Longitudinal in vivo coherent anti-Stokes Raman scattering imaging of demyelination and remyelination in injured spinal cord.

In vivo imaging of white matter is important for the mechanistic understanding of demyelination and evaluation of remyelination therapies. Although white matter can be visualized by a strong coherent anti-Stokes Raman scattering (CARS) signal from axonal myelin, in vivo repetitive CARS imaging of the spinal cord remains a challenge due to complexities induced by the laminectomy surgery. We present a careful experimental design that enabled longitudinal CARS imaging of de- and remyelination at single axon level in live rats. In vivo CARS imaging of secretory phospholipase A(2) induced myelin vesiculation, macrophage uptake of myelin debris, and spontaneous remyelination by Schwann cells are sequentially monitored over a 3 week period. Longitudinal visualization of de- and remyelination at a single axon level provides a novel platform for rational design of therapies aimed at promoting myelin plasticity and repair.

[1]  D. Basso,et al.  A sensitive and reliable locomotor rating scale for open field testing in rats. , 1995, Journal of neurotrauma.

[2]  S. Whittemore,et al.  Functional and electrophysiological changes after graded traumatic spinal cord injury in adult rat , 2005, Experimental Neurology.

[3]  H. Bridge,et al.  High-resolution MRI: in vivo histology? , 2006, Philosophical Transactions of the Royal Society B: Biological Sciences.

[4]  Hans-Peter Hartung,et al.  Remyelinating strategies for the treatment of multiple sclerosis , 2002, Progress in Neurobiology.

[5]  T. B. Huff,et al.  In vivo coherent anti‐Stokes Raman scattering imaging of sciatic nerve tissue , 2007, Journal of microscopy.

[6]  B. Sabel,et al.  In vivo confocal neuroimaging (ICON) of CNS neurons , 1997, Nature Medicine.

[7]  Raymond P. Molloy,et al.  In vivo multiphoton microscopy of deep brain tissue. , 2004, Journal of neurophysiology.

[8]  A. Blight Delayed demyelination and macrophage invasion: a candidate for secondary cell damage in spinal cord injury. , 1985, Central nervous system trauma : journal of the American Paralysis Association.

[9]  Li Li,et al.  Quantitative coherent anti-Stokes Raman scattering imaging of lipid distribution in coexisting domains. , 2005, Biophysical journal.

[10]  Ji-Xin Cheng,et al.  Ex vivo and in vivo imaging of myelin fibers in mouse brain by coherent anti-Stokes Raman scattering microscopy. , 2008, Optics express.

[11]  M. Schwab Repairing the Injured Spinal Cord , 2002, Science.

[12]  G. Rougon,et al.  Quantitative analysis by in vivo imaging of the dynamics of vascular and axonal networks in injured mouse spinal cord , 2009, Proceedings of the National Academy of Sciences.

[13]  W. Klunk,et al.  Imaging of CNS myelin by positron-emission tomography. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[14]  R. Shi,et al.  Coherent anti‐stokes Raman scattering imaging of myelin degradation reveals a calcium‐dependent pathway in lyso‐PtdCho‐induced demyelination , 2007, Journal of neuroscience research.

[15]  Fred H. Gage,et al.  Therapeutic interventions after spinal cord injury , 2006, Nature Reviews Neuroscience.

[16]  H. Keirstead,et al.  Spinal cord injury is accompanied by chronic progressive demyelination , 2005, The Journal of comparative neurology.

[17]  Xiao-Ming Xu,et al.  Focal phospholipases A2 group III injections induce cervical white matter injury and functional deficits with delayed recovery concomitant with Schwann cell remyelination , 2007, Experimental Neurology.

[18]  M. Jacquin,et al.  Neuronal and Glial Apoptosis after Traumatic Spinal Cord Injury , 1997, The Journal of Neuroscience.

[19]  Jeff W Lichtman,et al.  In vivo imaging of axonal degeneration and regeneration in the injured spinal cord , 2005, Nature Medicine.

[20]  Riyi Shi,et al.  Glutamate Excitotoxicity Inflicts Paranodal Myelin Splitting and Retraction , 2009, PloS one.

[21]  Daniel Côté,et al.  Real-Time In Vivo Assessment of the Nerve Microenvironment with Coherent Anti–Stokes Raman Scattering Microscopy , 2009, Plastic and reconstructive surgery.