Limits...
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.

Show MeSH

Related in: MedlinePlus

Representative images of labeling of inner cell mass and trophectoderm cells.Blastocysts were harvested at day 7 and subjected to differential immunofluorescence. Blue: Hoescht 33342 staining (all nuclei); green: CDX2+ cells (labeled with mouse anti-CDX2 and FITC-conjugated anti mouse IgG). Trophectoderm cells were labeled with Hoescht and anti CDX2 while inner cell mass cells were labeled with Hoescht only.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3569626&req=5

f1: Representative images of labeling of inner cell mass and trophectoderm cells.Blastocysts were harvested at day 7 and subjected to differential immunofluorescence. Blue: Hoescht 33342 staining (all nuclei); green: CDX2+ cells (labeled with mouse anti-CDX2 and FITC-conjugated anti mouse IgG). Trophectoderm cells were labeled with Hoescht and anti CDX2 while inner cell mass cells were labeled with Hoescht only.

Mentions: AMBMP reduced significantly total cell number at concentrations of 0.7 μM and higher (Table 1). Similarly, AMBMP reduced significantly numbers of trophectoderm (TE) cells at 1.4 μM and higher and numbers of ICM cells at 0.7 and 2.8 μM (Table 1). The ratio of ICM/TE cells was not significantly affected (P = 0.33) by AMBMP but the ratios were numerically highest at the two highest concentrations of AMBMP. At the highest concentrations of AMBMP, the ICM appeared disorganized with several cells not labeled with CDX2 (i.e. ICM) being located outside the cluster of cells characterized by the ICM (Figure 1).


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)

Representative images of labeling of inner cell mass and trophectoderm cells.Blastocysts were harvested at day 7 and subjected to differential immunofluorescence. Blue: Hoescht 33342 staining (all nuclei); green: CDX2+ cells (labeled with mouse anti-CDX2 and FITC-conjugated anti mouse IgG). Trophectoderm cells were labeled with Hoescht and anti CDX2 while inner cell mass cells were labeled with Hoescht only.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f1: Representative images of labeling of inner cell mass and trophectoderm cells.Blastocysts were harvested at day 7 and subjected to differential immunofluorescence. Blue: Hoescht 33342 staining (all nuclei); green: CDX2+ cells (labeled with mouse anti-CDX2 and FITC-conjugated anti mouse IgG). Trophectoderm cells were labeled with Hoescht and anti CDX2 while inner cell mass cells were labeled with Hoescht only.
Mentions: AMBMP reduced significantly total cell number at concentrations of 0.7 μM and higher (Table 1). Similarly, AMBMP reduced significantly numbers of trophectoderm (TE) cells at 1.4 μM and higher and numbers of ICM cells at 0.7 and 2.8 μM (Table 1). The ratio of ICM/TE cells was not significantly affected (P = 0.33) by AMBMP but the ratios were numerically highest at the two highest concentrations of AMBMP. At the highest concentrations of AMBMP, the ICM appeared disorganized with several cells not labeled with CDX2 (i.e. ICM) being located outside the cluster of cells characterized by the ICM (Figure 1).

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