A Sprouty4 reporter to monitor FGF/ERK signaling activity in ESCs and mice Journal Article
| Authors: | Morgani, S. M.; Saiz, N.; Garg, V.; Raina, D.; Simon, C. S.; Kang, M.; Arias, A. M.; Nichols, J.; Schröter, C.; Hadjantonakis, A. K. |
| Article Title: | A Sprouty4 reporter to monitor FGF/ERK signaling activity in ESCs and mice |
| Abstract: | The FGF/ERK signaling pathway is highly conserved throughout evolution and plays fundamental roles during embryonic development and in adult organisms. While a plethora of expression data exists for ligands, receptors and pathway regulators, we know little about the spatial organization or dynamics of signaling in individual cells within populations. To this end we developed a transcriptional readout of FGF/ERK activity by targeting a histone H2B-linked Venus fluorophore to the endogenous locus of Spry4, an early pathway target, and generated Spry4H2B-Venus embryonic stem cells (ESCs) and a derivative mouse line. The Spry4H2B-Venus reporter was heterogeneously expressed within ESC cultures and responded to FGF/ERK signaling manipulation. In vivo, the Spry4H2B-Venus reporter recapitulated the expression pattern of Spry4 and localized to sites of known FGF/ERK activity including the inner cell mass of the pre-implantation embryo and the limb buds, somites and isthmus of the post-implantation embryo. Additionally, we observed highly localized reporter expression within adult organs. Genetic and chemical disruption of FGF/ERK signaling, in vivo in pre- and post-implantation embryos, abrogated Venus expression establishing the reporter as an accurate signaling readout. This tool will provide new insights into the dynamics of the FGF/ERK signaling pathway during mammalian development. © 2018 |
| Keywords: | signal transduction; mitogen activated protein kinase; adult; controlled study; genetics; nonhuman; animal cell; mouse; animal; cytology; metabolism; animals; mice; gene; cell survival; gene targeting; gene amplification; gene expression; map kinase signaling system; embryo; embryonic stem cell; nerve tissue proteins; gene locus; genotype; embryo development; embryology; in vivo study; physiology; cell lineage; reporter gene; extracellular signal-regulated map kinases; embryo, mammalian; cell count; histone h2b; nerve protein; fibroblast growth factor; fibroblast growth factors; embryonic development; gene location; cell tracking; live imaging; mouse embryo; organogenesis; nidation; quantitative image analysis; limb bud; procedures; fluorescent reporter; mapk signaling; somite; mammalian embryo; escs; mouse embryonic stem cells; male; female; priority journal; article; mouse embryonic stem cell; fgf signaling; fgf/erk; sprouty4; spry4 protein, mouse; sprouty4 gene |
| Journal Title: | Developmental Biology |
| Volume: | 441 |
| Issue: | 1 |
| ISSN: | 0012-1606 |
| Publisher: | Elsevier Inc. |
| Date Published: | 2018-09-01 |
| Start Page: | 104 |
| End Page: | 126 |
| Language: | English |
| DOI: | 10.1016/j.ydbio.2018.06.017 |
| PUBMED: | 29964027 |
| PROVIDER: | scopus |
| PMCID: | PMC6455974 |
| DOI/URL: | |
| Notes: | Article -- Export Date: 4 September 2018 -- Source: Scopus |
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10Morgani -
8Simon
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