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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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Related in: MedlinePlus

Co-treatment of bovine embryos at day 5 after insemination with 100 ng/ml Dickkopf-related protein 1 (DKK1) reduces the inhibitory effect of a WNT agonist (AMBMP) on development to the blastocyst stage.Data are least-squares means ± SEM of results from 5 replicates. The percent of oocytes becoming blastocysts at day 7 was affected by agonist (P = 0.02) and the agonist by DKK1 interaction (P = 0.01).
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f2: Co-treatment of bovine embryos at day 5 after insemination with 100 ng/ml Dickkopf-related protein 1 (DKK1) reduces the inhibitory effect of a WNT agonist (AMBMP) on development to the blastocyst stage.Data are least-squares means ± SEM of results from 5 replicates. The percent of oocytes becoming blastocysts at day 7 was affected by agonist (P = 0.02) and the agonist by DKK1 interaction (P = 0.01).

Mentions: A third experiment was designed to test whether AMBMP blocked development by acting through the WNT signaling pathway. The approach was to determine whether DKK1, which prevents binding of the co-receptor LRP 5/6 to the agonist-receptor complex, blocks effects of AMBMP on development. As shown in Figure 2, AMBMP decreased (P = 0.02) the proportion of embryos that developed to the blastocyst stage at day 7. Moreover, blastocyst development was affected by the interaction between AMBMP and DKK1 at day 7 (P = 0.01). This interaction occurred as a result of blockage of the negative effect of AMBMP on blastocyst development by DKK1 (Figure 2); combination of AMBMP and DKK1 resulted in 22.2 ± 2.1% of embryos developing to blastocysts at day 7 compared to 15.9 ± 2.1% for AMBMP alone. In the absence of AMBMP, however, DKK1 tended to reduce blastocyst development at day 7 (29.9 versus 21.1 ± 2.1% for control and 100 ng/ml of DKK1, respectively).


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)

Co-treatment of bovine embryos at day 5 after insemination with 100 ng/ml Dickkopf-related protein 1 (DKK1) reduces the inhibitory effect of a WNT agonist (AMBMP) on development to the blastocyst stage.Data are least-squares means ± SEM of results from 5 replicates. The percent of oocytes becoming blastocysts at day 7 was affected by agonist (P = 0.02) and the agonist by DKK1 interaction (P = 0.01).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f2: Co-treatment of bovine embryos at day 5 after insemination with 100 ng/ml Dickkopf-related protein 1 (DKK1) reduces the inhibitory effect of a WNT agonist (AMBMP) on development to the blastocyst stage.Data are least-squares means ± SEM of results from 5 replicates. The percent of oocytes becoming blastocysts at day 7 was affected by agonist (P = 0.02) and the agonist by DKK1 interaction (P = 0.01).
Mentions: A third experiment was designed to test whether AMBMP blocked development by acting through the WNT signaling pathway. The approach was to determine whether DKK1, which prevents binding of the co-receptor LRP 5/6 to the agonist-receptor complex, blocks effects of AMBMP on development. As shown in Figure 2, AMBMP decreased (P = 0.02) the proportion of embryos that developed to the blastocyst stage at day 7. Moreover, blastocyst development was affected by the interaction between AMBMP and DKK1 at day 7 (P = 0.01). This interaction occurred as a result of blockage of the negative effect of AMBMP on blastocyst development by DKK1 (Figure 2); combination of AMBMP and DKK1 resulted in 22.2 ± 2.1% of embryos developing to blastocysts at day 7 compared to 15.9 ± 2.1% for AMBMP alone. In the absence of AMBMP, however, DKK1 tended to reduce blastocyst development at day 7 (29.9 versus 21.1 ± 2.1% for control and 100 ng/ml of DKK1, respectively).

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