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

International Journal of Molecular Sciences

Hideki Horiuchi³  Yusuke Oshima²  Tadanori Ogata³  Tadao Morino³  Seiji Matsuda¹  Hiromasa Miura³  Takeshi Imamura² 

[1] Department of Anatomy and Embryology, Ehime University Graduate School of Medicine, Ehime 791-0295, Japan; E-Mail:;Department of Molecular Medicine for Pathogenesis, Ehime University Graduate School of Medicine, Ehime 791-0295, Japan; E-Mail:;Department of Orthopaedic Surgery, Ehime University Graduate School of Medicine, Ehime 791-0295, Japan; E-Mails:

关键词: spinal cord injury;    axon degeneration;    multiphoton excitation microscope;    transgenic mouse;   

DOI  :  10.3390/ijms160715785

来源: mdpi

【 摘 要 】

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.

【 授权许可】

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