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Suppression of the transcription factor MSX1 gene delays bovine preimplantation embryo development in vitro
D Tesfaye1, A Regassa, F Rings
1Animal Breeding and Husbandry Group, Institute of Animal Science, University of Bonn, Endenicher Allee 15, 53115 Bonn, Germany. tesfaye@itw.uni-bonn.de
Abstract:
This study was conducted to investigate the effect of suppressing transcription factor gene MSX1 on the development of in vitro produced bovine oocytes and embryos, and identify its potential target genes regulated by this gene. Injection of long double-stranded RNA (LdsRNA) and small interfering RNA (siRNA) at germinal vesicle stage oocyte reduced MSX1 mRNA expression by 73 and 37% respectively at metaphase II stage compared with non-injected controls. Similarly, injection of the same anti-sense oligomers at zygote stage reduced MSX1 mRNA expression by 52 and 33% at 8-cell stage compared with non-injected controls. Protein expression was also reduced in LdsRNA- and siRNA-injected groups compared with non-injected controls at both stages. Blastocysts rates were 33, 28, 20 and 18% in non-injected control, scrambled RNA (scRNA), LdsRNA- and siRNA-injected groups respectively. Cleavage rates were also significantly reduced in Smartpool siRNA (SpsiRNA)-injected group (53.76%) compared with scRNA-injected group (57.76%) and non-injected control group (61%). Large-scale gene expression analysis showed that 135 genes were differentially regulated in SpsiRNA-injected group compared with non-injected controls, of which 54 and 81 were down- and up-regulated respectively due to suppression of MSX1. Additionally, sequence homology mapping and gene enrichment analysis with known human pathway information identified several functional modules that were affected due to suppression of MSX1. In conclusion, suppression of MSX1 affects oocyte maturation, embryo cleavage rate and the expression of several genes, suggesting its potential role in the development of bovine preimplantation embryos.
Insights
Suppressing the transcription factor gene MSX1 significantly impacts bovine oocyte maturation and early embryo development. This gene
Area of Science:
- Reproductive Biology
- Developmental Biology
- Molecular Genetics
Background:
- The transcription factor MSX1 plays a role in embryonic development, but its specific function in bovine oocytes and preimplantation embryos is not fully understood.
- Understanding gene regulation is crucial for improving in vitro embryo production (IVEP) efficiency in cattle.
Purpose of the Study:
- To investigate the effect of suppressing MSX1 gene expression on the development of in vitro produced bovine oocytes and embryos.
- To identify potential target genes regulated by MSX1 during bovine preimplantation development.
Main Methods:
- Messenger RNA (mRNA) and protein expression of MSX1 were suppressed in bovine oocytes and zygotes using long double-stranded RNA (LdsRNA) and small interfering RNA (siRNA).
- Oocyte maturation, embryo cleavage rates, and blastocyst formation rates were assessed.
- Large-scale gene expression analysis (RNA sequencing) and bioinformatics tools were employed to identify differentially regulated genes and affected pathways.
Main Results:
- MSX1 suppression significantly reduced MSX1 mRNA and protein expression in oocytes and early embryos.
- Blastocyst rates decreased in LdsRNA- and siRNA-injected groups compared to controls.
- Cleavage rates were significantly reduced in the Smartpool siRNA (SpsiRNA)-injected group.
- Gene expression analysis revealed 135 differentially regulated genes (54 down-regulated, 81 up-regulated) upon MSX1 suppression.
Conclusions:
- Suppression of MSX1 negatively affects bovine oocyte maturation and embryo cleavage rates.
- MSX1 plays a critical role in regulating gene expression during bovine preimplantation development.
- These findings suggest MSX1 is a key factor in bovine early embryonic development.
