspase inhibitor Z-DEVD-fmk (100 uM) was supplemented during maturation of porcine oocytes or embryo culture (D0 to D2 or D4 to D6)....

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Haploidy influences Bak and Bcl-xL mRNA expression and increases incidence of apoptosis in porcine embryos

Cited by 18 publications

References 15 publications

Biological differences between in vitro produced bovine embryos and parthenotes

Biological differences between in vitro produced bovine embryos and parthenotes

Insulin-like growth factor 2: A modulator of anti-apoptosis related genes (HSP70, BCL2-L1) in bovine preimplantation embryos

Presença da Caspase-3 ativa e das proteínas de reparo de lesões do DNA em embriões suínos ativados partenogeneticamente com alta ou baixa capacidade de desenvolvimento

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