Effect of Dickkopf-1 and colony stimulating factor-2 on the developmental competence, quality, gene expression and live birth rate of buffalo (Bubalus bubalis) embryos produced by hand-made cloning

Shyam, Songyukta; Goel, Prashant; Kumar, Deepak; Malpotra, Shivani; Singh, Manoj Kumar; Lathwal, S. S.; Chand, Subhash; Palta, P.

doi:10.1016/j.theriogenology.2020.07.022

Cited by 20 publications

(14 citation statements)

References 48 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…We found that YY1, SOD1, IL6, G6PD, PCGF2, MCL1, and IFN-τ represented no methylation in the gene regions whereas TIMP2, MTPN, COL4A1, TCF7, AQP9, BMP7, FGF7, WNT2, DKK3, CRABP2, PPARA, DYSF , LEF1 and HDAC8 showed high number of regions with hypermethylation in blastocyst stage cloned embryos compared to their IVF counterparts. Shyam et al 55 also observed a high expression of WNT signalling pathway genes i.e. TCF7 and LEF1 in cloned buffalo blastocyst-stage embryos.…”

Section: Discussionmentioning

confidence: 93%

Global DNA methylation profiles of buffalo (Bubalus bubalis) preimplantation embryos produced by handmade cloning and in vitro fertilization

Malpotra

Goel

Shyam

et al. 2022

Sci Rep

View full text Add to dashboard Cite

Somatic cell nuclear transfer technique (SCNT) has proved to be an outstanding method of multiplication of elite animals but accompanied with low efficiency and live birth rate of cloned animals. Epigenetic alterations of DNA has been one of the culprits behind this issue. Cloned embryos are found to deviate slightly from regular pattern of demethylation and re-methylation at the time of nuclear reprogramming and embryonic development when compared with embryos produced by in vitro fertilization (IVF). Thus, the present study was aimed at evaluating global DNA methylation profiles of cloned embryos at 2-cell, 8-cell and blastocyst stages and compare it with corresponding stages of embryos produced by IVF by using MeDIP-Sequencing on Illumina-based platform. We found out that cloned embryos exhibited significantly different DNA methylation pattern as compared to IVF embryos with respect to distribution of differentially methylated regions in different components of genome, CpG islands distribution and methylation status, gene ontological profiles and pathways affected throughout the developmental stages. The data generated from MeDIP-Seq was validated at blastocyst stage cloned and IVF embryos by bisulfite-sequencing PCR on five randomly selected gene regions.

show abstract

Section: Discussionmentioning

confidence: 93%

Global DNA methylation profiles of buffalo (Bubalus bubalis) preimplantation embryos produced by handmade cloning and in vitro fertilization

Malpotra

Goel

Shyam

et al. 2022

Sci Rep

View full text Add to dashboard Cite

show abstract

“…Results from previous studies from our lab are also in agreement with the data from our present study, viz., Sood et al, performed an RNA-seq study on cloned and IVF blastocysts in which most of the genes related to the Wnt signaling pathway were found to be up-regulated, whereas inhibitors such as DKK-1 were down-regulated, suggesting an altered expression of Wnt signaling pathways in SCNT blastocyst [ 50 ]. Likewise, Shyam et al, reported faulty Wnt signaling in SCNT embryos that was rectified with Dickkopf-1 (a Wnt signaling inhibitor) and CSF-1 treatment in in vitro culture media [ 51 ]. This treatment significantly improved the development competence, quality, and change in gene expression and live birth rate of buffalo embryos.…”

Section: Discussionmentioning

confidence: 99%

Global MicroRNA Expression Profiling of Buffalo (Bubalus bubalis) Embryos at Different Developmental Stages Produced by Somatic Cell Nuclear Transfer and In-Vitro Fertilization Using RNA Sequencing

Goel

Malpotra

Shyam

et al. 2022

Genes

View full text Add to dashboard Cite

Despite the success of cloning technology in the production of offspring across several species, its application on a wide scale is severely limited by the very low offspring rate obtained with cloned embryos. The expression profile of microRNAs (miRNAs) in cloned embryos throughout embryonic development is reported to deviate from regular patterns. The present study is aimed at determining the dynamics of the global expression of miRNA profile in cloned and in-vitro fertilization (IVF) pre-implantation embryos at different developmental stages, i.e., the two-cell, eight-cell, and blastocyst stages, using next-generation sequencing. The results of this study suggest that there is a profound difference in global miRNA profile between cloned and IVF embryos. These differences are manifested throughout the course of embryonic development. The cloned embryos differ from their IVF counterparts in enriched Gene Ontology (GO) terms of biological process, molecular function, cellular component, and protein class categories in terms of the targets of differentially expressed miRNAs. The major pathways related to embryonic development, such as the Wnt signaling pathway, the apoptosis signaling pathway, the FGF signaling pathway, the p53 pathway, etc., were found to be affected in cloned relative to IVF embryos. Overall, these data reveal the distinct miRNA profile of cloned relative to IVF embryos, suggesting that the molecules or pathways affected may play an important role in cloned embryo development.

show abstract

“…Dickkopf-1 (DKK1) is a secretory inhibitor of the canonical WNT signaling pathway. DKK1 supplementation in the IVC media on day 5 of embryo culture could increase blastocyst formation, conception, and birth rates of HMC river buffalo embryos [234]. Genetic instability, including DNA double-strand breaks (DSBs) and chromosome segregation errors, has been found in both cloned human and cloned mouse embryos, resulting in delayed DNA replication and abnormal mitosis [235].…”

Section: Alternative Methods For Cloning Efficiency Improvementmentioning

confidence: 99%

Strategies to Improve the Efficiency of Somatic Cell Nuclear Transfer

Srirattana

Kaneda

Parnpai

2022

IJMS

View full text Add to dashboard Cite

Mammalian oocytes can reprogram differentiated somatic cells into a totipotent state through somatic cell nuclear transfer (SCNT), which is known as cloning. Although many mammalian species have been successfully cloned, the majority of cloned embryos failed to develop to term, resulting in the overall cloning efficiency being still low. There are many factors contributing to the cloning success. Aberrant epigenetic reprogramming is a major cause for the developmental failure of cloned embryos and abnormalities in the cloned offspring. Numerous research groups attempted multiple strategies to technically improve each step of the SCNT procedure and rescue abnormal epigenetic reprogramming by modulating DNA methylation and histone modifications, overexpression or repression of embryonic-related genes, etc. Here, we review the recent approaches for technical SCNT improvement and ameliorating epigenetic modifications in donor cells, oocytes, and cloned embryos in order to enhance cloning efficiency.

show abstract

Effect of Dickkopf-1 and colony stimulating factor-2 on the developmental competence, quality, gene expression and live birth rate of buffalo (Bubalus bubalis) embryos produced by hand-made cloning

Cited by 20 publications

References 48 publications

Global DNA methylation profiles of buffalo (Bubalus bubalis) preimplantation embryos produced by handmade cloning and in vitro fertilization

Global DNA methylation profiles of buffalo (Bubalus bubalis) preimplantation embryos produced by handmade cloning and in vitro fertilization

Global MicroRNA Expression Profiling of Buffalo (Bubalus bubalis) Embryos at Different Developmental Stages Produced by Somatic Cell Nuclear Transfer and In-Vitro Fertilization Using RNA Sequencing

Strategies to Improve the Efficiency of Somatic Cell Nuclear Transfer

Contact Info

Product

Resources

About