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MBC in Press, published online ahead of print November 8, 2006
Mol. Biol. Cell 10.1091/mbc.E06-08-0666

A more recent version of this article appeared on January 1, 2007
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Submitted on August 2, 2006
Revised on October 2, 2006
Accepted on October 27, 2006

High-Content Microscopy Identifies New Neurite Outgrowth Regulators

Vibor Laketa,* Jeremy C. Simpson,* Stephanie Bechtel,{dagger} Stefan Wiemann,{dagger} and Rainer Pepperkok*

*Cell Biology and Biophysics Unit, EMBL-Heidelberg, 69117 Heidelberg, Germany; {dagger}Division of Molecular Genome Analysis, German Cancer Research Center, 69120 Heidelberg, Germany

Monitoring Editor: Anne Ridley

Neurons, with their long axons and elaborate dendritic arbour, establish the complex circuitry that is essential for the proper functioning of the nervous system. While a catalogue of structural, molecular and functional differences between axons and dendrites is accumulating, the mechanisms involved in early events of neuronal differentiation, such as neurite initiation and elongation, are less well understood, mainly because the key molecules involved remain elusive. Here we describe the establishment and application of a microscopy-based approach designed to identify novel proteins involved in neurite initiation and/or elongation. We identified 21 proteins that affected neurite outgrowth when ectopically expressed in cells. Complementary time-lapse microscopy allowed us to discriminate between early and late effector proteins. Localization experiments with GFP-tagged proteins in fixed and living cells revealed a further 14 proteins that associated with neurite tips either early or late during neurite outgrowth. Coexpression experiments of the new effector proteins provide a first glimpse on a possible functional relationship of these proteins during neurite outgrowth. Altogether, we demonstrate the potential of the systematic microscope-based screening approaches described here to tackle the complex biological process of neurite outgrowth regulation.

Address correspondence to: Vibor Laketa (laketa{at}embl.de)






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