title: Mammary epithelial tubes elongate through MAPK-dependent coordination of cell migration.
abstract: Mammary branching morphogenesis is regulated by receptor tyrosine kinases (RTKs). We sought to determine how these RTK signals alter proliferation and migration to accomplish tube elongation in mouse. Both behaviors occur but it has been difficult to determine their relative contribution to elongation in vivo, as mammary adipocytes scatter light and limit the depth of optical imaging. Accordingly, we utilized 3D culture to study elongation in an experimentally accessible setting. We first used antibodies to localize RTK signals and discovered that phosphorylated ERK1/2 (pERK) was spatially enriched in cells near the front of elongating ducts, whereas phosphorylated AKT was ubiquitous. We next observed a gradient of cell migration speeds from rear to front of elongating ducts, with the front characterized by both high pERK and the fastest cells. Furthermore, cells within elongating ducts oriented both their protrusions and their migration in the direction of tube elongation. By contrast, cells within the organoid body were isotropically protrusive. We next tested the requirement for proliferation and migration. Early inhibition of proliferation blocked the creation of migratory cells, whereas late inhibition of proliferation did not block continued duct elongation. By contrast, pharmacological inhibition of either MEK or Rac1 signaling acutely blocked both cell migration and duct elongation. Finally, conditional induction of MEK activity was sufficient to induce collective cell migration and ductal elongation. Our data suggest a model for ductal elongation in which RTK-dependent proliferation creates motile cells with high pERK, the collective migration of which acutely requires both MEK and Rac1 signaling.
authors.affiliation: Departments of Cell Biology, Oncology, and Biomedical Engineering, Center for Cell Dynamics, Johns Hopkins University School of Medicine, 855 N. Wolfe Street, 452 Rangos Building, Baltimore, MD 21205, USA.
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Huebner
authors.affiliation: Departments of Cell Biology, Oncology, and Biomedical Engineering, Center for Cell Dynamics, Johns Hopkins University School of Medicine, 855 N. Wolfe Street, 452 Rangos Building, Baltimore, MD 21205, USA.
authors.forename: Neil M
authors.initials: NM
authors.lastname: Neumann
authors.affiliation: Departments of Cell Biology, Oncology, and Biomedical Engineering, Center for Cell Dynamics, Johns Hopkins University School of Medicine, 855 N. Wolfe Street, 452 Rangos Building, Baltimore, MD 21205, USA andrew.ewald@jhmi.edu.
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Ewald
mesh_terms: D000818:Animals; D016880:Anisotropy; D002465:Cell Movement; D049109:Cell Proliferation; D004847:Epithelial Cells / Q000166:cytology*; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism*; D005260:Female; D006801:Humans; D008321:Mammary Glands, Animal / Q000166:cytology*; D008822:Mice, Transgenic; D020929:Mitogen-Activated Protein Kinase Kinases / Q000378:metabolism; D009024:Morphogenesis; D009940:Organoids / Q000378:metabolism; D010766:Phosphorylation; D015398:Signal Transduction; D020830:rac1 GTP-Binding Protein / Q000378:metabolism
chemical_list: D048049:Extracellular Signal-Regulated MAP Kinases; D020929:Mitogen-Activated Protein Kinase Kinases; D020830:rac1 GTP-Binding Protein
keywords: Branching morphogenesis; Collective cell migration; Epithelial development; MAPK signaling; Mammary gland; Tubulogenesis
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