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Microfluidic culture platform for studying neuronal response to mild to very mild axonal stretch injury
journal contribution
posted on 2014-01-01, 00:00 authored by Y C Yap, T C Dickson, A E King, M C Breadmore, Rosanne GuijtRosanne GuijtA new model for studying localised axonal stretch injury is presented, using a microfluidic device to selectively culture axons on a thin, flexible poly (dimethylsiloxane) membrane which can be deflected upward to stretch the axons. A very mild (0.5% strain) or mild stretch injury (5% strain) was applied to primary cortical neurons after 7 days growth in vitro. The extent of distal degeneration was quantified using the degenerative index (DI, the ratio of fragmented axon area to total axon area) of axons fixed at 24 h and 72 h post injury (PI), and immunolabelled for the axon specific, microtubule associated protein-tau. At 24 h PI following very mild injuries (0.5%), the majority of the axons remained intact and healthy with no significant difference in DI when compared to the control, but at 72 h PI, the DI increased significantly (DI = 0.11 ± 0.03). Remarkably, dendritic beading in the somal compartment was observed at 24 h PI, indicative of dying back degeneration. When the injury level was increased (5% stretch, mild injury), microtubule fragmentation along the injured axons was observed, with a significant increase in DI at 24 h PI (DI = 0.17 ± 0.02) and 72 h PI (DI = 0.18 ± 0.01), relative to uninjured axons. The responses observed for both mild and very mild injuries are similar to those observed in the in vivo models of traumatic brain injury, suggesting that this model can be used to study neuronal trauma and will provide new insights into the cellular and molecular alterations characterizing the neuronal response to discrete axonal injury.
History
Journal
BiomicrofluidicsVolume
8Issue
4Article number
044110Publisher
AIP PublishingLocation
Melville, N.Y.Publisher DOI
ISSN
1932-1058Language
engPublication classification
C1.1 Refereed article in a scholarly journalCopyright notice
2014, AIP PublishingUsage metrics
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