Real time monitoring of BMP Smads transcriptional activity during mouse development

Bone morphogenetic protein (BMP) signaling is a key pathway in the patterning and development of organisms as diverse as fruit fly and humans. However, the determination of net BMP signaling, paramount to understanding organogenesis, is limited to the analysis of fixed material. We generated a transgenic mouse that reports the transcriptional response of BMP Smad activation by coupling a well established BMP response element (BRE), isolated from the Id1 promoter, to green fluorescent protein (BRE:gfp). We monitored BMP Smad transcriptional activity in fresh and fixed BRE:gfp embryos. GFP expression was observed where expected on the basis of known signaling sites, but also in specific cell populations in which BMP signaling had been implicated but not directly demonstrated. Deletion of Smad5 reduced GFP in vivo as expected. The BRE:gfp transgenic mice are a novel tool, which will facilitate the identification of specific BMP Smad responsive cell types and allow BMP Smad signaling to be monitored in real time, supporting studies in development and disease. genesis 46:335–346, 2008. © 2008 Wiley‐Liss, Inc.

[1]  W. Seeger,et al.  Temporal and spatial regulation of bone morphogenetic protein signaling in late lung development , 2007, Developmental dynamics : an official publication of the American Association of Anatomists.

[2]  Nathalie Perreault,et al.  Bone morphogenetic protein signaling is essential for terminal differentiation of the intestinal secretory cell lineage. , 2007, Gastroenterology.

[3]  M. V. Dinther,et al.  BMP-9 signals via ALK1 and inhibits bFGF-induced endothelial cell proliferation and VEGF-stimulated angiogenesis , 2007, Journal of Cell Science.

[4]  G. Robinson,et al.  BMP4 and PTHrP interact to stimulate ductal outgrowth during embryonic mammary development and to inhibit hair follicle induction , 2007, Development.

[5]  R. Kalluri,et al.  BMP‐7 functions as a novel hormone to facilitate liver regeneration , 2007, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[6]  R. Heuckeroth,et al.  BMP signaling regulates murine enteric nervous system precursor migration, neurite fasciculation, and patterning via altered Ncam1 polysialic acid addition. , 2006, Developmental biology.

[7]  J. Klingensmith,et al.  Morphogenesis of the trachea and esophagus: current players and new roles for noggin and Bmps. , 2006, Differentiation; research in biological diversity.

[8]  E. Bikoff,et al.  Dose-dependent Smad1, Smad5 and Smad8 signaling in the early mouse embryo. , 2006, Developmental biology.

[9]  Pascal Schneider,et al.  Generation of the primary hair follicle pattern. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[10]  R. Dietz,et al.  Requirement of Nuclear Factor-&kgr;B in Angiotensin II– and Isoproterenol-Induced Cardiac Hypertrophy In Vivo , 2005, Circulation.

[11]  D. Ovchinnikov,et al.  Bmpr1a and Bmpr1b have overlapping functions and are essential for chondrogenesis in vivo. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[12]  C. Tabin,et al.  Sox9 and Nkx2.5 determine the pyloric sphincter epithelium under the control of BMP signaling. , 2005, Developmental biology.

[13]  M. Goumans,et al.  Endoglin promotes endothelial cell proliferation and TGF‐β/ALK1 signal transduction , 2004, The EMBO journal.

[14]  C. Mummery,et al.  Spatio-temporal activation of Smad1 and Smad5 in vivo: monitoring transcriptional activity of Smad proteins , 2004, Journal of Cell Science.

[15]  D. Roberts,et al.  SOX9 specifies the pyloric sphincter epithelium through mesenchymal-epithelial signals , 2004, Development.

[16]  J. Epstein,et al.  Cardiac outflow tract defects in mice lacking ALK2 in neural crest cells , 2004, Development.

[17]  Philippe Soriano,et al.  In vivo convergence of BMP and MAPK signaling pathways: impact of differential Smad1 phosphorylation on development and homeostasis. , 2004, Genes & development.

[18]  R. Markwald,et al.  Bone morphogenetic protein-2 can mediate myocardial regulation of atrioventricular cushion mesenchymal cell formation in mice. , 2004, Developmental biology.

[19]  C. Chuong,et al.  Morpho-regulation of ectodermal organs: integument pathology and phenotypic variations in K14-Noggin engineered mice through modulation of bone morphogenic protein pathway. , 2004, The American journal of pathology.

[20]  M. Goumans,et al.  Controlling cell fate by bone morphogenetic protein receptors , 2003, Molecular and Cellular Endocrinology.

[21]  E. Fuchs,et al.  Defining BMP functions in the hair follicle by conditional ablation of BMP receptor IA , 2003, The Journal of cell biology.

[22]  TsugioSeki,et al.  Arterial Endothelium-Specific Activin Receptor-Like Kinase 1 Expression Suggests Its Role in Arterialization and Vascular Remodeling , 2003 .

[23]  J. Brissette,et al.  Noggin overexpression inhibits eyelid opening by altering epidermal apoptosis and differentiation , 2003, The EMBO journal.

[24]  P. Serup,et al.  Expression and misexpression of members of the FGF and TGFβ families of growth factors in the developing mouse pancreas , 2003, Developmental dynamics : an official publication of the American Association of Anatomists.

[25]  K. Lyons,et al.  BMP signaling is required for septation of the outflow tract of the mammalian heart , 2003, Development.

[26]  Y. Matsui,et al.  SMAD1 signaling is critical for initial commitment of germ cell lineage from mouse epiblast , 2002, Mechanisms of Development.

[27]  T. Johansen,et al.  Evaluation of Tet-on system to avoid transgene down-regulation in ex vivo gene transfer to the CNS , 2002, Gene Therapy.

[28]  R. Behringer,et al.  Endocardial cushion and myocardial defects after cardiac myocyte-specific conditional deletion of the bone morphogenetic protein receptor ALK3 , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[29]  P. ten Dijke,et al.  Identification and Functional Characterization of Distinct Critically Important Bone Morphogenetic Protein-specific Response Elements in the Id1 Promoter* , 2002, The Journal of Biological Chemistry.

[30]  E. Robertson,et al.  Mouse embryos lacking Smad1 signals display defects in extra-embryonic tissues and germ cell formation. , 2001, Development.

[31]  B. Hogan,et al.  Distinct mesodermal signals, including BMPs from the septum transversum mesenchyme, are required in combination for hepatogenesis from the endoderm. , 2001, Genes & development.

[32]  R. Kim,et al.  Bmp6 and Bmp7 are required for cushion formation and septation in the developing mouse heart. , 2001, Developmental biology.

[33]  Edward S. Kim,et al.  Immunohistochemical expression of smads 1–6 in the 15‐day gestation mouse embryo: signaling by BMPs and TGF‐βs † , 2001, Developmental dynamics : an official publication of the American Association of Anatomists.

[34]  K. Flanders,et al.  Characterization of the mouse Smad1 gene and its expression pattern in adult mouse tissues. , 2000, Gene.

[35]  J. Massagué,et al.  Transcriptional control by the TGF‐β/Smad signaling system , 2000 .

[36]  M. Goumans,et al.  Functional analysis of the TGFbeta receptor/Smad pathway through gene ablation in mice. , 2000, The International journal of developmental biology.

[37]  P. Donahoe,et al.  Activin receptor-like kinase 1 modulates transforming growth factor-beta 1 signaling in the regulation of angiogenesis. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[38]  K. Lyons,et al.  The type I BMP receptor BMPRIB is required for chondrogenesis in the mouse limb. , 2000, Development.

[39]  M. Wegner,et al.  Bone Morphogenetic Proteins Are Required In Vivo for the Generation of Sympathetic Neurons , 1999, Neuron.

[40]  B. Hogan,et al.  Bmp signaling regulates proximal-distal differentiation of endoderm in mouse lung development. , 1999, Development.

[41]  M. Matzuk,et al.  Smad5 knockout mice die at mid-gestation due to multiple embryonic and extraembryonic defects. , 1999, Development.

[42]  C. Ackerley,et al.  Astrocytoma cell interaction with elastin substrates: Implications for astrocytoma invasive potential , 1999, Glia.

[43]  B. Hogan,et al.  BMP4 is essential for lens induction in the mouse embryo. , 1998, Genes & development.

[44]  M. Mehler,et al.  Development of Bone Morphogenetic Protein Receptors in the Nervous System and Possible Roles in Regulating trkC Expression , 1998, The Journal of Neuroscience.

[45]  C. Mummery,et al.  Expression of ALK‐1, a type 1 serine/threonine kinase receptor, coincides with sites of vasculogenesis and angiogenesis in early mouse development , 1997, Developmental dynamics : an official publication of the American Association of Anatomists.

[46]  N. Jena,et al.  BMP7 null mutation in mice: developmental defects in skeleton, kidney, and eye. , 1997, Experimental cell research.

[47]  A. Bradley,et al.  Mice deficient for BMP2 are nonviable and have defects in amnion/chorion and cardiac development. , 1996, Development.

[48]  H. Rohrer,et al.  Involvement of bone morphogenetic protein-4 and bone morphogenetic protein-7 in the differentiation of the adrenergic phenotype in developing sympathetic neurons. , 1996, Development.

[49]  A. Bradley,et al.  BMP-7 is an inducer of nephrogenesis, and is also required for eye development and skeletal patterning. , 1995, Genes & development.

[50]  K. Lyons,et al.  A requirement for bone morphogenetic protein-7 during development of the mammalian kidney and eye. , 1995, Genes & development.

[51]  A. McMahon,et al.  Hedgehog and Bmp genes are coexpressed at many diverse sites of cell-cell interaction in the mouse embryo. , 1995, Developmental biology.

[52]  K. Miyazono,et al.  Distinct spatial and temporal expression patterns of two type I receptors for bone morphogenetic proteins during mouse embryogenesis. , 1995, Endocrinology.

[53]  B. Hogan,et al.  Colocalization of BMP 7 and BMP 2 RNAs suggests that these factors cooperatively mediate tissue interactions during murine development , 1995, Mechanisms of Development.

[54]  B. Hogan,et al.  Involvement of Bone Morphogenetic Protein-4 (BMP-4) and Vgr-1 in morphogenesis and neurogenesis in the mouse. , 1991, Development.

[55]  B. Hogan,et al.  Organogenesis and pattern formation in the mouse: RNA distribution patterns suggest a role for bone morphogenetic protein-2A (BMP-2A). , 1990, Development.

[56]  V. Rosen,et al.  Novel regulators of bone formation: molecular clones and activities. , 1988, Science.

[57]  M. Matzuk,et al.  Genetic analysis of the mammalian transforming growth factor-beta superfamily. , 2002, Endocrine reviews.

[58]  K. Miyazono,et al.  TGF-beta signalling from cell membrane to nucleus through SMAD proteins. , 1997, Nature.

[59]  B. Hogan.  Bmps: multifunctional regulators of mammalian embryonic development. , 1996, Harvey lectures.