title: Screening for activators of the wingless type/Frizzled pathway by automated fluorescent microscopy.
abstract: Development of means to screen primary human cells rather than established cell lines is important in improving the predictive value of cellular assays in drug discovery. We describe a method of using automated fluorescent microscopy to detect activators of the wingless type/Frizzled (Wnt/Fzd) pathway in primary human preosteoblasts. This technique relies on detection of endogenous beta-catenin translocation to the nucleus as an indicator of pathway activation, requires only a limited number of primary cells, and is robust enough for automation and high-content, high-throughput screening. Identification of activators of the Wnt/Fzd pathway in human preosteoblasts may be useful in providing lead compounds for the treatment of osteoporosis.
authors.affiliation: Becton-Dickinson Technologies, Research Triangle Park, North Carolina, USA.
authors.forename: Kristen M
authors.initials: KM
authors.lastname: Borchert
authors.forename: Rachelle J
authors.initials: RJ
authors.lastname: Sells Galvin
authors.forename: Laura V
authors.initials: LV
authors.lastname: Hale
authors.forename: O Joseph
authors.initials: OJ
authors.lastname: Trask
authors.forename: Debra R
authors.initials: DR
authors.lastname: Nickischer
authors.forename: Keith A
authors.initials: KA
authors.lastname: Houck
mesh_terms: D021581:Active Transport, Cell Nucleus; D001331:Automation; D002454:Cell Differentiation; D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured; D017077:Culture Media, Conditioned / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D051157:Frizzled Receptors / Q000502:physiology*; D006801:Humans; D008856:Microscopy, Fluorescence / Q000379:methods*; D010006:Osteoblasts / Q000166:cytology* / Q000378:metabolism*; D010024:Osteoporosis / Q000628:therapy; D015534:Trans-Activators; D051153:Wnt Proteins / Q000502:physiology*; D051176:beta Catenin / Q000378:metabolism
chemical_list: D017077:Culture Media, Conditioned; D051157:Frizzled Receptors; D015534:Trans-Activators; D051153:Wnt Proteins; D051176:beta Catenin