title: Image-based screening identifies novel roles for IkappaB kinase and glycogen synthase kinase 3 in axonal degeneration.
abstract: Axon degeneration is an active, evolutionarily conserved self-destruction program by which compromised axons fragment in response to varied insults. Unlike programmed cell death, axon degeneration is poorly understood. We have combined robotic liquid handling with automated microscopy and image analysis to create a robust screening platform to measure axon degeneration in mammalian primary neuronal cultures. Using this assay, we performed an unbiased screen of 480 bioactive compounds, identifying 11 that reproducibly delay fragmentation of severed axons in vitro, including two inhibitors of glycogen synthase kinase 3 and two inhibitors of IκB kinase. Knockdown of each of these targets by shRNA lentivirus also delays axon degeneration in vitro, further supporting their role in the axon degeneration program.
authors.affiliation: Department of Genetics, Washington University Medical School, St Louis, Missouri 63110, USA.
authors.forename: Josiah
authors.initials: J
authors.lastname: Gerdts
authors.forename: Yo
authors.initials: Y
authors.lastname: Sasaki
authors.forename: Bhupinder
authors.initials: B
authors.lastname: Vohra
authors.forename: Jayne
authors.initials: J
authors.lastname: Marasa
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Milbrandt
mesh_terms: D000818:Animals; D001369:Axons / Q000201:enzymology*; D002478:Cells, Cultured; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D038362:Glycogen Synthase Kinase 3 / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D051550:I-kappa B Kinase / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008856:Microscopy, Fluorescence / Q000379:methods; D019636:Neurodegenerative Diseases / Q000188:drug therapy / Q000201:enzymology*
chemical_list: D004791:Enzyme Inhibitors; D051550:I-kappa B Kinase; D038362:Glycogen Synthase Kinase 3