Evaluation of Injured Axons Using Two-Photon Excited Fluorescence Microscopy after Spinal Cord Contusion Injury in YFP-H Line Mice
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 in...
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doaj-c39a22966bb24e888367efcaf773bae42020-11-24T22:16:08ZengMDPI AGInternational Journal of Molecular Sciences1422-00672015-07-01167157851579910.3390/ijms160715785ijms160715785Evaluation of Injured Axons Using Two-Photon Excited Fluorescence Microscopy after Spinal Cord Contusion Injury in YFP-H Line MiceHideki Horiuchi0Yusuke Oshima1Tadanori Ogata2Tadao Morino3Seiji Matsuda4Hiromasa Miura5Takeshi Imamura6Department of Orthopaedic Surgery, Ehime University Graduate School of Medicine, Ehime 791-0295, JapanDepartment of Molecular Medicine for Pathogenesis, Ehime University Graduate School of Medicine, Ehime 791-0295, JapanDepartment of Orthopaedic Surgery, Ehime University Graduate School of Medicine, Ehime 791-0295, JapanDepartment of Orthopaedic Surgery, Ehime University Graduate School of Medicine, Ehime 791-0295, JapanDepartment of Anatomy and Embryology, Ehime University Graduate School of Medicine, Ehime 791-0295, JapanDepartment of Orthopaedic Surgery, Ehime University Graduate School of Medicine, Ehime 791-0295, JapanDepartment of Molecular Medicine for Pathogenesis, Ehime University Graduate School of Medicine, Ehime 791-0295, JapanElucidation 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.http://www.mdpi.com/1422-0067/16/7/15785spinal cord injuryaxon degenerationmultiphoton excitation microscopetransgenic mouse |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Hideki Horiuchi Yusuke Oshima Tadanori Ogata Tadao Morino Seiji Matsuda Hiromasa Miura Takeshi Imamura |
spellingShingle |
Hideki Horiuchi Yusuke Oshima Tadanori Ogata Tadao Morino Seiji Matsuda Hiromasa Miura Takeshi Imamura 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 spinal cord injury axon degeneration multiphoton excitation microscope transgenic mouse |
author_facet |
Hideki Horiuchi Yusuke Oshima Tadanori Ogata Tadao Morino Seiji Matsuda Hiromasa Miura Takeshi Imamura |
author_sort |
Hideki Horiuchi |
title |
Evaluation of Injured Axons Using Two-Photon Excited Fluorescence Microscopy after Spinal Cord Contusion Injury in YFP-H Line Mice |
title_short |
Evaluation of Injured Axons Using Two-Photon Excited Fluorescence Microscopy after Spinal Cord Contusion Injury in YFP-H Line Mice |
title_full |
Evaluation of Injured Axons Using Two-Photon Excited Fluorescence Microscopy after Spinal Cord Contusion Injury in YFP-H Line Mice |
title_fullStr |
Evaluation of Injured Axons Using Two-Photon Excited Fluorescence Microscopy after Spinal Cord Contusion Injury in YFP-H Line Mice |
title_full_unstemmed |
Evaluation of Injured Axons Using Two-Photon Excited Fluorescence Microscopy after Spinal Cord Contusion Injury in YFP-H Line Mice |
title_sort |
evaluation of injured axons using two-photon excited fluorescence microscopy after spinal cord contusion injury in yfp-h line mice |
publisher |
MDPI AG |
series |
International Journal of Molecular Sciences |
issn |
1422-0067 |
publishDate |
2015-07-01 |
description |
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. |
topic |
spinal cord injury axon degeneration multiphoton excitation microscope transgenic mouse |
url |
http://www.mdpi.com/1422-0067/16/7/15785 |
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