It is known that continuous abuse of amphetamine (AMPH) results in alterations in neuronal structure and cognitive behaviors related to the reward system. However, the impact of AMPH abuse on the hippocampus remains unknown. The aim of this study was to determine the damage caused by AMPH in the hippocampus in an addiction model. We reproduced the AMPH sensitization model proposed by Robinson et al. in 1997 and performed the novel object recognition test (NORt) to evaluate learning and memory behaviors. After the NORt, we performed Golgi–Cox staining, a stereological cell count, immunohistochemistry to determine the presence of GFAP, CASP3, and MT-III, and evaluated oxidative stress in the hippocampus. We found that AMPH treatment generates impairment in short- and long-term memories and a decrease in neuronal density in the CA1 region of the hippocampus. The morphological test showed an increase in the total dendritic length, but a decrease in the number of mature spines in the CA1 region. GFAP labeling increased in the CA1 region and MT-III increased in the CA1 and CA3 regions. Finally, we found a decrease in Zn concentration in the hippocampus after AMPH treatment. An increase in the dopaminergic tone caused by AMPH sensitization generates oxidative stress, neuronal death, and morphological changes in the hippocampus that affect cognitive behaviors like short- and long-term memories.
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Cao DN, Shi JJ, Hao W, Wu N, Li J. Advances and challenges in pharmacotherapeutics for amphetamine-type stimulants addiction. Eur J Pharm. 2016;780:129–35.
Cadet JL, Bisagno V, Milroy CM. Neuropathology of substance use disorders. Acta Neuropathol. 2014;127:91–107.
Sulzer D, Sonders MS, Poulsen NW, Galli A. Mechanisms of neurotransmitter release by amphetamines: a review. Prog Neurobiol. 2005;75:406–33.
United Nations. World Drug Report 2016. New York: United Nations press; 2016.
Robinson TE, Kolb B. Persistent structural modifications in nucleus accumbens and prefrontal cortex neurons produced by previous experience with amphetamine. J Neurosci. 1997;17:8491–7.
Robinson TE, Berridge KC. The neural basis of drug craving: an incentive-sensitization theory of addiction. Brain Res Brain Res Rev. 1993;18:247–91.
Vanderschuren LJ, Pierce RC. Sensitization processes in drug addiction. Curr Top Behav Neurosci. 2010;3:179–95.
Robinson TE, Berridge KC. Review. The incentive sensitization theory of addiction: some current issues. Philos Trans R Soc Lond B Biol Sci. 2008;363:3137–46.
Robinson TE, Berridge KC. Addiction. Annu Rev Psychol. 2003;54:25–53.
Hedou G, Homberg J, Feldon J, Heidbreder CA. Expression of sensitization to amphetamine and dynamics of dopamine neurotransmission in different laminae of the rat medial prefrontal cortex. Neuropharmacology. 2001;40:366–82.
Andersen P, Morris R, Amaral DG, Bliss T, O´Keefe J. The hippocampus book. New York: Oxford University Press; 2007.
Volkow ND, Wang GJ, Fowler JS, Tomasi D. Addiction circuitry in the human brain. Annu Rev Pharm Toxicol. 2012;52:321–36.
Broadbent NJ, Gaskin S, Squire LR, Clark RE. Object recognition memory and the rodent hippocampus. Learn Mem. 2010;17:5–11.
Rosen ZB, Cheung S, Siegelbaum SA. Midbrain dopamine neurons bidirectionally regulate CA3-CA1 synaptic drive. Nat Neurosci. 2015;18:1763–71.
McNamara CG, Dupret D. Two sources of dopamine for the hippocampus. Trends Neurosc. 2017;40:383–4.
Scofield MD, Heinsbroek JA, Gipson CD, Kupchik YM, Spencer S, Smith ACW, et al. The nucleus accumbens: mechanisms of addiction across drug classes reflect the importance of glutamate homeostasis. Pharm Rev. 2016;68:816–71.
Arroyo-García LE, Vázquez-Roque RA, Díaz A, Treviño S, De La Cruz F, Flores G, et al. The effects of non-selective dopamine receptor activation by apomorphine in the mouse hippocampus. Mol Neurobiol. 2018;55:8625–36.
Arroyo-Garcia LE, Rodríguez-Moreno A, Flores G. Apomorphine effects on the hippocampus. Neural Regeneration Res. 2018;13:2064–6.
Cadet JL, Krasnova IN, Jayanthi S, Lyles J. Neurotoxicity of substituted amphetamines: molecular and cellular mechanisms. Neurotox Res. 2007;11:183–202.
Eibl JK, Abdallah Z, Ross GM. Zinc-metallothionein: a potential mediator of antioxidant defence mechanisms in response to dopamine induced stress. Can J Physiol Pharm. 2010;88:305–12.
Cuajungco MP, Lees GJ. Zinc metabolism in the brain: relevance to human neurodegenerative disorders. Neurobiol Dis. 1997;4:137–69.
Wigner P, Czarny P, Galecki P, Su KP, Sliwinski T. The molecular aspects of oxidative & nitrosative stress and the tryptophan catabolites pathway (TRYCATs) as potential causes of depression. Psychiatry Res. 2018;262:566–74.
Hayashi Y, Majewska AK. Dendritic spine geometry: functional implication and regulation. Neuron. 2005;46:529–32.
Segal M. Dendritic spines, synaptic plasticity and neuronal survival: activity shapes dendritic spines to enhance neuronal viability. Eur J Neurosci. 2010;31:2178–84.
Pérez-Rodríguez M, Arroyo-García LE, Prius-Mengual J, Andrade-Talavera Y, Armengol JA, Pérez-Villegas E, et al. Adenosine receptor-mediated developmental loss of spike timing-dependent depression in the hippocampus. Cereb Cortex. 2019;29:3266–3281.
Fiala JC, Spacek J, Harris KM. Dendritic spine pathology: cause or consequence of neurological disorders? Brain Res Brain Res Rev. 2002;39:29–54.
Blanpied TA, Ehlers MD. Microanatomy of dendritic spines: emerging principles of synaptic pathology in psychiatric and neurological disease. Biol Psychiatry. 2004;55:1121–7.
Tendilla-Beltrán H, Antonio Vázquez-Roque R, Judith Vázquez-Hernández A, Garcés-Ramírez L, Flores G. Exploring the dendritic spine pathology in a schizophrenia-related neurodevelopmental animal model. Neuroscience. 2019;396:36–45.
Hernandez-Hernandez EM, Caporal Hernández K, Vázquez-Roque RA, Díaz A, de la Cruz F, Flores G. The neuropeptide-12 improves recognition memory and neural plasticity of the limbic system in old rats. Synapse. 2018;72:e22036.
Tendilla-Beltrán H, Meneses-Prado S, Vázquez-Roque RA, Tapia-Rodríguez M, Vázquez-Hernández AJ, Coatl-Cuaya H, et al. Risperidone ameliorates prefrontal cortex neural atrophy and oxidative/nitrosative stress in brain and peripheral blood of rats with neonatal ventral hippocampus lesion. J Neurosci. 2019;39:8584–99.
Vázquez‐Roque RA, Ramos B, Tecuatl C, Juárez I, Adame A, de la Cruz, et al. Chronic administration of the neurotrophic agent cerebrolysin ameliorates the behavioral and morphological changes induced by neonatal ventral hippocampus lesion in a rat model of schizophrenia. J Neurosci Res. 2012;90:288–306.
Gibb R, Kolb B. A method for vibratome sectioning of Golgi-Cox stained whole rat brain. J Neurosci Methods. 1998;79:1–4.
Paxinos G, Watson C. The rat brain in stereotaxic coordinates, 2nd edn. New York: Academic Press; 1986.
Kolb B, Forgie M, Gibb R, Gorny G, Rowntree S. Age, experience and the changing brain. Neurosci Biobehav Rev. 1998;22:143–59.
Sholl DA. Dendritic organization in the neurons of the visual and motor cortices of the cat. J Anat. 1953;87:387–406.
Flores G, Alquicer G, Silva-Gómez AB, Zaldivar G, Stewart J, Quirion R, et al. Alterations in dendritic morphology of prefrontal cortical and nucleus accumbens neurons in post-pubertal rats after neonatal excitotoxic lesions of the ventral hippocampus. Neuroscience. 2005;133:463–70.
Martínez-Tellez R, Gómez-Villalobos MJ, Flores G. Alteration in dendritic morphology of cortical neurons in rats with diabetes mellitus induced by streptozocin. Brain Res. 2005;1048:108–15.
Silva-Gómez AB, Bermudez M, Quirion R, Srivastava LK, Picazo O, Flores G. Comparative behavioral changes between male and female postpubertal rats following neonatal excitotoxic lesions of the ventral hippocampus. Brain Res. 2003;973:285–92.
Tendilla-Beltrán H, Arroyo-García LE, Díaz A, Camacho-Abrego I, de la Cruz F, Rodriguez-Moreno A, et al. The effects of amphetamine exposure on juvenile rats on the neuronal morphology of the limbic system at prepubertal, pubertal and postpubertal ages. J Chem Neuroanat. 2016;77:68–77.
Bello-Medina PC, Flores G, Quirarte GL, McGaugh JL, Prado-Alcalá RA. Mushroom spine dynamics in medium spiny neurons of dorsal striatum associated with memory of moderate and intense training. Proc Natl Acad Sci USA 2016;113:E6516–25.
Tellez-Merlo G, Morales-Medina JC, Camacho-Abrego I, Juaréz-Díaz I, Aguilar-Alonso P, de la Cruz F, et al. Prenatal immune challenge induces behavioral deficits, neuronal remodeling, and increases brain nitric oxide and zinc levels in the male rat offspring. Neuroscience. 2019;406:594–605.
Eaton DL, Toal BF. Evaluation of the Cd/hemoglobin affinity assay for the rapid determination of metallothionein in biological tissues. Toxicol Appl Pharm. 1982;66:134–42.
Ohkawa H, Ohishi N, Yagi K. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem. 1979;95:351–8.
Tindell AJ, Berridge KC, Jun Z, Peciña S, Aldridge JW. Ventral pallidal neurons code incentive motivation: amplification by mesolimbic sensitization and amphetamine. Eur J Neurosci. 2005;22:2617–34.
Zhu J, Chen Y, Zhao N, Cao G, Dang Y, Chen T. Distinct roles of dopamine D3 receptors in modulating methamphetamine-induced behavioral sensitization and ultrastructural plasticity in the shell of the nucleus accumbens. J Neurosci Res. 2012;90:895–904.
Reske M, Eidt CA, Delis DC, Paulus MP. Nondependent stimulant users of cocaine and prescription amphetamines show verbal learning and memory deficits. Biol Psychiatry. 2010;68:762–9.
Brown MW, Aggleton JP. Recognition memory: what are the roles of the perirhinal cortex and hippocampus? Nat Rev Neurosci. 2001;2:51–61.
Bevins RA, Besheer J. Object recognition in rats and mice: a one-trial non-matching-to-sample learning task to study ‘recognition memory’. Nat Protoc. 2006;1:1306–11.
Kristiansen SL, Nyengaard JR. Digital stereology in neuropathology. APMIS. 2012;120:327–40.
Kuczenski R, Everall IP, Crews L, Adame A, Grant I, Masliah E. Escalating dose-multiple binge methamphetamine exposure results in degeneration of the neocortex and limbic system in the rat. Exp Neurol 2007;207:42–51.
Crombag HS, Gorny G, Li Y, Kolb B, Robinson TE. Opposite effects of amphetamine self-administration experience on dendritic spines in the medial and orbital prefrontal cortex. Cereb Cortex. 2005;15:341–8.
Singer BF, Tanabe LM, Gorny G, Jake-Matthews C, Li Y, Kolb B, et al. Amphetamine-induced changes in dendritic morphology in rat forebrain correspond to associative drug conditioning rather than nonassociative drug sensitization. Biol Psychiatry. 2009;65:835–40.
Regev L, Baram TZ. Corticotropin releasing factor in neuroplasticity. Front Neuroendocrinol. 2014;35:171–9.
Mirzayans R, Andrais B, Kumar P, Murray D. The growing complexity of cancer cell response to DNA-damaging agents: caspase 3 mediates cell death or survival? Int J Mol Sci. 2016;17:708.
Marreiro DD, Cruz KJ, Morais JB, Beserra JB, Severo JS, de Oliveira AR. Zinc and oxidative stress: current mechanisms. Antioxidants. 2017;6.
Travaglia A, La Mendola D, Magrì A, Pietropaolo A, Nicoletti VG, Grasso G, et al. Zinc(II) interactions with brain-derived neurotrophic factor N-terminal peptide fragments: inorganic features and biological perspectives. Inorg Chem. 2013;52:11075–83.
Szewczyk B. Zinc homeostasis and neurodegenerative disorders. Front Aging Neurosci. 2013;5:33.
Baltaci AK, Yuce K, Mogulkoc R. Zinc metabolism and metallothioneins. Biol Tra Ele Rese. 2017;55:223–33.
Juárez-Rebollar D, Rios C, Nava-Ruíz C, Méndez-Armenta M. Metallothionein in brain disorders. Oxid Med Cell Longev. 2017;2017:5828056.
Vasto S, Mocchegiani E, Malavolta M, Cuppari I, Listi F, Nuzzo D, et al. Zinc and inflammatory/immune response in aging. Ann N Y Acad Sci. 2007;1100:111–22.
Kang K, Lee SW, Han JE, Choi JW, Song MR. The complex morphology of reactive astrocytes controlled by fibroblast growth factor signaling. Glia. 2014;62:1328.
Yang K, Broussard JI, Levine AT, Jenson D, Arenkiel BR, Dani JA. Dopamine receptor activity participates in hippocampal synaptic plasticity associated with novel object recognition. Eur J Neurosci. 2017;45:138–46.
Sjulson L, Peyrache A, Cumpelik A, Cassataro D, Buzsáki G. Cocaine place conditioning strengthens location-specific hippocampal coupling to the nucleus accumbens. Neuron. 2018;98:926–34.e5.
Wang X, Pal R, Chen XW, Limpeanchob N, Kumar KN, Michaelis EK. High intrinsic oxidative stress may underlie selective vulnerability of the hippocampal CA1 region. Brain Res. 2005;140:120–6.
Wilde GJ, Pringle AK, Wright P, Iannotti F. Differential vulnerability of the CA1 and CA3 subfields of the hippocampus to superoxide and hydroxyl radicals in vitro. J Neurochem. 1997;69:883–6.
Cadet JL, Patel R, Jayanthi S. Compulsive methamphetamine taking and abstinence in the presence of adverse consequences: epigenetic and transcriptional consequences in the rat brain. Pharm Biochem Behav. 2019;179:98–108.
LEA-G and HT-B acknowledge CONACYT for the fellowship. EB, PA-A, AD, RAV-R, FDLC, EM, and GF acknowledge the “Sistema Nacional de Investigadores” of Mexico for memberships. Thanks to Miguel Tapia-Rodríguez (Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México) for stereological procedures assistance and to Professor Robert Simpson for editing the English language text.
Funding for this study was provided by grants from the PRODEP (CA-BUAP-120) and the CONACYT grant (No. 252808) to GF and MINECO/FEDER (BFU2012-38208) and the Junta de Andalucía (P11-CVI-7290) to AR-M. None of the funding institutions had any further role in the study design, the collection or interpretation of data, analyses, the writing of the report or the decision to submit the paper for publication.
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Arroyo-García, L.E., Tendilla-Beltrán, H., Vázquez-Roque, R.A. et al. Amphetamine sensitization alters hippocampal neuronal morphology and memory and learning behaviors. Mol Psychiatry (2020). https://doi.org/10.1038/s41380-020-0809-2