title: A Cre-inducible fluorescent reporter for observing apical membrane dynamics.
abstract: The ability to image living tissues with fluorescent proteins has revolutionized the fields of cell and developmental biology. Fusions between fluorescent proteins and various polypeptides are allowing scientists to image tissues with sub-cellular resolution. Here, we describe the generation and activity of a genetically engineered mouse line expressing a fusion between the green fluorescent protein (GFP) and the apically localized protein Crumbs3 (Crb3). This reporter drives Cre-inducible expression of Crb3-GFP under control of the EF1a regulatory domains. The fusion protein is broadly expressed in embryonic and adult tissues and shows apical restriction in the majority of epithelial cell types. It displays a variably penetrant gain of function activity in the neural tube. However, in several cell types, over-expression of Crb3 does not appear to have any effect on normal development or maintenance. Detailed analysis of kidneys expressing this reporter indicates normal morphology and function highlighting the utility for live imaging. Thus, the EF1a(Crb3-GFP) mouse line will be of broad use for studying membrane and/or tissue dynamics in living tissues.
authors: Department of Internal Medicine (Nephrology), UT Southwestern Medical Center, Dallas, Texas; Department of Molecular Biology, UT Southwestern Medical Center, Dallas, Texas.
authors: Xinchao
authors: X
authors: Pan
authors: Ulrike
authors: U
authors: Schnell
authors: Courtney M
authors: CM
authors: Karner
authors: Erin V
authors: EV
authors: Small
authors: Thomas J
authors: TJ
authors: Carroll
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000378:metabolism*; D002478:Cells, Cultured; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism*; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism*; D005260:Female; D018507:Gene Expression Regulation, Developmental; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D007091:Image Processing, Computer-Assisted; D019426:Integrases / Q000378:metabolism*; D007668:Kidney / Q000166:cytology / Q000378:metabolism; D008562:Membrane Glycoproteins; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008822:Mice, Transgenic; D018613:Microscopy, Confocal; D009924:Organ Culture Techniques; D020648:Peptide Elongation Factor 1 / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics
chemical_list: C530695:Crb3 protein, mouse; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D020648:Peptide Elongation Factor 1; D049452:Green Fluorescent Proteins; C045073:Cre recombinase; D019426:Integrases
keywords: apical membrane; live imaging; membrane dynamics; morphogenesis