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Canonical WNT signaling regulates development of bovine embryos to the blastocyst stage.

Denicol AC, Dobbs KB, McLean KM, Carambula SF, Loureiro B, Hansen PJ - Sci Rep (2013)

Bottom Line: Treatment of bovine embryos with AMBMP at day 5 after insemination decreased development to the blastocyst stage at day 7 and reduced numbers of trophectoderm and inner cell mass cells.Examination of gene expression in day 6 morulae by microarray revealed expression of 16 WNT genes and other genes involved in WNT signaling; differences in relative expression were confirmed by PCR for 7 genes.In conclusion, the preimplantation embryo possesses a functional WNT signaling system and activation of the canonical pathway can inhibit embryonic development.

View Article: PubMed Central - PubMed

Affiliation: Department of Animal Sciences, D.H. Barron Reproductive and Perinatal Biology Program, and Genetics Institute, University of Florida, Gainesville, FL 32611-0910, USA.

ABSTRACT
Objectives were to evaluate the role of canonical WNT signaling in development of the preimplantation embryo. Signaling was activated with 2-Amino-4-(3,4-(methylenedioxy)benzylamino)-6-(3-methoxyphenyl)pyrimidine (AMBMP) and inhibited with Dickkopf-related protein 1 (DKK1). Treatment of bovine embryos with AMBMP at day 5 after insemination decreased development to the blastocyst stage at day 7 and reduced numbers of trophectoderm and inner cell mass cells. At high concentrations, AMBMP caused disorganization of the inner cell mass. DKK1 blocked actions of AMBMP but did not affect development in the absence of AMBMP. Examination of gene expression in day 6 morulae by microarray revealed expression of 16 WNT genes and other genes involved in WNT signaling; differences in relative expression were confirmed by PCR for 7 genes. In conclusion, the preimplantation embryo possesses a functional WNT signaling system and activation of the canonical pathway can inhibit embryonic development.

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Genes related to WNT signaling are expressed in day 6 morulae.(A) Agarose gel electrophoretogram of reverse transcriptase PCR products for CTNNBL1, DKK1, FZD7, LRP6, RYK, WNT11, SDHA and GAPDH. Note that another gene, WNT3, was not detectable. (B) Correlation between expression results for day 6 morulae obtained from microarray analysis and from real-time PCR. WNT3 was undetectable and the amounts of other mRNA were expressed as a fold-change relative to amount of WNT11.
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f4: Genes related to WNT signaling are expressed in day 6 morulae.(A) Agarose gel electrophoretogram of reverse transcriptase PCR products for CTNNBL1, DKK1, FZD7, LRP6, RYK, WNT11, SDHA and GAPDH. Note that another gene, WNT3, was not detectable. (B) Correlation between expression results for day 6 morulae obtained from microarray analysis and from real-time PCR. WNT3 was undetectable and the amounts of other mRNA were expressed as a fold-change relative to amount of WNT11.

Mentions: Analysis of a separate set of embryos at day 6 of development by qPCR confirmed results of the microarray. A total of 7 genes were analyzed and all but one (WNT3) were detectable by qPCR (Figure 4A). Moreover, the amount of mRNA, expressed as the fold-change relative to WNT11 (the least-abundant detectable gene) was highly correlated (r = 0.83, P = 0.02) with amount of mRNA detected by microarray analysis (Figure 4B).


Canonical WNT signaling regulates development of bovine embryos to the blastocyst stage.

Denicol AC, Dobbs KB, McLean KM, Carambula SF, Loureiro B, Hansen PJ - Sci Rep (2013)

Genes related to WNT signaling are expressed in day 6 morulae.(A) Agarose gel electrophoretogram of reverse transcriptase PCR products for CTNNBL1, DKK1, FZD7, LRP6, RYK, WNT11, SDHA and GAPDH. Note that another gene, WNT3, was not detectable. (B) Correlation between expression results for day 6 morulae obtained from microarray analysis and from real-time PCR. WNT3 was undetectable and the amounts of other mRNA were expressed as a fold-change relative to amount of WNT11.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3569626&req=5

f4: Genes related to WNT signaling are expressed in day 6 morulae.(A) Agarose gel electrophoretogram of reverse transcriptase PCR products for CTNNBL1, DKK1, FZD7, LRP6, RYK, WNT11, SDHA and GAPDH. Note that another gene, WNT3, was not detectable. (B) Correlation between expression results for day 6 morulae obtained from microarray analysis and from real-time PCR. WNT3 was undetectable and the amounts of other mRNA were expressed as a fold-change relative to amount of WNT11.
Mentions: Analysis of a separate set of embryos at day 6 of development by qPCR confirmed results of the microarray. A total of 7 genes were analyzed and all but one (WNT3) were detectable by qPCR (Figure 4A). Moreover, the amount of mRNA, expressed as the fold-change relative to WNT11 (the least-abundant detectable gene) was highly correlated (r = 0.83, P = 0.02) with amount of mRNA detected by microarray analysis (Figure 4B).

Bottom Line: Treatment of bovine embryos with AMBMP at day 5 after insemination decreased development to the blastocyst stage at day 7 and reduced numbers of trophectoderm and inner cell mass cells.Examination of gene expression in day 6 morulae by microarray revealed expression of 16 WNT genes and other genes involved in WNT signaling; differences in relative expression were confirmed by PCR for 7 genes.In conclusion, the preimplantation embryo possesses a functional WNT signaling system and activation of the canonical pathway can inhibit embryonic development.

View Article: PubMed Central - PubMed

Affiliation: Department of Animal Sciences, D.H. Barron Reproductive and Perinatal Biology Program, and Genetics Institute, University of Florida, Gainesville, FL 32611-0910, USA.

ABSTRACT
Objectives were to evaluate the role of canonical WNT signaling in development of the preimplantation embryo. Signaling was activated with 2-Amino-4-(3,4-(methylenedioxy)benzylamino)-6-(3-methoxyphenyl)pyrimidine (AMBMP) and inhibited with Dickkopf-related protein 1 (DKK1). Treatment of bovine embryos with AMBMP at day 5 after insemination decreased development to the blastocyst stage at day 7 and reduced numbers of trophectoderm and inner cell mass cells. At high concentrations, AMBMP caused disorganization of the inner cell mass. DKK1 blocked actions of AMBMP but did not affect development in the absence of AMBMP. Examination of gene expression in day 6 morulae by microarray revealed expression of 16 WNT genes and other genes involved in WNT signaling; differences in relative expression were confirmed by PCR for 7 genes. In conclusion, the preimplantation embryo possesses a functional WNT signaling system and activation of the canonical pathway can inhibit embryonic development.

Show MeSH
Related in: MedlinePlus