Effects of N-acetylcysteine amide on anxiety and stress behavior in zebrafish
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Anxiety disorders are highly prevalent and a leading cause of disability worldwide. Their etiology is related to stress, an adaptive response of the organism to restore homeostasis, in which oxidative stress and glutamatergic hyperactivity are involved. N-Acetylcysteine (NAC) is a multitarget approved drug proved to be beneficial in the treatment of various mental disorders. Nevertheless, NAC has low membrane permeability and poor bioavailability and its limited delivery to the brain may explain inconsistencies in the literature. N-Acetylcysteine amide (AD4) is a synthetic derivative of NAC in which the carboxyl group was modified to an amide. The amidation of AD4 improved lipophilicity and blood-brain barrier permeability and enhanced its antioxidant properties. The purpose of this study was to investigate the effects of AD4 on behavioral and biochemical parameters in zebrafish anxiety models. Neither AD4 nor NAC induced effects on locomotion and anxiety-related parameters in the novel tank test. However, in the light/dark test, AD4 (0.001 mg/L) increased the time spent in the lit side in a concentration 100 times lower than NAC (0.1 mg/L). In the acute restraint stress protocol, NAC and AD4 (0.001 mg/L) showed anxiolytic properties without meaningful effects on oxidative status. The study suggests that AD4 has anxiolytic effects in zebrafish with higher potency than the parent compound. Additional studies are warranted to characterize the anxiolytic profile of AD4 and its potential in the management of anxiety disorders.
KeywordsN-Acetylcysteine amide Anxiety Acute restraint stress AD4 NACA
We thank Prof. Daphne Atlas from Hebrew University of Jerusalem for providing AD4 and encouraging this study and Brandon Guinalli for helping with the figure designs.
CR designed and performed the experiments, analyzed the data, prepared figures and tables, and authored and reviewed drafts of the paper. RM, RB, MM, and AS performed the experiments and reviewed drafts of the paper. AH, EE, and AP designed the experiments, analyzed the data, contributed reagents/materials/analysis tools, prepared figures and tables, and authored and reviewed drafts of the paper. All authors read and approved the manuscript.
This work was supported by CNPq (302800/2017-4) and FAPERGS (#17/2551-0000974-6); CGR is recipient of a fellowship from CAPES and AP from CNPq.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflicts of interest.
All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.
- Bahat-Stroomza M, Gilgun-Sherki Y, Offen D, Panet H, Saada A, Krool-Galron N, Barzilai A, Atlas D, Melamed E (2005) A novel thiol antioxidant that crosses the blood brain barrier protects dopaminergic neurons in experimental models of Parkinson’s disease. Eur J Neurosci 21:637–646. https://doi.org/10.1111/j.1460-9568.2005.03889.x CrossRefPubMedGoogle Scholar
- Bystritsky A, Khalsa SS, Cameron ME, Schiffman J (2013) Current diagnosis and treatment of anxiety disorders. Pharm Ther 38:30–57Google Scholar
- Egan RJ, Bergner CL, Hart PC, Cachat JM, Canavello PR, Elegante MF, Elkhayat SI, Bartels BK, Tien AK, Tien DH, Mohnot S, Beeson E, Glasgow E, Amri H, Zukowska Z, Kalueff AV (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
- Grinberg L, Fibach E, Amer J, Atlas D (2005) N-acetylcysteine amide, a novel cell-permeating thiol, restores cellular glutathione and protects human red blood cells from oxidative stress. Free Radic Biol Med 38:136–145. https://doi.org/10.1016/j.freeradbiomed.2004.09.025 CrossRefPubMedGoogle Scholar
- Koltunowska D, Gibula-Bruzda E, Kotlinska JH (2013) The influence of ionotropic and metabotropic glutamate receptor ligands on anxiety-like effect of amphetamine withdrawal in rats. Prog Neuropsychopharmacol Biol Psychiatry 45:242–249. https://doi.org/10.1016/j.pnpbp.2013.04.013 CrossRefPubMedGoogle Scholar
- Kowalczyk-Pachel D, Iciek M, Wydra K, Nowak E, Górny M, Filip M, Włodek L, Lorenc-Koci E (2016) Cysteine metabolism and oxidative processes in the rat liver and kidney after acute and repeated cocaine treatment. PLOS ONE 11:e0147238. https://doi.org/10.1371/journal.pone.0147238 CrossRefPubMedPubMedCentralGoogle Scholar
- Kysil EV, Meshalkina DA, Frick EE, Echevarria DJ, Rosemberg DB, Maximino C, Lima MG, Abreu MS, Giacomini AC, Barcellos LJG, Song C, Kalueff AV (2017) Comparative analyses of zebrafish anxiety-like behavior using conflict-based novelty tests. Zebrafish 14:197–208. https://doi.org/10.1089/zeb.2016.1415 CrossRefPubMedGoogle Scholar
- Marcon M, Herrmann AP, Mocelin R, Rambo CL, Koakoski G, Abreu MS, Conterato GM, Kist LW, Bogo MR, Zanatta L, Barcellos LJ, Piato AL (2016) Prevention of unpredictable chronic stress-related phenomena in zebrafish exposed to bromazepam, fluoxetine and nortriptyline. Psychopharmacology (Berl) 233:3815–3824. https://doi.org/10.1007/s00213-016-4408-5 CrossRefGoogle Scholar
- Minarini A, Ferrari S, Galletti M, Giambalvo N, Perrone D, Rioli G, Galeazzi GM (2017) N-acetylcysteine in the treatment of psychiatric disorders: current status and future prospects. Expert Opin Drug Metab Toxicol 13:279–292. https://doi.org/10.1080/17425255.2017.1251580 CrossRefPubMedGoogle Scholar
- Mocelin R, Marcon M, D’ambros S, Mattos J, Sachett A, Siebel AM, Herrmann AP, Piato A (2018b) N-Acetylcysteine reverses anxiety and oxidative damage induced by unpredictable chronic stress in zebrafish. Mol Neurobiol. 56:1188–1195. https://doi.org/10.1007/s12035-018-1165-y CrossRefPubMedGoogle Scholar
- Offen D, Gilgun-Sherki Y, Barhum Y, Benhar M, Grinberg L, Reich R, Melamed E, Atlas D (2004) A low molecular weight copper chelator crosses the blood-brain barrier and attenuates experimental autoimmune encephalomyelitis. J Neurochem 89:1241–1251. https://doi.org/10.1111/j.1471-4159.2004.02428.x CrossRefPubMedGoogle Scholar
- Pandya JD, Readnower RD, Patel SP, Yonutas HM, Pauly JR, Goldstein GA, Rabchevsky AG, Sullivan PG (2014) N-acetylcysteine amide confers neuroprotection, improves bioenergetics and behavioral outcome following TBI. Exp Neurol 257:106–113. https://doi.org/10.1016/j.expneurol.2014.04.020 CrossRefPubMedPubMedCentralGoogle Scholar
- Piato AL, Rosemberg DB, Capiotti KM, Siebel AM, Herrmann AP, Ghisleni G, Vianna MR, Bogo MR, Lara DR, Bonan CD (2011) Acute restraint stress in zebrafish: behavioral parameters and purinergic signaling. Neurochem Res 36:1876–1886. https://doi.org/10.1007/s11064-011-0509-z CrossRefPubMedGoogle Scholar
- Quadros VA, Silveira A, Giuliani GS, Didonet F, Silveira AS, Nunes ME, Silva TO, Loro VL, Rosemberg DB (2016) Strain- and context-dependent behavioural responses of acute alarm substance exposure in zebrafish. Behav Processes 122:1–11. https://doi.org/10.1016/j.beproc.2015.10.014 CrossRefPubMedGoogle Scholar
- Stewart A, Wu N, Cachat J, Hart P, Gaikwad S, Wong K, Utterback E, Gilder T, Kyzar E, Newman A, Carlos D, Chang K, Hook M, Rhymes C, Caffery M, Greenberg M, Zadina J, Kalueff AV (2011) Pharmacological modulation of anxiety-like phenotypes in adult zebrafish behavioral models. Prog Neuropsychopharmacol Biol Psychiatry 35:1421–1431. https://doi.org/10.1016/j.pnpbp.2010.11.035 CrossRefPubMedGoogle Scholar
- Zhang X, Banerjee A, Banks WA, Ercal N (2009) N-Acetylcysteine amide protects against methamphetamine-induced oxidative stress and neurotoxicity in immortalized human brain endothelial cells. Brain Res 1275:87–95. https://doi.org/10.1016/j.brainres.2009.04.008 CrossRefPubMedPubMedCentralGoogle Scholar
- Zhou Y, Wang H, Zhou X et al (2018) N-acetylcysteine amide provides neuroprotection via Nrf2-ARE pathway in a mouse model of traumatic brain injury. Drug Des. Devel. Ther, In https://www.dovepress.com/article_metric.php?article_id=42804.CrossRefGoogle Scholar