A microfluidic culture platform for CNS axonal injury, regeneration and transport.
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.
Taylor
Anne M
AM
Department of Biomedical Engineering, University of California, Irvine, 204 Rockwell Engineering, Irvine, California 92697, USA.
Blurton-Jones
Mathew
M
Rhee
Seog Woo
SW
Cribbs
David H
DH
Cotman
Carl W
CW
Jeon
Noo Li
NL
D000818:Animals; D001369:Axons / Q000502:physiology* / Q000648:ultrastructure*; D019771:Axotomy / Q000379:methods*; D001693:Biological Transport, Active; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods; D002478:Cells, Cultured; D004867:Equipment Design; D019544:Equipment Failure Analysis; D017022:Flow Injection Analysis / Q000295:instrumentation* / Q000379:methods; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D009416:Nerve Regeneration / Q000502:physiology*; D059348:Peripheral Nerve Injuries; D010525:Peripheral Nerves / Q000473:pathology / Q000502:physiology* / Q000601:surgery; D051381:Rats