title: High-efficiency type II cell-enhanced green fluorescent protein expression facilitates cellular identification, tracking, and isolation.
abstract: We have developed a transgenic mouse expressing enhanced green fluorescent protein (EGFP) in virtually all type II (TII) alveolar epithelial cells. The CBG mouse (SPC-BAC-EGFP) contains a bacterial artificial chromosome modified to express EGFP within the mouse surfactant protein (SP)-C gene 3' untranslated region. EGFP mRNA expression is limited to the lung. EGFP fluorescence is both limited to and exhibited by all cells expressing pro-SP-C; fluorescence is uniform throughout all lobes of the lung and does not change as mice age. EGFP(+) cells also express SP-B but do not express podoplanin, a type I (TI) cell marker. CBG mice show no evidence of lung disease with aging. In 3 hours, TII cells can be isolated in >99% purity from CBG mice by FACS; the yield of 3.7 ± 0.6 × 10(6) cells represents approximately 25 to 60% of the TII cells in the lung. By FACS analysis, approximately 0.9% of TII cells are in mitosis in uninjured lungs; after bleomycin injury, 4.1% are in mitosis. Because EGFP fluorescence can be detected for >14 days in culture, at a time that SP-C mRNA expression is essentially nil, this line may be useful for tracking TII cells in culture and in vivo. When CBG mice are crossed to transgenic mice expressing rat podoplanin, TI and TII cells can be easily simultaneously identified and isolated. When bred to other strains of mice, EGFP expression can be used to identify TII cells without the need for immunostaining for SP-C. These mice should be useful in models of mouse pulmonary disease and in studies of TII cell biology, biochemistry, and genetics.
authors.affiliation: 1 Cardiovascular Research Institute and.
authors.forename: Jeff N
authors.initials: JN
authors.lastname: Vanderbilt
authors.affiliation: 1 Cardiovascular Research Institute and.
authors.forename: Robert F
authors.initials: RF
authors.lastname: Gonzalez
authors.affiliation: 1 Cardiovascular Research Institute and.
authors.forename: Lennell
authors.initials: L
authors.lastname: Allen
authors.affiliation: 1 Cardiovascular Research Institute and.
authors.forename: AnneMarie
authors.initials: A
authors.lastname: Gillespie
authors.affiliation: 1 Cardiovascular Research Institute and.
authors.forename: David
authors.initials: D
authors.lastname: Leaffer
authors.affiliation: 1 Cardiovascular Research Institute and.
authors.forename: Willow B
authors.initials: WB
authors.lastname: Dean
authors.affiliation: Departments of 2 Pediatrics and.
authors.forename: Cheryl
authors.initials: C
authors.lastname: Chapin
authors.affiliation: 1 Cardiovascular Research Institute and.|Departments of 2 Pediatrics and.|3 Medicine, University of California, San Francisco, San Francisco, California.
authors.forename: Leland G
authors.initials: LG
authors.lastname: Dobbs
mesh_terms: D000818:Animals; D000943:Antigens, Differentiation / Q000235:genetics / Q000378:metabolism; D002469:Cell Separation; D058948:Cell Tracking; D022202:Chromosomes, Artificial, Bacterial; D004195:Disease Models, Animal; D015870:Gene Expression; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D036341:Intercellular Signaling Peptides and Proteins; D008168:Lung / Q000166:cytology / Q000378:metabolism; D008171:Lung Diseases / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D010455:Peptides / Q000235:genetics / Q000378:metabolism; D037721:Pulmonary Surfactant-Associated Protein C; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D000098853:Podoplanin
chemical_list: D000943:Antigens, Differentiation; D049452:Green Fluorescent Proteins; D036341:Intercellular Signaling Peptides and Proteins; D008562:Membrane Glycoproteins; D010455:Peptides; D037721:Pulmonary Surfactant-Associated Protein C; D011993:Recombinant Fusion Proteins; C092025:Pdpn protein, rat; D000098853:Podoplanin; C489680:Sftpc protein, mouse; C485184:enhanced green fluorescent protein
keywords: alveolar epithelium; alveolar type II cells; bacterial artificial chromosome; enhanced green fluorescent protein; surfactant protein C