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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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Effect of administration of different concentrations of Dickkopf related protein 1 (DKK1) at day 5 after insemination on blastocyst development at day 7.Data are least-squares means ± SEM of results from 7 replicates. No difference in the percent of ooyctes that became blastocysts at day 7 after insemination was observed following treatment with DKK1 at concentrations between 50 and 400 ng/ml (P = 0.47).
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f3: Effect of administration of different concentrations of Dickkopf related protein 1 (DKK1) at day 5 after insemination on blastocyst development at day 7.Data are least-squares means ± SEM of results from 7 replicates. No difference in the percent of ooyctes that became blastocysts at day 7 after insemination was observed following treatment with DKK1 at concentrations between 50 and 400 ng/ml (P = 0.47).

Mentions: Because there was a tendency of DKK1 to decrease development, an additional experiment was performed to determine whether DKK1 itself has deleterious effects on embryo competence to develop to the blastocyst stage. Embryos were cultured with concentrations of DKK1 ranging from 0 to 400 ng/ml, beginning at day 5 after insemination. As shown in Figure 3, addition of DKK1 at concentrations of 50, 100, 200 and 400 ng/ml did not change (P = 0.47) the proportion of embryos reaching the blastocyst stage at day 7 of development as compared to embryos cultured without DKK1.


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)

Effect of administration of different concentrations of Dickkopf related protein 1 (DKK1) at day 5 after insemination on blastocyst development at day 7.Data are least-squares means ± SEM of results from 7 replicates. No difference in the percent of ooyctes that became blastocysts at day 7 after insemination was observed following treatment with DKK1 at concentrations between 50 and 400 ng/ml (P = 0.47).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f3: Effect of administration of different concentrations of Dickkopf related protein 1 (DKK1) at day 5 after insemination on blastocyst development at day 7.Data are least-squares means ± SEM of results from 7 replicates. No difference in the percent of ooyctes that became blastocysts at day 7 after insemination was observed following treatment with DKK1 at concentrations between 50 and 400 ng/ml (P = 0.47).
Mentions: Because there was a tendency of DKK1 to decrease development, an additional experiment was performed to determine whether DKK1 itself has deleterious effects on embryo competence to develop to the blastocyst stage. Embryos were cultured with concentrations of DKK1 ranging from 0 to 400 ng/ml, beginning at day 5 after insemination. As shown in Figure 3, addition of DKK1 at concentrations of 50, 100, 200 and 400 ng/ml did not change (P = 0.47) the proportion of embryos reaching the blastocyst stage at day 7 of development as compared to embryos cultured without DKK1.

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