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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 I?B 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.

SUBMITTER: Gerdts J

PROVIDER: S-EPMC3151046 | biostudies-literature | 2011 Aug

REPOSITORIES: biostudies-literature

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Image-based screening identifies novel roles for IkappaB kinase and glycogen synthase kinase 3 in axonal degeneration.

Gerdts Josiah J Sasaki Yo Y Vohra Bhupinder B Marasa Jayne J Milbrandt Jeffrey J

The Journal of biological chemistry 20110617 32

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, identifyin ...[more]

PMID: 21685387

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Image-based screening identifies novel roles for IkappaB kinase and glycogen synthase kinase 3 in axonal degeneration.

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Image-based screening identifies novel roles for IkappaB kinase and glycogen synthase kinase 3 in axonal degeneration.

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