Fig. 1

Using microfluidic chamber devices to model CNS axon regeneration and degeneration after injury and ischemia. A Schematic of a microfluidic chamber device that physically and fluidically separates axons from soma-dendritic area of neurons. Cortical neurons were plated in the soma-dendritic chamber (yellow) at DIV0, and only axons grow into the axon terminal chamber (purple) through the 450-μm microgrooves (red box). B Co-immunostaining validates axon compartments (labeled by βIII-tubulin) and soma-dendritic compartments (labeled by MAP2). C, D Schematic showing axotomy via vacuum aspiration within axon chambers (C) and representative images of axon chambers showing relative axonal regrowth capacity 6 days post injury (D). WT or snph KO cortical neurons were axotomized at DIV10 and labeled with βIII-tubulin for imaging 6 days post axotomy. E, F Schematic showing OGD-R treatment within axonal chamber (E) and representative images of axon chambers showing axonal integrity before (left) or after (right) OGD-R treatment. Cortical neurons at DIV14 were treated with OGD for 30 minutes and reperfusion for 8 hours, followed by immunostaining with βIII-tubulin. Note that the ischemic stress induces bead-like or fragmented axons. Scale bars, 50 μm (B), 100 μm (D), 20 μm (F). Imaging (B) is adapted from Farfel-Becker et al. 2019 with permission