Micro-Mechanics of Neuronal Compression II: The Cytoskeleton Strikes Back
Adam Fournier (Johns Hopkins), K.T. Ramesh (Johns Hopkins University)
Experimental Nanobiomechanics
Mon 9:00 - 10:30
Barus-Holley 163
Traumatic axon injury (TAI) can be characterized as focal or multi-focal damage to the white matter tracts of the central nervous system and has been linked to spinal cord injury (SCI) and traumatic brain injury (TBI). Degenerative responses to TAI have been linked to disruption of the cytoskeleton of neural axons. Our work utilizes a microfluidic compression platform, compatible with confocal microscopy, to successfully achieve visualization of the microtubule and neurofilament populations (cytoskeletal constituents of interest) prior to, during, and immediately after loading. We use primary hippocampal neurons from embryonic rats and fluorescently labeled cytoskeletal constituents through electroporation. Transmission electron microscopy (TEM) is conducted using the same experimental setup to gain insights regarding the dynamics of individual constituents of the cytoskeleton under focal loading. Control and focally compressed specimens are fixed and both cross sectional and axial slices are obtained immediately following loading. Comparisons between the volumes obtained through confocal imaging and the images from TEM help provide an understanding of the structural roles and responses of the cytoskeletal constituents under focal loading.