Hypoxia-Induced Neuroinflammation and Learning–Memory Impairments in Adult Zebrafish Are Suppressed by Glucosamine
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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.
KeywordsHypoxia Glucosamine Neuroinflammation CREB Zebrafish
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.
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.
Compliance with Ethical Standards
Conflict of Interest
The authors declare that they have no conflicts of interest.
- 3.Braga MM, Rico EP, Cordova SD, Pinto CB, Blaser RE, Dias RD, Rosemberg DB, Oliveira DL et al (2013) Evaluation of spontaneous recovery of behavioral and brain injury profiles in zebrafish after hypoxia. Behav Brain Res 253:145–151. https://doi.org/10.1016/j.bbr.2013.07.019 CrossRefPubMedGoogle Scholar
- 4.Braga MM, Silva ES, Moraes TB, Schirmbeck GH, Rico EP, Pinto CB, Rosemberg DB, Dutra-Filho CS et al (2016) Brain zinc chelation by diethyldithiocarbamate increased the behavioral and mitochondrial damages in zebrafish subjected to hypoxia. Sci Rep 6:20279. https://doi.org/10.1038/srep20279 CrossRefPubMedPubMedCentralGoogle Scholar
- 8.Altman J (1969) Autoradiographic and histological studies of postnatal neurogenesis. IV. Cell proliferation and migration in the anterior forebrain, with special reference to persisting neurogenesis in the olfactory bulb. J Comp Neurol 137:433–457. https://doi.org/10.1002/cne.901370404 CrossRefPubMedGoogle Scholar
- 14.Liu J, Pang Y, Chang T, Bounelis P, Chatham JC, Marchase RB (2006) Increased hexosamine biosynthesis and protein O-GlcNAc levels associated with myocardial protection against calcium paradox and ischemia. J Mol Cell Cardiol 40:303–312. https://doi.org/10.1016/j.yjmcc.2005.11.003 CrossRefPubMedGoogle Scholar
- 17.Algra SO, Groeneveld KM, Schadenberg AW, Haas F, Evens FC, Meerding J, Koenderman L, Jansen NJ et al (2013) Cerebral ischemia initiates an immediate innate immune response in neonates during cardiac surgery. J Neuroinflammation 10:24. https://doi.org/10.1186/1742-2094-10-24 CrossRefPubMedPubMedCentralGoogle Scholar
- 21.Kida S, Serita T (2014) Functional roles of CREB as a positive regulator in the formation and enhancement of memory. Brain Res Bull 105:17–24. https://doi.org/10.1016/j.brainresbull.2014.04.011 CrossRefPubMedGoogle Scholar
- 25.Trifilieff P, Herry C, Vanhoutte P, Caboche J, Desmedt A, Riedel G, Mons N, Micheau J (2006) Foreground contextual fear memory consolidation requires two independent phases of hippocampal ERK/CREB activation. Learn Mem 13:349–358. https://doi.org/10.1101/lm.80206 CrossRefPubMedPubMedCentralGoogle Scholar
- 27.Cammarota M, Bevilaqua LR, Ardenghi P, Paratcha G, Levi de Stein M, Izquierdo I, Medina JH (2000) Learning-associated activation of nuclear MAPK, CREB and Elk-1, along with Fos production, in the rat hippocampus after a one-trial avoidance learning: abolition by NMDA receptor blockade. Brain Res Mol Brain Res 76:36–46CrossRefGoogle Scholar
- 28.Tropea TF, Kosofsky BE, Rajadhyaksha AM (2008) Enhanced CREB and DARPP-32 phosphorylation in the nucleus accumbens and CREB, ERK, and GluR1 phosphorylation in the dorsal hippocampus is associated with cocaine-conditioned place preference behavior. J Neurochem 106:1780–1790. https://doi.org/10.1111/j.1471-4159.2008.05518.x CrossRefPubMedGoogle Scholar
- 29.Barros TP, Alderton WK, Reynolds HM, Roach AG, Berghmans S (2008) Zebrafish: an emerging technology for in vivo pharmacological assessment to identify potential safety liabilities in early drug discovery. Br J Pharmacol 154:1400–1413. https://doi.org/10.1038/bjp.2008.249 CrossRefPubMedPubMedCentralGoogle Scholar
- 30.Senger MR, Rosemberg DB, Rico EP, de Bem Arizi M, Dias RD, Bogo MR, Bonan CD (2006) In vitro effect of zinc and cadmium on acetylcholinesterase and ectonucleotidase activities in zebrafish (Danio rerio) brain. Toxicol in Vitro 20:954–958. https://doi.org/10.1016/j.tiv.2005.12.002 CrossRefPubMedGoogle Scholar
- 31.Egan RJ, Bergner CL, Hart PC, Cachat JM, Canavello PR, Elegante MF, Elkhayat SI, Bartels BK et al (2009) Understanding behavioral and physiological phenotypes of stress and anxiety in zebrafish. Behav Brain Res 205:38–44. https://doi.org/10.1016/j.bbr.2009.06.022 CrossRefPubMedPubMedCentralGoogle Scholar
- 45.Champattanachai V, Marchase RB, Chatham JC (2008) Glucosamine protects neonatal cardiomyocytes from ischemia-reperfusion injury via increased protein O-GlcNAc and increased mitochondrial Bcl-2. Am J Physiol Cell Physiol 294:C1509–C1520. https://doi.org/10.1152/ajpcell.00456.2007 CrossRefPubMedPubMedCentralGoogle Scholar
- 52.Aviles-Reyes RX, Angelo MF, Villarreal A, Rios H, Lazarowski A, Ramos AJ (2010) Intermittent hypoxia during sleep induces reactive gliosis and limited neuronal death in rats: implications for sleep apnea. J Neurochem 112:854–869. https://doi.org/10.1111/j.1471-4159.2009.06535.x CrossRefPubMedGoogle Scholar