A novel in vitro injury model based on microcontact printing demonstrates negative effects of hydrogen peroxide on axonal regeneration both in absence and presence of glia.
2013 (English)In: Journal of Neurotrauma, ISSN 0897-7151, E-ISSN 1557-9042, Vol. 30, no 5, 392-402 p.Article in journal (Refereed) Published
The molecular processes involved in axonal regeneration following traumatic brain injury (TBI) are still not fully understood. In this study we have established a novel in vitro injury model of TBI based on microcontact printing (µCP) that enables close up investigations of injured neurons. The model is also suitable for quantitative measurements of axonal outgrowth, making it a useful tool in the studies of basic mechanisms behind axonal regeneration. Cortical neurons from mouse embryos are cultured on µCP cover slips for 8 days and the neurons are then injured in a precise manner using a thin plastic tip that do not affect the µCP pattern of extracellular matrix proteins. By close-up time-lapse experiments and immunostainings we show that the neurons have a tremendous capacity to regenerate their neurites after injury. The cut induces growth cone formation and the regenerating axons strictly follow the µCP pattern. Moreover, by using the injury model we demonstrate that hydrogen peroxide (H2O2) decreases axonal regeneration after injury without affecting the neurons ability to form growth cones. Co-culture with glial cells does not rescue the axonal regeneration, indicating that the mechanism by which H2O2 affects axonal regeneration differ from its cytotoxic effect.
Place, publisher, year, edition, pages
2013. Vol. 30, no 5, 392-402 p.
Medical and Health Sciences
IdentifiersURN: urn:nbn:se:uu:diva-188234DOI: 10.1089/neu.2012.2562ISI: 000315891600009PubMedID: 23057993OAI: oai:DiVA.org:uu-188234DiVA: diva2:576915