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Research Articles: Behavioral/Cognitive
Autophagy enhances memory erasure through synaptic destabilization
Mohammad Shehata1,2, Kareem Abdou1,2, Kiriko Choko1,2, Mina Matsuo3, Hirofumi Nishizono2,3 and Kaoru Inokuchi1,2
1Department of Biochemistry, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Toyama, 930-0194, Japan 2Japan Science and Technology Agency, CREST, University of Toyama, 930-0194, Japan 3Division of Animal Experimental Laboratory, Life Science Research Center, University of Toyama, Toyama, 930-0194, Japan
DOI: 10.1523/JNEUROSCI.3505-17.2018
Received: 10 December 2017
Revised: 14 February 2018
Accepted: 21 February 2018
Published: 19 March 2018
Author contributions: M.S. wrote the first draft of the paper; K.I. edited the paper. M.S., K.A., and K.I. designed research; M.S., K.A., and K.C. performed research; M.M., H.N., and K.I. contributed unpublished reagents/ analytic tools; M.S., K.A., K.C., and K.I. analyzed data; M.S. and K.I. wrote the paper.
Conflict of Interest: The authors declare no competing financial interests.
This work was supported by the Grant-in-Aid for Scientific Research on Innovative Areas “Memory dynamism” (JP25115002) from the Ministry of Education, Culture, Sports, Science, and Technology (MEXT), JSPS KAKENHI grant number JP23220009, the Core Research for Evolutional Science and Technology (CREST) program (JPMJCR13W1) of the Japan Science and Technology Agency (JST), the Mitsubishi Foundation, the Uehara Memorial Foundation, and the Takeda Science Foundation support to K.I.; and by a Grant-in-Aid for young scientists from JSPS KAKENHI grant number JP25830007 to M.S. The Otsuka Toshimi Scholarship Foundation supported K.A.
Corresponding authors: Kaoru Inokuchi, Ph.D., Professor, Department of Biochemistry, Faculty of Medicine, Graduate School of Medicine & Pharmaceutical Sciences, University of Toyama, 2630 Sugitani, Toyama 930-0194, Japan., E-mail: [email protected], Telephone: +81-76-434-7225; Fax: +81-76-434-5014, Mohammad Shehata, Ph.D., Visiting Associate, Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, 91125, USA. E-mail: [email protected]
Cite as: J. Neurosci ; 10.1523/JNEUROSCI.3505-17.2018
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Accepted manuscripts are peer-reviewed but have not been through the copyediting, formatting, or proofreading process.
Copyright © 2018 the authors
1 Title Page
2 Title: Autophagy enhances memory erasure through synaptic destabilization
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Autophagy contributes to fear memory destabilization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igure 2. Autophagy overcomes a reconsolidation-resistant boundary condition in an 860 AMPAR endocytosis-dependent manner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igure 3. Autophagy enhances fear memory destabilization in contextual fear conditioning 873 when targeted to amygdala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igure 4. 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