A microfluidic culture platform for CNS axonal injury, regeneration and transport

Anne Taylor

Mathew Blurton-Jones

Seog Rhee

David Cribbs

Carl Cotman

Noo Jeon

^{Department of Biomedical Engineering, University of California, Irvine, 204 Rockwell Engineering, Irvine, California 92697, USA.}

^{Institute for Brain Aging and Dementia, University of California, Irvine, 1113 Gillespie NRF, Irvine, California 92697, USA.}

^{Department of Neurology, University of California, Irvine, 1207 Gillespie NRF, Irvine, California 92697, USA.}

^{These authors contributed equally to this work.}

Correspondence should be addressed to N.L.J. (ude.icu@noejn).

Abstract

Investigation of axonal biology in the central nervous system (CNS) is hindered by a lack of an appropriate in vitro method to probe axons independently from cell bodies. Here we describe a microfluidic culture platform that polarizes the growth of CNS axons into a fluidically isolated environment without the use of targeting neurotrophins. In addition to its compatibility with live cell imaging, the platform can be used to (i) isolate CNS axons without somata or dendrites, facilitating biochemical analyses of pure axonal fractions and (ii) localize physical and chemical treatments to axons or somata. We report the first evidence that presynaptic (Syp) but not postsynaptic (Camk2a) mRNA is localized to developing rat cortical and hippocampal axons. The platform also serves as a straightforward, reproducible method to model CNS axonal injury and regeneration. The results presented here demonstrate several experimental paradigms using the microfluidic platform, which can greatly facilitate future studies in axonal biology.

Abstract

Footnotes

Note: Supplementary information is available on the Nature Methods website.

COMPETING INTERESTS STATEMENT

The authors declare competing financial interests (see the Nature Methods website for details).

Footnotes

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