Screening for activators of the wingless type/Frizzled pathway by automated fluorescent microscopy.
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.
Borchert
Kristen M
KM
Becton-Dickinson Technologies, Research Triangle Park, North Carolina, USA.
Sells Galvin
Rachelle J
RJ
Hale
Laura V
LV
Trask
O Joseph
OJ
Nickischer
Debra R
DR
Houck
Keith A
KA
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
D017077:Culture Media, Conditioned; D051157:Frizzled Receptors; D015534:Trans-Activators; D051153:Wnt Proteins; D051176:beta Catenin