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Hypoxia-Induced Neuroinflammation and Learning–Memory Impairments in Adult Zebrafish Are Suppressed by Glucosamine

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Abstract

This study investigated changes in neuroinflammation and cognitive function in adult zebrafish exposed to acute hypoxia and protective effects of glucosamine (GlcN) against hypoxia-induced brain damage. The survival rate of zebrafish following exposure to hypoxia was improved by GlcN pretreatment. Moreover, hypoxia-induced upregulation of neuroglobin, NOS2α, glial fibrillary acidic protein, and S100β in zebrafish was suppressed by GlcN. Hypoxia stimulated cell proliferation in the telencephalic ventral domain and in cerebellum subregions. GlcN decreased the number of bromodeoxyuridine (BrdU)-positive cells in the telencephalon region, but not in cerebellum regions. Transient motor neuron defects, assessed by measuring the locomotor and exploratory activity of zebrafish exposed to hypoxia recovered quickly. GlcN did not affect hypoxia-induced motor activity changes. In passive avoidance tests, hypoxia impaired learning and memory ability, deficits that were rescued by GlcN. A learning stimulus increased the nuclear translocation of phosphorylated cAMP response element binding protein (p-CREB), an effect that was greatly inhibited by hypoxia. GlcN restored nuclear p-CREB after a learning trial in hypoxia-exposed zebrafish. The neurotransmitters, γ-aminobutyric acid and glutamate, were increased after hypoxia in the zebrafish brain, and GlcN further increased their levels. In contrast, acetylcholine levels were reduced by hypoxia and restored by GlcN. Acetylcholinesterase inhibitor physostigmine partially reversed the impaired learning and memory of hypoxic zebrafish. This study represents the first examination of the molecular mechanisms underlying hypoxia-induced memory and learning defects in a zebrafish model. Our results further suggest that GlcN-associated hexosamine metabolic pathway could be an important therapeutic target for hypoxic brain damage.

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Funding

This work was supported by the National Research Foundation (NRF) of Korea grants, NRF-2017R1A2B2007199, NRF-2017R1D1A1B03031431, and the Brain Research Program 2015M3C7A1028373 and WCSL (World Class Smart Lab) research grant of Inha University.

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  1. Y Lee and S Lee equally contributed to this work. IO Han and CJ Lee equally contributed to this work

Authors and Affiliations

  1. Department of Physiology and Biophysics, College of Medicine, Inha University, 100 Inha Ro, Nam-gu, Incheon, 22212, South Korea

    Yunkyoung Lee, Ji-Won Park, Ji-Sun Hwang, Sang-Min Kim & Inn-Oc Han

  2. Department of Biological Sciences, Inha University, Incheon, South Korea

    Sujeong Lee & Chang-Joong Lee

  3. Department of Brain and Cognitive Sciences, Ewha Brain Institute, College of Pharmacy, Ewha Womans University, Seoul, South Korea

    In Kyoon Lyoo

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  1. Yunkyoung Lee
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Contributions

IO Han designed the study and supervised the project; Y Lee and S Lee wrote the manuscript and analyzed the data; JW Park, SM Kim and JS Hwang performed experiments and the imaging studies; CJ Lee and IK Lyoo contributed to designing experimental procedures. All authors reviewed the manuscript.

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Correspondence to Chang-Joong Lee or Inn-Oc Han.

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Supplemental Figure 1

Change of O-GlcNAc level in the brain of adult zebrafish in response to hypoxia with or without GlcN pretreatment. Adult zebrafish were pretreated with 1 g/L GlcN for 12 h and then exposed to hypoxic conditions for 8 min. After a 6- or 24-h recovery period, total protein extraction from brain were prepared and O-GlcNAc protein level was determined by Western blotting along with ⍺-Tubulin as a loading control. Shown blots are representatives of three independent experiments. All values are means ± SEM. *P < 0.05 versus C; #P < 0.05 versus H. A.U., arbitrary units; C, control (normoxia); H, hypoxia; G, GlcN; HG, hypoxia + GlcN. (GIF 45 kb)

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Lee, Y., Lee, S., Park, JW. et al. Hypoxia-Induced Neuroinflammation and Learning–Memory Impairments in Adult Zebrafish Are Suppressed by Glucosamine. Mol Neurobiol 55, 8738–8753 (2018). https://doi.org/10.1007/s12035-018-1017-9

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