Improves the<i>In Vitro</i>Developmental Competence and Reprogramming Efficiency of Cloned Bovine Embryos by Additional Complimentary Cytoplasm

Xu, Lianguang; Mesalam, Ayman; Lee, Kyeong-Lim; Song, Seok-Hwan; Khan, Imran; Chowdhury, M.M.R.; Lv, Wenfa; Kong, Il‐Keun

doi:10.1089/cell.2018.0050

Cited by 24 publications

(25 citation statements)

References 59 publications

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“…Changes in mitochondrial redox state and metabolites have also been associated with changes in histone modification and transcriptome profile (Kopinski et al, 2019). Finally, and further supporting our hypothesis, bovine SCNT blastocysts were recently shown to have a significantly lower mitochondrial mass than expected from the amount of ooplasm removed by enucleation and display increased abundances of DNA methyl transferase (DNMT1 and DNMT3A) transcripts as compared with IVF controls (Xu et al, 2019).…”

Section: Discussionsupporting

confidence: 73%

Mitochondrial DNA Depletion in Granulosa Cell Derived Nuclear Transfer Tissues

Bebbere

Ulbrich

Giller

et al. 2021

Front. Cell Dev. Biol.

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Somatic cell nuclear transfer (SCNT) is a key technology with broad applications that range from production of cloned farm animals to derivation of patient-matched stem cells or production of humanized animal organs for xenotransplantation. However, effects of aberrant epigenetic reprogramming on gene expression compromise cell and organ phenotype, resulting in low success rate of SCNT. Standard SCNT procedures include enucleation of recipient oocytes before the nuclear donor cell is introduced. Enucleation removes not only the spindle apparatus and chromosomes of the oocyte but also the perinuclear, mitochondria rich, ooplasm. Here, we use a Bos taurus SCNT model with in vitro fertilized (IVF) and in vivo conceived controls to demonstrate a ∼50% reduction in mitochondrial DNA (mtDNA) in the liver and skeletal muscle, but not the brain, of SCNT fetuses at day 80 of gestation. In the muscle, we also observed significantly reduced transcript abundances of mtDNA-encoded subunits of the respiratory chain. Importantly, mtDNA content and mtDNA transcript abundances correlate with hepatomegaly and muscle hypertrophy of SCNT fetuses. Expression of selected nuclear-encoded genes pivotal for mtDNA replication was similar to controls, arguing against an indirect epigenetic nuclear reprogramming effect on mtDNA amount. We conclude that mtDNA depletion is a major signature of perturbations after SCNT. We further propose that mitochondrial perturbation in interaction with incomplete nuclear reprogramming drives abnormal epigenetic features and correlated phenotypes, a concept supported by previously reported effects of mtDNA depletion on the epigenome and the pleiotropic phenotypic effects of mtDNA depletion in humans. This provides a novel perspective on the reprogramming process and opens new avenues to improve SCNT protocols for healthy embryo and tissue development.

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Section: Discussionsupporting

confidence: 73%

Mitochondrial DNA Depletion in Granulosa Cell Derived Nuclear Transfer Tissues

Bebbere

Ulbrich

Giller

et al. 2021

Front. Cell Dev. Biol.

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“…The cellular growth arrest occurring in senescence is seemingly regulated through two main pathways, p16 INK4a -Rb and p53-p21 CIP1 (REF 137 ). Importantly, administration of senolytic agents, which selectively eliminate senescence cells, have been shown to improve physical function and increase longevity in mice 138 .…”

Section: Emerging Areas Of Researchmentioning

confidence: 99%

The effects of growth hormone on adipose tissue: old observations, new mechanisms

et al. 2019

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The ability of growth hormone (GH) to induce adipose tissue lipolysis has been known for over five decades; however, the molecular mechanisms that mediate this effect, as well as the ability of GH to inhibit insulin-stimulated glucose uptake, have been scarcely documented. In this same timeframe, our understanding of adipose tissue has evolved to reveal a complex structure with distinct types of adipocytes, depot-specific differences, a biologically significant extracellular matrix and important endocrine properties mediated by adipokines. All of these aforementioned features, in turn, can influence lipolysis. In this Review, we provide a historical and current overview of the lipolytic effect of GH in humans, mice and cultured cells. More globally, we explain lipolysis in terms of GH-induced intracellular signaling and its effect on obesity, insulin resistance and lipotoxicity. In this regard, findings that define molecular mechanisms by which GH induces lipolysis are described. Finally, data are presented for the differential effect of GH on specific adipose tissue depots and on distinct classes of metabolically active adipocytes. Together, these cellular, animal and human studies reveal novel cellular phenotypes and molecular pathways regulating the metabolic effects of GH on adipose tissue.

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“…The zonafree embryos can easily transmit diseases and be damaged during handling (Verma et al, 2015). Recently, the CICT method has been developed to overcome the drawbacks of traditional cloning methods (Xu et al, 2019). Using the CICT method, the cytoplasmic volume of enucleated oocytes is restored by donor-cytoplast oocytes.…”

Section: Figmentioning

confidence: 99%

“…In a previous study, the reconstructed cloned embryo, using CICT, showed that gene expression levels of DNMT 1 and DNMT 3a were significantly lower than in the traditional cloning method, indicating that CICT can improve epigenetic reprogramming. (Xu et al, 2019). In this study, we have evaluated the effect of the cytoplasmic volume on acetylation (acH3K9/14 and acH3K56) and apoptosis.…”

Section: Figmentioning

confidence: 99%

“…Recently, cytoplasmic injection cloning technology (CICT) was developed, in which 30% of the cytoplasm from a donor oocyte is injected into the enucleated recipient oocyte, restoring a normal cytoplasmic volume, and the effects of this process on cloned bovine embryo development have been studied (Xu et al, 2019). Analysis of CICT cloned embryos shows that reprogramming of the nuclei had occurred and the embryo quality was improved, including total cell number (trophectoderm [TE]), genome methylation, and histone deacetylation, in the bovine embryo in comparison with the traditional cloning method.…”

Section: Introductionmentioning

confidence: 99%

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Effect of Additional Cytoplasm of Cloned Embryo on In Vitro Developmental Competence and Reprogramming Efficiency in Mice

Song

et al. 2020

Cellular Reprogramming

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Somatic cell nuclear transfer (SCNT) is an important technique for biological science research. Cytoplasm injection cloning technology (CICT) was developed to improve the reprogramming efficiency as well as to overcome the limitations of SCNT. CICT uses an additional cytoplasm fused with an enucleated oocyte to restore the cytoplasmic volume of the cloned embryo, and this method could improve the reprogramming efficiency of the cloned embryo. In this study, we show that CICT can be adapted to mouse species to overcome the inefficiency of the SCNT method. In this study, results indicate that the two-cell embryo and blastocyst rates of cloned embryos with the use of the CICT method were significantly higher (p < 0.05) than that of the SCNT method (96.6%-1.1% vs. 86.7%-6.0%, 29.5%-2.6% vs. 22.1%-3.0%, respectively). Furthermore, the apoptotic cell number per blastocyst was significantly lower in the CICT group than that in the SCNT group (1.7-0.2 vs. 2.9-0.3, p < 0.05). Moreover, the acH3K9/K14 expression level in the CICT group was greater than that of the SCNT group (p < 0.05), and the relative acH3K56 level in the CICT group was significantly (p < 0.05) higher than that in the SCNT group. These results indicate that CICT helps improve the in vitro developmental competence and quality of cloned embryos.

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Improves theIn VitroDevelopmental Competence and Reprogramming Efficiency of Cloned Bovine Embryos by Additional Complimentary Cytoplasm

Cited by 24 publications

References 59 publications

Mitochondrial DNA Depletion in Granulosa Cell Derived Nuclear Transfer Tissues

Mitochondrial DNA Depletion in Granulosa Cell Derived Nuclear Transfer Tissues

The effects of growth hormone on adipose tissue: old observations, new mechanisms

Effect of Additional Cytoplasm of Cloned Embryo on In Vitro Developmental Competence and Reprogramming Efficiency in Mice

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