Inactivation of nuclear Wnt- (cid:2) -catenin signaling limits blastocyst competency for implantation

l of plasma obtained by retroorbital phlebotomy from pregnant females receiving empty or Dkk1 ADV using anti-His probe antibody (Santa Cruz Biotechnology) as previously described (Kuhnert et al., 2004). To further explore the consequence of Wnt inhibition on blastocyst implantation, we applied small-molecule inhibitors of the (cid:2) -catenin-Tcf complex, PKF115-584 and CGP049090 (Novartis) (Lepourcelet The activation of the blastocyst, a process by which it gains competency to attach with the receptive uterus, is a prerequisite for successful implantation. However, the molecular basis of blastocyst activation remains largely unexplored. Combining molecular, pharmacological and physiological approaches, we show here that silencing of Wnt- (cid:2) -catenin signaling in mice does not adversely affect the development of preimplantation embryos to blastocysts and uterine preparation for receptivity, but, remarkably, blocks blastocyst competency to implantation. Using the physiologically relevant delayed implantation model and trophoblast stem cells in culture, we further demonstrate that a coordinated activation of canonical Wnt- (cid:2) -catenin signaling with attenuation of the non-canonical Wnt-RhoA signaling pathway ensures blastocyst competency to implantation. These findings constitute novel evidence that Wnt signaling is at least one pathway that determines blastocyst competency for implantation.

[1]  M. Zernicka-Goetz,et al.  Dishevelled proteins regulate cell adhesion in mouse blastocyst and serve to monitor changes in Wnt signaling. , 2007, Developmental biology.

[2]  Michael Kyba,et al.  Canonical Wnt signaling is required for development of embryonic stem cell-derived mesoderm , 2006, Development.

[3]  R. Nusse,et al.  Wnt Signaling: Multiple Pathways, Multiple Receptors, and Multiple Transcription Factors* , 2006, Journal of Biological Chemistry.

[4]  K. Jones,et al.  Wnt signaling: is the party in the nucleus? , 2006, Genes & development.

[5]  Jeffery R Barrow,et al.  Wnt/PCP signaling: a veritable polar star in establishing patterns of polarity in embryonic tissues. , 2006, Seminars in cell & developmental biology.

[6]  Sudhansu K. Dey,et al.  Roadmap to embryo implantation: clues from mouse models , 2006, Nature Reviews Genetics.

[7]  R. Jaenisch,et al.  Generation of nuclear transfer-derived pluripotent ES cells from cloned Cdx2-deficient blastocysts , 2006, Nature.

[8]  Janet Rossant,et al.  Interaction between Oct3/4 and Cdx2 Determines Trophectoderm Differentiation , 2005, Cell.

[9]  Bulent Ataman,et al.  Wingless Signaling at Synapses Is Through Cleavage and Nuclear Import of Receptor DFrizzled2 , 2005, Science.

[10]  E. Willems,et al.  Expression of all Wnt genes and their secreted antagonists during mouse blastocyst and postimplantation development , 2005, Developmental dynamics : an official publication of the American Association of Anatomists.

[11]  G. Bae,et al.  Nuclear localization is required for Dishevelled function in Wnt/β-catenin signaling , 2005, Journal of biology.

[12]  D. Dufort,et al.  Uterine Wnt/beta-catenin signaling is required for implantation. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[13]  D. Solter,et al.  Stabilization of β-catenin in the mouse zygote leads to premature epithelial-mesenchymal transition in the epiblast , 2004, Development.

[14]  G. Martin,et al.  Active Rho is localized to podosomes induced by oncogenic Src and is required for their assembly and function , 2004, The Journal of cell biology.

[15]  M. Ko,et al.  Global gene expression analysis identifies molecular pathways distinguishing blastocyst dormancy and activation. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[16]  A. Sharov,et al.  Dynamics of global gene expression changes during mouse preimplantation development. , 2004, Developmental cell.

[17]  A. Evsikov,et al.  Maternal beta-catenin and E-cadherin in mouse development. , 2004, Development.

[18]  Frank Petersen,et al.  Small-molecule antagonists of the oncogenic Tcf/beta-catenin protein complex. , 2004, Cancer cell.

[19]  Ronald W Davis,et al.  A genome-wide study of gene activity reveals developmental signaling pathways in the preimplantation mouse embryo. , 2004, Developmental cell.

[20]  Pauline Chu,et al.  Essential requirement for Wnt signaling in proliferation of adult small intestine and colon revealed by adenoviral expression of Dickkopf-1 , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[21]  R. Roberts,et al.  Differential G protein-coupled cannabinoid receptor signaling by anandamide directs blastocyst activation for implantation , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[22]  M. Murakami,et al.  The Homeoprotein Nanog Is Required for Maintenance of Pluripotency in Mouse Epiblast and ES Cells , 2003, Cell.

[23]  J. Nichols,et al.  Functional Expression Cloning of Nanog, a Pluripotency Sustaining Factor in Embryonic Stem Cells , 2003, Cell.

[24]  Christof Niehrs,et al.  Kremen2 modulates Dickkopf2 activity during Wnt/LRP6 signaling. , 2003, Gene.

[25]  Randall T Moon,et al.  A second canon. Functions and mechanisms of beta-catenin-independent Wnt signaling. , 2003, Developmental cell.

[26]  M. Overduin,et al.  The DIX domain targets dishevelled to actin stress fibres and vesicular membranes , 2002, Nature.

[27]  E. Sahai,et al.  ROCK and Dia have opposing effects on adherens junctions downstream of Rho , 2002, Nature Cell Biology.

[28]  Christof Niehrs,et al.  Kremen proteins are Dickkopf receptors that regulate Wnt/beta-catenin signalling. , 2002, Nature.

[29]  Stuart A. Aaronson,et al.  Novel mechanism of Wnt signalling inhibition mediated by Dickkopf-1 interaction with LRP6/Arrow , 2001, Nature Cell Biology.

[30]  Michael Kühl,et al.  Head inducer Dickkopf-1 is a ligand for Wnt coreceptor LRP6 , 2001, Current Biology.

[31]  Yan Li,et al.  LDL-receptor-related protein 6 is a receptor for Dickkopf proteins , 2001, Nature.

[32]  S. Kliewer,et al.  A selective peroxisome proliferator-activated receptor δ agonist promotes reverse cholesterol transport , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[33]  B. Hogan,et al.  Cellular and molecular responses of the uterus to embryo implantation can be elicited by locally applied growth factors. , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[34]  W. Nelson,et al.  Colocalization and Redistribution of Dishevelled and Actin during WNT-Induced Mesenchymal Morphogenesis , 2000, The Journal of cell biology.

[35]  Carmen Birchmeier,et al.  Requirement for beta-catenin in anterior-posterior axis formation in mice. , 2000 .

[36]  R. Maas,et al.  Hoxa-10 regulates uterine stromal cell responsiveness to progesterone during implantation and decidualization in the mouse. , 1999, Molecular endocrinology.

[37]  K. Kinzler,et al.  PPARdelta is an APC-regulated target of nonsteroidal anti-inflammatory drugs. , 1999, Cell.

[38]  J. Rossant,et al.  Promotion of trophoblast stem cell proliferation by FGF4. , 1998, Science.

[39]  S. Dey,et al.  Coordination of differential effects of primary estrogen and catecholestrogen on two distinct targets mediates embryo implantation in the mouse. , 1998, Endocrinology.

[40]  A. Sparks,et al.  Identification of c-MYC as a target of the APC pathway. , 1998, Science.

[41]  C. Niehrs,et al.  Dickkopf-1 is a member of a new family of secreted proteins and functions in head induction , 1998, Nature.

[42]  A. Hall,et al.  The Small GTPases Rho and Rac Are Required for the Establishment of Cadherin-dependent Cell–Cell Contacts , 1997, The Journal of cell biology.

[43]  J. Nathans,et al.  A family of secreted proteins contains homology to the cysteine-rich ligand-binding domain of frizzled receptors. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[44]  L. Larue,et al.  Lack of beta-catenin affects mouse development at gastrulation. , 1995, Development.

[45]  S. Dey,et al.  Amphiregulin is an implantation-specific and progesterone-regulated gene in the mouse uterus. , 1995, Molecular endocrinology.

[46]  M. Klagsbrun,et al.  Heparin-binding EGF-like growth factor gene is induced in the mouse uterus temporally by the blastocyst solely at the site of its apposition: a possible ligand for interaction with blastocyst EGF-receptor in implantation. , 1994, Development.

[47]  S. Dey,et al.  Blastocyst's state of activity determines the "window" of implantation in the receptive mouse uterus. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[48]  S. Dey,et al.  Antisense c-myc effects on preimplantation mouse embryo development. , 1992, Proceedings of the National Academy of Sciences of the United States of America.