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Inactivation of NSF ATPase Leads to Cathepsin B Release After Transient Cerebral Ischemia

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A Correction to this article was published on 23 May 2018

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Abstract

Neurons have extraordinary large cell membrane surface area, thus requiring extremely high levels of intracellular membrane-trafficking activities. Consequently, defects in the membrane-trafficking activities preferentially affect neurons. A critical molecule for controlling the membrane-trafficking activities is the N-ethylmaleimide-sensitive factor (NSF) ATPase. This study is to investigate the cascade of events of NSF ATPase inactivation, resulting in a massive buildup of late endosomes (LEs) and fatal release of cathepsin B (CTSB) after transient cerebral ischemia using the 2-vessel occlusion with hypotension (2VO+Hypotension) global brain ischemia model. Rats were subjected to 20 min of transient cerebral ischemia followed by 0.5, 4, 24, and 72 h of reperfusion. Neuronal histopathology and ultrastructure were examined by the light and electron microscopy, respectively. Western blotting and confocal microscopy were utilized for analyzing the levels, redistribution, and co-localization of Golgi apparatus and endosome or lysosome markers. Transient cerebral ischemia leads to delayed neuronal death that occurs at 48–72 h of reperfusion mainly in hippocampal CA1 and neocortical (Cx) layers 3 and 5 pyramidal neurons. During the delayed period, NSF ATPase is irreversibly trapped into inactive protein aggregates selectively in post-ischemic neurons destined to die. NSF inactivation leads to a massive buildup of Golgi fragments, transport vesicles (TVs) and late endosomes (LEs), and release of the 33 kDa LE type of CTSB, which is followed by delayed neuronal death after transient cerebral ischemia. The results support a novel hypothesis that transient cerebral ischemia leads to NSF inactivation, resulting in a cascade of events of fatal release of CTSB and delayed neuronal death after transient cerebral ischemia.

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Change history

  • 23 May 2018

    The author name “Dr. Jiang Wu” needed to be added as the 3rd author. The author institutional affiliations are correspondingly adjusted. The authors regret these errors.

    The original article has been corrected.

Abbreviations

NSF:

N-ethylmaleimide sensitive factor ATPase

SNAREs:

Soluble NSF attachment protein receptors

SNAP:

Soluble NSF attachment protein

CTSB:

Cathepsin B

TVs:

Transport vesicles

LE:

Late endosome

EL:

Endolysosome

L:

Lysosome

MOMP:

Mitochondrial outer membrane permeabilization

IRI:

Ischemia-reperfusion injury

DG:

Dentate gyrus

EM:

Electron microscopy

Vti1b:

Vesicle transport through interaction with t-SNAREs homolog 1B

TGN38:

Trans-Golgi network membrane protein 38 kDa

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Funding

This work was supported by National Institutes of Health (NIH) grants: NS36810, NS40407, and NS097875; by Veteran Affair Merit grant: I01BX001696; and by the American Heart Association 0940042N-5 to B.R.H.

Author information

Authors and Affiliations

  1. Department of Neurology, The First Teaching Hospital, Jilin University, Changchun, China

    Dong Yuan & Jiang Wu

  2. Department of Anesthesiology and Neurology, Shock Trauma and Anesthesiology Research Center, University of Maryland School of Medicine, Baltimore, MD, 21201, USA

    Dong Yuan, Chunli Liu & Bingren Hu

  3. Veterans Affairs Maryland Health Center System, 10 North Greene Street, Baltimore, MD, 21201, USA

    Bingren Hu

Authors
  1. Dong Yuan
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  2. Chunli Liu
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  3. Jiang Wu
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Correspondence to Bingren Hu.

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Conflict of Interest

Dong Yuan, Chunli Liu, and Bingren Hu declare no conflict of interest.

Ethical Approval

This article does not contain any studies with human subjects. All the experimental procedures involving using animals were approved by the Animal Use and Care Committee in the University of Maryland School of Medicine.

Additional information

The original version of this article was revised: The author name “Dr. Jiang Wu” needed to be added as the 3rd author.

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Yuan, D., Liu, C., Wu, J. et al. Inactivation of NSF ATPase Leads to Cathepsin B Release After Transient Cerebral Ischemia. Transl. Stroke Res. 9, 201–213 (2018). https://doi.org/10.1007/s12975-017-0571-1

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  • DOI: https://doi.org/10.1007/s12975-017-0571-1

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