Automated Microinjection of Recombinant BCL-X into Mouse Zygotes Enhances Embryo Development

Liu, Xinyu; Fernandes, Roxanne; Gertsenstein, Marina; Perumalsamy, Alagammal; Lai, Ingrid; Chi, Maggie; Moley, Kelle H.; Greenblatt, Ellen; Jurišica, Igor; Casper, Robert F.; Sun, Yu; Jurisicova, Andrea

doi:10.1371/journal.pone.0021687

Cited by 39 publications

(26 citation statements)

References 51 publications

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“…Previous work in bovine embryos identified this adaptor protein as a mediator of permanent embryo arrest [46,67]. In addition, we have recently shown that injection of recBCL2L1 stimulates mitochondrial function in stressed embryos, prevents excessive generation of ROS, and also maintains low levels of phosphorylated p66Shc [52], implicating BCL2 family members as regulators of developmental competence.…”

Section: Discussionmentioning

confidence: 81%

“…We used the cellpermeable antiapoptotic BH4 domain of BCL2L1 (transactivator of transcription [TAT]-BH4) as this peptide can suppress BAX-induced cellular demise [50,51] and can prevent induction of ROS in embryos [52]. However, we found no impact of TAT-BH4 on progression of Nlrp5 tm/tm embryos (Fig.…”

Section: Transactivator Of Transcription-bh4 and Bip Insufficient To mentioning

confidence: 98%

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NLRP5 Mediates Mitochondrial Function in Mouse Oocytes and Embryos1

Fernandes

Tsuda

Perumalsamy

et al. 2012

Biology of Reproduction

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Unraveling molecular pathways responsible for regulation of early embryonic development is crucial for our understanding of female infertility. Maternal determinants that control the transition from oocyte to embryo are crucial molecules that govern developmental competence of the newly conceived zygote. We describe a series of defects that are triggered by a disruption of maternal lethal effect gene, Nlrp5. Previous studies have shown that Nlrp5 hypomorph embryos fail to develop beyond the two-cell stage. Despite its importance in preimplantation development, the mechanism by which the embryo arrest occurs remains unclear. We confirmed that Nlrp5 mutant and wild-type females possess comparable ovarian germ pool and follicular recruitment rates. However, ovulated oocytes lacking Nlrp5 have abnormal mitochondrial localization and increased activity in order to sustain physiological ATP content. This results in an accumulation of reactive oxygen species and increased cellular stress causing mitochondrial depletion. Compromised cellular state is also accompanied by increased expression of cell death inducer Bax and depletion of cytochrome c. However, neither genetic deletion (Bax/Nlrp5 double knockout) nor mimetic interference (BH4 domain or Bax inhibitory peptide) were sufficient to alleviate embryo demise caused by depletion of Nlrp5. We therefore conclude that lack of Nlrp5 in oocytes triggers premature activation of the mitochondrial pool, causing mitochondrial damage that cannot be rescued by inactivation of Bax.

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Section: Discussionmentioning

confidence: 81%

Section: Transactivator Of Transcription-bh4 and Bip Insufficient To mentioning

confidence: 98%

NLRP5 Mediates Mitochondrial Function in Mouse Oocytes and Embryos1

Fernandes

Tsuda

Perumalsamy

et al. 2012

Biology of Reproduction

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“…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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“…It was demonstrated that, when the expression of HDAC1 is inhibited, expression of BCLXL goes down and results in apoptosis [51]. Studies conducted on mouse preimplantation embryos have also demonstrated the crucial role of BCLXL in protecting the embryos to enter ZGA phase [28,29]. Even though it is yet to be established in rabbit embryos, it is safe to assume that the embryo relies on BCLXL and HDAC1 expression to achieve ZGA.…”

Section: Discussionmentioning

confidence: 99%

“…Among which the BAX/BCLXL ratio indicates survival or death of a cell [26,27]. BCLXL was required for cell survival and development through the ZGA phase in murine preimplantation embryos and the microinjection of BCLXL could rescue embryos and aid in ZGA [28,29]. It was also shown that BCLXL has anti-proliferative effects along with anti-apoptotic properties in murine cancer cell lines [30].…”

Section: Introductionmentioning

confidence: 99%

Candidate gene expression patterns in rabbit preimplantation embryos developed in vivo and in vitro

Gibence

Brahmasani

Mahesh

et al. 2014

J Assist Reprod Genet

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Purpose The levels and timing of expression of genes like BCLXL, HDAC1 and pluripotency marker genes namely, OCT4, SOX2, NANOG and KLF4 are known to influence preimplantation embryo development. Despite this information, precise understanding of their influence during preimplantation embryo development is lacking. The present study attempts to compare the expression of these genes in the in vivo and in vitro developed preimplantation embryos. Methods The in vivo and in vitro developed rabbit embryos collected at distinct developmental stages namely, pronuclear, 2 cell, 4 cell, 8 cell, 16 cell, Morula and blastocyst were compared at the transcriptional and translational levels using Real Time PCR and immunocytochemical studies respectively. Results The study establishes the altered levels of candidate genes at the transcriptional level and translational level with reference to the zygotic genome activation (ZGA) phase of embryo development in the in vivo and in vitro developed embryos. The expression of OCT4, KLF4, NANOG and SOX2 genes were higher in the in vitro developed embryos whereas and HDAC1 was lower. BCLXL expression had its peak at ZGA in in vivo developed embryos. Protein expression of all the candidate genes was observed in the embryos. BCLXL, KLF4 and NANOG exhibited diffused localisation whereas HDAC1, OCT4, and SOX2 exhibited nuclear localisation. Conclusions This study leads to conclude that BCLXL peak expression at the ZGA phase may be a requirement for embryo development. Further expression of all the candidate genes was influenced by ZGA phase of development at the transcript level, but not at the protein level.

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Automated Microinjection of Recombinant BCL-X into Mouse Zygotes Enhances Embryo Development

Cited by 39 publications

References 51 publications

NLRP5 Mediates Mitochondrial Function in Mouse Oocytes and Embryos1

NLRP5 Mediates Mitochondrial Function in Mouse Oocytes and Embryos1

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

Candidate gene expression patterns in rabbit preimplantation embryos developed in vivo and in vitro

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