Autophagy-mediated stress response in motor neuron after transient ischemia in rabbits

Hironori Baba, Masahiro Sakurai, Koji Abe, Ryuji Tominaga

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Objective: Spinal cord injury is considered to be related to a vulnerability of spinal motor neurons to ischemia. However, the mechanisms underlying this vulnerability are not fully understood. We investigated the role of autophagy, which is an intracellular bulk degradation process, at motor neuron as a potential mechanism of neuronal death by immunohistochemical analysis for microtubule-associated protein light chain3 (LC3) and γ-aminobutyric-acid type-A-receptor-associated protein (GABARAP) which are considered as markers of autophagy. Methods: We used a rabbit spinal cord ischemia model with the use of a balloon catheter. The spinal cord was removed at 8 hours, 1, 2, or 7 days after 15 minutes of transient ischemia, and histologic changes were examined with hematoxylin-eosin staining. Western blot analysis for LC3 and GABARAP, temporal profiles of LC3 and GABARAP immunoreactivity, and double-label fluorescence immunocytochemical studies were performed. Results: In the ischemia group, about 85% of motor neurons were preserved until 2 days after reperfusion, but were selectively lost at 7 days (P <.001 compared with sham group). Western blot analysis demonstrated slight immunoreactivity for LC3 and GABARAP in the sham-operated spinal cords. In contrast, the ischemia group LC3 and GABARAP immunoreactivity became apparent at 8 hours after reperfusion. With quantitative analysis we found that ischemia affected expression profiles of LC3-II and GABARAP. At 8 hours after reperfusion, co-labeling of LC3 and GABARAP were observed in the same motor neurons that eventually died. Conclusion: These data suggest that autophagy was induced in motor neurons by transient spinal cord ischemia in rabbits.

Original languageEnglish
Pages (from-to)381-387
Number of pages7
JournalJournal of Vascular Surgery
Volume50
Issue number2
DOIs
Publication statusPublished - Aug 2009

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Autophagy
Motor Neurons
Ischemia
Rabbits
Light
Spinal Cord Ischemia
Reperfusion
Spinal Cord
Western Blotting
Aminobutyrates
Microtubule-Associated Proteins
Hematoxylin
Eosine Yellowish-(YS)
Spinal Cord Injuries
Catheters
Fluorescence
Staining and Labeling
Proteins

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine
  • Surgery

Cite this

Autophagy-mediated stress response in motor neuron after transient ischemia in rabbits. / Baba, Hironori; Sakurai, Masahiro; Abe, Koji; Tominaga, Ryuji.

In: Journal of Vascular Surgery, Vol. 50, No. 2, 08.2009, p. 381-387.

Research output: Contribution to journalArticle

Baba, Hironori ; Sakurai, Masahiro ; Abe, Koji ; Tominaga, Ryuji. / Autophagy-mediated stress response in motor neuron after transient ischemia in rabbits. In: Journal of Vascular Surgery. 2009 ; Vol. 50, No. 2. pp. 381-387.
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abstract = "Objective: Spinal cord injury is considered to be related to a vulnerability of spinal motor neurons to ischemia. However, the mechanisms underlying this vulnerability are not fully understood. We investigated the role of autophagy, which is an intracellular bulk degradation process, at motor neuron as a potential mechanism of neuronal death by immunohistochemical analysis for microtubule-associated protein light chain3 (LC3) and γ-aminobutyric-acid type-A-receptor-associated protein (GABARAP) which are considered as markers of autophagy. Methods: We used a rabbit spinal cord ischemia model with the use of a balloon catheter. The spinal cord was removed at 8 hours, 1, 2, or 7 days after 15 minutes of transient ischemia, and histologic changes were examined with hematoxylin-eosin staining. Western blot analysis for LC3 and GABARAP, temporal profiles of LC3 and GABARAP immunoreactivity, and double-label fluorescence immunocytochemical studies were performed. Results: In the ischemia group, about 85{\%} of motor neurons were preserved until 2 days after reperfusion, but were selectively lost at 7 days (P <.001 compared with sham group). Western blot analysis demonstrated slight immunoreactivity for LC3 and GABARAP in the sham-operated spinal cords. In contrast, the ischemia group LC3 and GABARAP immunoreactivity became apparent at 8 hours after reperfusion. With quantitative analysis we found that ischemia affected expression profiles of LC3-II and GABARAP. At 8 hours after reperfusion, co-labeling of LC3 and GABARAP were observed in the same motor neurons that eventually died. Conclusion: These data suggest that autophagy was induced in motor neurons by transient spinal cord ischemia in rabbits.",
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