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Int. J. Mol. Sci. 2015, 16(7), 15785-15799; doi:10.3390/ijms160715785

Evaluation of Injured Axons Using Two-Photon Excited Fluorescence Microscopy after Spinal Cord Contusion Injury in YFP-H Line Mice

1
Department of Orthopaedic Surgery, Ehime University Graduate School of Medicine, Ehime 791-0295, Japan
2
Department of Molecular Medicine for Pathogenesis, Ehime University Graduate School of Medicine, Ehime 791-0295, Japan
3
Translational Research Center, Ehime University Hospital, Ehime 791-0295, Japan
4
Division of Bio-imaging, Proteo-Science Center, Ehime University, Ehime 791-0295, Japan
5
Department of Anatomy and Embryology, Ehime University Graduate School of Medicine, Ehime 791-0295, Japan
These authors contributed equally to this work.
*
Authors to whom correspondence should be addressed.
Academic Editor: Xiaofeng Jia
Received: 24 March 2015 / Revised: 2 July 2015 / Accepted: 8 July 2015 / Published: 13 July 2015
(This article belongs to the Special Issue Neurological Injuries’ Monitoring, Tracking and Treatment)
View Full-Text   |   Download PDF [5464 KB, uploaded 13 July 2015]   |  

Abstract

Elucidation of the process of degeneration of injured axons is important for the development of therapeutic modules for the treatment of spinal cord injuries. The aim of this study was to establish a method for time-lapse observation of injured axons in living animals after spinal cord contusion injury. YFP (yellow fluorescent protein)-H transgenic mice, which we used in this study, express fluorescence in their nerve fibers. Contusion damage to the spinal cord at the 11th vertebra was performed by IH (Infinite Horizon) impactor, which applied a pressure of 50 kdyn. The damaged spinal cords were re-exposed during the observation period under anesthesia, and then observed by two-photon excited fluorescence microscopy, which can observe deep regions of tissues including spinal cord axons. No significant morphological change of injured axons was observed immediately after injury. Three days after injury, the number of axons decreased, and residual axons were fragmented. Seven days after injury, only fragments were present in the damaged tissue. No hind-limb movement was observed during the observation period after injury. Despite the immediate paresis of hind-limbs following the contusion injury, the morphological degeneration of injured axons was delayed. This method may help clarification of pathophysiology of axon degeneration and development of therapeutic modules for the treatment of spinal cord injury. View Full-Text
Keywords: spinal cord injury; axon degeneration; multiphoton excitation microscope; transgenic mouse spinal cord injury; axon degeneration; multiphoton excitation microscope; transgenic mouse
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MDPI and ACS Style

Horiuchi, H.; Oshima, Y.; Ogata, T.; Morino, T.; Matsuda, S.; Miura, H.; Imamura, T. Evaluation of Injured Axons Using Two-Photon Excited Fluorescence Microscopy after Spinal Cord Contusion Injury in YFP-H Line Mice. Int. J. Mol. Sci. 2015, 16, 15785-15799.

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