Journal of Neural Transmission

, Volume 125, Issue 12, pp 1867–1875 | Cite as

Chronic oral methylphenidate treatment increases microglial activation in rats

  • Emily Carias
  • John Hamilton
  • Lisa S. Robison
  • Foteini Delis
  • Rina Eiden
  • Teresa Quattrin
  • Michael Hadjiargyrou
  • David Komatsu
  • Panayotis K. ThanosEmail author
Psychiatry and Preclinical Psychiatric Studies - Original Article


Methylphenidate (MP) is a widely prescribed psychostimulant used to treat attention deficit hyperactivity disorder. Previously, we established a drinking paradigm to deliver MP to rats at doses that result in pharmacokinetic profiles similar to treated patients. In the present study, adolescent male rats were assigned to one of three groups: control (water), low-dose MP (LD; 4/10 mg/kg), and high dose MP (HD; 30/60 mg/kg). Following 3 months of treatment, half of the rats in each group were euthanized, and the remaining rats received only water throughout a 1-month-long abstinence phase. In vitro autoradiography using [3H] PK 11195 was performed to measure microglial activation. HD MP rats showed increased [3H] PK 11195 binding compared to control rats in several cerebral cortical areas: primary somatosensory cortex including jaw (68.6%), upper lip (80.1%), barrel field (88.9%), and trunk (78%) regions, forelimb sensorimotor area (87.3%), secondary somatosensory cortex (72.5%), motor cortices 1 (73.2%) and 2 (69.3%), insular cortex (59.9%); as well as subcortical regions including the thalamus (62.9%), globus pallidus (79.4%) and substantia nigra (22.7%). Additionally, HD MP rats showed greater binding compared to LD MP rats in the hippocampus (60.6%), thalamus (59.6%), substantia nigra (38.5%), and motor 2 cortex (55.3%). Following abstinence, HD MP rats showed no significant differences compared to water controls; however, LD MP rats showed increased binding in pre-limbic cortex (78.1%) and ventromedial caudate putamen (113.8%). These findings indicate that chronic MP results in widespread microglial activation immediately after treatment and following the cessation of treatment in some brain regions.


Methylphenidate Ritalin Attention deficit hyperactivity disorder Autoradiography Microglia Inflammation 



This research was funded by the New York Research Foundation [Q0942016] and the National Institute of Health [R01HD70888].

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted.


  1. Achterberg EJM, van Kerkhof LWM, Damsteegt R, Trezza V, Vanderschuren LJMJ (2015) Methylphenidate and atomoxetine inhibit social play behavior through prefrontal and subcortical limbic mechanisms in rats. J Neurosci 35(1):161–169. CrossRefPubMedPubMedCentralGoogle Scholar
  2. Andreazza AC, Frey BN, Valvassori SS, Zanotto C, Gomes KM, Comim CM, Cassini C, Stertz L, Ribeiro LC, Quevedo J, Kapczinski F, Berk M, Gonçalves CA (2007) DNA damage in rats after treatment with methylphenidate. Prog Neuropsychopharmacol Biol Psychiatry 31(6):1282–1288. CrossRefPubMedGoogle Scholar
  3. Arria AM, DuPont RL (2010) Nonmedical prescription stimulant use among college students: why we need to do something and what we need to do. J Addict Dis 29(4):417–426. CrossRefPubMedPubMedCentralGoogle Scholar
  4. Arria AM, Caldeira KM, O’Grady KE, Vincent KB, Johnson EP, Wish ED (2008) Nonmedical use of prescription stimulants among college students: associations with attention-deficit-hyperactivity disorder and polydrug use. Pharmacotherapy 28(2):156–169. CrossRefPubMedPubMedCentralGoogle Scholar
  5. Banihabib N, Haghi M, Zare S, Farrokhi F (2014) The effect of oral administration of methylphenidate on hippocampal tissue in adult male rats. Neurosurg Q 26(4):315–318(4). CrossRefGoogle Scholar
  6. Bethancourt JA, Camarena ZZ, Britton GB (2009) Exposure to oral methylphenidate from adolescence through young adulthood produces transient effects on hippocampal-sensitive memory in rats. Behav Brain Res 202(1):50–57. CrossRefPubMedGoogle Scholar
  7. Bogle KE, Smith BH (2009) Illicit methylphenidate use: a review of prevalence, availability, pharmacology, and consequences. Curr Drug Abuse Rev 2(2):157–176CrossRefGoogle Scholar
  8. Butovsky O, Ziv Y, Schwartz A, Landa G, Talpalar AE, Pluchino S, Martino G, Schwartz M (2006) Microglia activated by IL-4 or IFN-gamma differentially induce neurogenesis and oligodendrogenesis from adult stem/progenitor cells. Molecular Cellular Neurosci 31(1):149–160. CrossRefGoogle Scholar
  9. Cadet JL, Jayanthi S, Deng X (2005) Methamphetamine-induced neuronal apoptosis involves the activation of multiple death pathways. Rev Neurotox Res 8(3–4):199–206CrossRefGoogle Scholar
  10. Caprioli D, Jupp B, Hong YT, Sawiak SJ, Ferrari V, Wharton L, Williamson DJ, McNabb C, Berry D, Aigbirhio FI, Robbins TW, Fryer TD, Dalley JW (2015) Dissociable rate-dependent effects of oral methylphenidate on impulsivity and D2/3 receptor availability in the striatum. J Neurosci 35(9):3747–3755. CrossRefPubMedPubMedCentralGoogle Scholar
  11. Cubells JF, Rayport S, Rajendran G, Sulzer D (1994) Methamphetamine neurotoxicity involves vacuolation of endocytic organelles and dopamine-dependent intracellular oxidative stress. J Neurosci 14(4):2260–2271CrossRefGoogle Scholar
  12. Dalley JW, Cardinal RN, Robbins TW (2004) Prefrontal executive and cognitive functions in rodents: neural and neurochemical substrates. Neurosci Biobehav Rev 28(7):771–784. CrossRefPubMedGoogle Scholar
  13. Delis F, Weber A, Thanos PK (2017) Chronic oral methylphenidate intake affects white matter morphology and NMDA receptor density in normal rats. In: 27th meeting of the Hellenic Society for Neuroscience, AthensGoogle Scholar
  14. Drouin C, Page M, Waterhouse B (2006) Methylphenidate enhances noradrenergic transmission and suppresses mid- and long-latency sensory responses in the primary somatosensory cortex of awake rats. J Neurophysiol 96(2):622–632. CrossRefPubMedGoogle Scholar
  15. Drouin C, Wang D, Waterhouse BD (2007) Neurophysiological actions of methylphenidate in the primary somatosensory cortex. Synapse 61(12):985–990. doiCrossRefPubMedGoogle Scholar
  16. Fallah G, Moudi S, Hamidia A, Bijani A (2018) Stimulant use in medical students and residents requires more careful attention. Casp J Intern Med 9(1):87–91. CrossRefGoogle Scholar
  17. Filloux F, Townsend JJ (1993) Pre- and postsynaptic neurotoxic effects of dopamine demonstrated by intrastriatal injection. Exp Neurol 119(1):79–88. CrossRefPubMedGoogle Scholar
  18. Gehrmann J, Matsumoto Y, Kreutzberg GW (1995) Microglia: intrinsic immuneffector cell of the brain. Brain Res Brain Res Rev 20(3):269–287CrossRefGoogle Scholar
  19. Gerasimov MR, Franceschi M, Volkow ND, Gifford A, Gatley SJ, Marsteller D, Molina PE, Dewey SL (2000) Comparison between intraperitoneal and oral methylphenidate administration: a microdialysis and locomotor activity study. J Pharmacol Exp Ther 295(1):51–57PubMedGoogle Scholar
  20. Gomes KM, Petronilho FC, Mantovani M, Garbelotto T, Boeck CR, Dal-Pizzol F, Quevedo J (2008) Antioxidant enzyme activities following acute or chronic methylphenidate treatment in young rats. Neurochem Res 33(6):1024–1027CrossRefGoogle Scholar
  21. Gonçalves J, Baptista S, Martins T, Milhazes N, Borges F, Ribeiro Carlos F, Malva João O, Silva Ana P (2010) Methamphetamine-induced neuroinflammation and neuronal dysfunction in the mice hippocampus: preventive effect of indomethacin. Eur J Neurosci 31(2):315–326. CrossRefPubMedGoogle Scholar
  22. Gray JD, Punsoni M, Tabori NE, Melton JT, Fanslow V, Ward MJ, Zupan B, Menzer D, Rice J, Drake CT, Romeo RD, Brake WG, Torres-Reveron A, Milner TA (2007) Methylphenidate administration to juvenile rats alters brain areas involved in cognition, motivated behaviors, appetite, and stress. J Neurosci 27(27):7196–7207. CrossRefPubMedGoogle Scholar
  23. Greenhill L, Beyer DH, Finkleson J, Shaffer D, Biederman J, Conners CK, Gillberg C, Huss M, Jensen P, Kennedy JL, Klein R, Rapoport J, Sagvolden T, Spencer T, Swanson JM, Volkow N (2002a) Guidelines and algorithms for the use of methylphenidate in children with attention-deficit/hyperactivity disorder. J Atten Disord 6(Suppl 1):S89–S100CrossRefGoogle Scholar
  24. Greenhill LL, Findling RL, Swanson JM, Group AS (2002b) A double-blind, placebo-controlled study of modified-release methylphenidate in children with attention-deficit/hyperactivity disorder. Pediatrics 109(3):E39CrossRefGoogle Scholar
  25. Heyser CJ, Pelletier M, Ferris JS (2004) The effects of methylphenidate on novel object exploration in weanling and periadolescent rats. Ann N Y Acad Sci 1021(1):465–469CrossRefGoogle Scholar
  26. Howes SR, Dalley JW, Morrison CH, Robbins TW, Everitt BJ (2000) Leftward shift in the acquisition of cocaine self-administration in isolation-reared rats: relationship to extracellular levels of dopamine, serotonin and glutamate in the nucleus accumbens and amygdala-striatal FOS expression. Psychopharmacology 151(1):55–63CrossRefGoogle Scholar
  27. Jang H, Boltz D, McClaren J, Pani AK, Smeyne M, Korff A, Webster R, Smeyne RJ (2012) Inflammatory effects of highly pathogenic H5N1 influenza virus infection in the CNS of mice. J Neurosci 32(5):1545–1559. CrossRefPubMedPubMedCentralGoogle Scholar
  28. Komatsu DE, Thanos PK, Mary MN, Janda HA, John CM, Robison L, Ananth M, Swanson JM, Volkow ND, Hadjiargyrou M (2012) Chronic exposure to methylphenidate impairs appendicular bone quality in young rats. Bone 50(6):1214–1222CrossRefGoogle Scholar
  29. Kuczenski R, Segal DS (2001) Locomotor effects of acute and repeated threshold doses of amphetamine and methylphenidate: relative roles of dopamine and norepinephrine. J Pharmacol Exp Ther 296(3):876–883PubMedGoogle Scholar
  30. Lakhan SE, Kirchgessner A (2012) Prescription stimulants in individuals with and without attention deficit hyperactivity disorder: misuse, cognitive impact, and adverse effects. Brain Behav 2(5):661–677. CrossRefPubMedPubMedCentralGoogle Scholar
  31. LeBlanc-Duchin D, Taukulis HK (2009) Chronic oral methylphenidate induces post-treatment impairment in recognition and spatial memory in adult rats. Neurobiol Learn Mem 91(3):218–225. CrossRefPubMedGoogle Scholar
  32. Lee JS, Kim BN, Kang E, Lee DS, Kim YK, Chung JK, Lee MC, Cho SC (2005) Regional cerebral blood flow in children with attention deficit hyperactivity disorder: comparison before and after methylphenidate treatment. Hum Brain Mapp 24(3):157–164. doiCrossRefPubMedGoogle Scholar
  33. Motaghinejad M, Motevalian M, Shabab B (2016) Effects of chronic treatment with methylphenidate on oxidative stress and inflammation in hippocampus of adult rats. Neurosci Lett 619:106–113CrossRefGoogle Scholar
  34. Motaghinejad M, Motevalian M, Abdollahi M, Heidari M, Madjd Z (2017a) Topiramate confers neuroprotection against methylphenidate-induced neurodegeneration in dentate gyrus and CA1 regions of Hippocampus via CREB/BDNF pathway in rats. Neurotox Res 31(3):373–399CrossRefGoogle Scholar
  35. Motaghinejad M, Motevalian M, Shabab B, Fatima S (2017b) Effects of acute doses of methylphenidate on inflammation and oxidative stress in isolated hippocampus and cerebral cortex of adult rats. J Neural Transm 124(1):121–131CrossRefGoogle Scholar
  36. Nakajima K, Kohsaka S (2001) Microglia: activation and their significance in the central nervous system. J Biochem 130(2):169–175CrossRefGoogle Scholar
  37. Nieuwenhuys R (2012) The insular cortex: a review. Prog Brain Res 195:123–163. CrossRefPubMedGoogle Scholar
  38. Pedersen MD, Minuzzi L, Wirenfeldt M, Meldgaard M, Slidsborg C, Cumming P, Finsen B (2006) Up-regulation of PK11195 binding in areas of axonal degeneration coincides with early microglial activation in mouse brain. Eur J Neurosci 24(4):991–1000. CrossRefPubMedGoogle Scholar
  39. Persson M, Brantefjord M, Hansson E, Ronnback L (2005) Lipopolysaccharide increases microglial GLT-1 expression and glutamate uptake capacity in vitro by a mechanism dependent on TNF-alpha. Glia 51(2):111–120. CrossRefPubMedGoogle Scholar
  40. Purves A, Fitzpatrick D (2001) Neuroscience, 2nd edn. Sinauer Associates, SunderlandGoogle Scholar
  41. Raghavendra Rao VL, Dogan A, Bowen KK, Dempsey RJ (2000) Traumatic brain injury leads to increased expression of peripheral-type benzodiazepine receptors, neuronal death, and activation of astrocytes and microglia in rat thalamus. Exp Neurol 161(1):102–114. CrossRefPubMedGoogle Scholar
  42. Robison LS, Ananth M, Hadjiargyrou M, Komatsu DE, Thanos PK (2017a) Chronic oral methylphenidate treatment reversibly increases striatal dopamine transporter and dopamine type 1 receptor binding in rats. J Neural Transm 124(5):655–667CrossRefGoogle Scholar
  43. Robison LS, Michaelos M, Gandhi J, Fricke D, Miao E, Lam C-Y, Mauceri A, Vitale M, Lee J, Paeng S (2017b) Sex differences in the physiological and behavioral effects of chronic oral Methylphenidate treatment in rats. Front Behav Neurosci 11:53CrossRefGoogle Scholar
  44. Sadasivan S, Pond BB, Pani AK, Qu C, Jiao Y, Smeyne RJ (2012) Methylphenidate exposure induces dopamine neuron loss and activation of microglia in the basal ganglia of mice. PLoS One 7(3):e33693CrossRefGoogle Scholar
  45. Scherer EBS, da Cunha MJ, Matté C, Schmitz F, Netto CA, Wyse ATS (2010) Methylphenidate affects memory, brain-derived neurotrophic factor immunocontent and brain acetylcholinesterase activity in the rat. Neurobiol Learn Mem 94(2):247–253. CrossRefPubMedGoogle Scholar
  46. Schwartz BS, Bailey-Davis L, Bandeen-Roche K, Pollak J, Hirsch AG, Nau C, Liu AY, Glass TA (2014) Attention deficit disorder, stimulant use, and childhood body mass index trajectory. Pediatrics 133(4):668–676. CrossRefPubMedPubMedCentralGoogle Scholar
  47. Stephenson DT, Schober DA, Smalstig EB, Mincy RE, Gehlert DR, Clemens JA (1995) Peripheral benzodiazepine receptors are colocalized with activated microglia following transient global forebrain ischemia in the rat. J Neurosci 15(7 Pt 2):5263–5274CrossRefGoogle Scholar
  48. Sulzer D, Sonders MS, Poulsen NW, Galli A (2005) Mechanisms of neurotransmitter release by amphetamines: a review. Prog Neurobiol 75(6):406–433. CrossRefGoogle Scholar
  49. Swanson JM, Volkow ND (2008) Increasing use of stimulants warns of potential abuse. Nature 453(7195):586–586. CrossRefPubMedPubMedCentralGoogle Scholar
  50. Thanos PK, Robison LS, Steier J, Hwang YF, Cooper T, Swanson JM, Komatsu DE, Hadjiargyrou M, Volkow ND (2015) A pharmacokinetic model of oral methylphenidate in the rat and effects on behavior. Pharmacol Biochem Behav 131:143–153. CrossRefPubMedPubMedCentralGoogle Scholar
  51. Thanos PK, Kim R, Delis F, Ananth M, Chachati G, Rocco MJ, Masad I, Muniz JA, Grant SC, Gold MS (2016a) Chronic methamphetamine effects on brain structure and function in rats. PloS One 11(6):e0155457CrossRefGoogle Scholar
  52. Thanos PK, Kim R, Delis F, Ananth M, Chachati G, Rocco MJ, Masad I, Muniz JA, Grant SC, Gold MS, Cadet JL, Volkow ND (2016b) Chronic methamphetamine effects on brain structure and function in rats. PloS One 11(6):e0155457. CrossRefPubMedPubMedCentralGoogle Scholar
  53. Thomas DM, Walker PD, Benjamins JA, Geddes TJ, Kuhn DM (2004) Methamphetamine neurotoxicity in dopamine nerve endings of the striatum is associated with microglial activation. J Pharmacol Exp Therapeutics 311(1):1–7. CrossRefGoogle Scholar
  54. Thor DH, Holloway WR (1983) Play soliciting in juvenile male rats: effects of caffeine, amphetamine and methylphenidate. Pharmacol Biochem Behav 19(4):725–727. CrossRefPubMedGoogle Scholar
  55. Trezza V, Damsteegt R, Vanderschuren LJMJ (2009) Conditioned place preference induced by social play behavior: parametrics, extinction, reinstatement and disruption by methylphenidate. Eur Neuropsychopharmacol 19(9):659–669. CrossRefPubMedPubMedCentralGoogle Scholar
  56. Uddin SM, Robison LS, Fricke D, Chernoff E, Hadjiargyrou M, Thanos PK, Komatsu DE (2018) Methylphenidate regulation of osteoclasts in a dose-and sex-dependent manner adversely affects skeletal mechanical integrity. Sci Rep 8(1):1515CrossRefGoogle Scholar
  57. Vanderschuren LJMJ, Trezza V, Griffioen-Roose S, Schiepers OJG, Van Leeuwen N, De Vries TJ, Schoffelmeer ANM (2008) Methylphenidate disrupts social play behavior in adolescent rats. Neuropsychopharmacology 33:2946. CrossRefPubMedPubMedCentralGoogle Scholar
  58. Venneti S, Lopresti BJ, Wiley CA (2006) The peripheral benzodiazepine receptor in microglia: from pathology to imaging. Prog Neurobiol 80(6):308–322. CrossRefPubMedPubMedCentralGoogle Scholar
  59. Visser SN, Danielson ML, Bitsko RH, Holbrook JR, Kogan MD, Ghandour RM, Perou R, Blumberg SJ (2014) Trends in the parent-report of health care provider-diagnosed and medicated attention-deficit/hyperactivity disorder: United States, 2003–2011. J Am Acad Child Adolesc Psychiatry 53(1):34–46 e32. CrossRefPubMedPubMedCentralGoogle Scholar
  60. Volkow ND, Wang GJ, Fischman MW, Foltin RW, Fowler JS, Abumrad NN, Vitkun S, Logan J, Gatley SJ, Pappas N, Hitzemann R, Shea CE (1997) Relationship between subjective effects of cocaine and dopamine transporter occupancy. Nature 386(6627):827–830. CrossRefPubMedGoogle Scholar
  61. Volkow ND, Wang G-J, Fowler JS, Logan J, Gerasimov M, Maynard L, Ding Y-S, Gatley SJ, Gifford A, Franceschi D (2001) Therapeutic doses of oral methylphenidate significantly increase extracellular dopamine in the human brain. J Neurosci 21(2):RC121CrossRefGoogle Scholar
  62. Volkow ND, Fowler JS, Wang G, Ding Y, Gatley SJ (2002) Mechanism of action of methylphenidate: insights from PET imaging studies. J Attent Disord 6(Suppl 1):S31–S43CrossRefGoogle Scholar
  63. Vowinckel E, Reutens D, Becher B, Verge G, Evans A, Owens T, Antel JP (1997) PK11195 binding to the peripheral benzodiazepine receptor as a marker of microglia activation in multiple sclerosis and experimental autoimmune encephalomyelitis. J Neurosci Res 50 (2):345–353.;2-5 CrossRefPubMedGoogle Scholar
  64. Yang PB, Swann AC, Dafny N (2006) Chronic methylphenidate modulates locomotor activity and sensory evoked responses in the VTA and NAc of freely behaving rats. Neuropharmacology 51(3):546–556. CrossRefPubMedGoogle Scholar
  65. Zhang CL, Feng ZJ, Liu Y, Ji XH, Peng JY, Zhang XH, Zhen XC, Li BM (2012) Methylphenidate enhances NMDA-receptor response in medial prefrontal cortex via sigma-1 receptor: a novel mechanism for methylphenidate action. PloS One 7(12):e51910. CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2018

Authors and Affiliations

  • Emily Carias
    • 1
  • John Hamilton
    • 1
  • Lisa S. Robison
    • 2
  • Foteini Delis
    • 3
  • Rina Eiden
    • 4
  • Teresa Quattrin
    • 5
  • Michael Hadjiargyrou
    • 6
  • David Komatsu
    • 7
  • Panayotis K. Thanos
    • 1
    Email author
  1. 1.Behavioral Neuropharmacology and Neuroimaging Laboratory on Addictions (BNNLA), Research Institute on Addictions, Department of Pharmacology and Toxicology, Jacobs School of Medicine and Biomedical SciencesUniversity at BuffaloBuffaloUSA
  2. 2.Department of Neuroscience and Experimental TherapeuticsAlbany Medical CollegeAlbanyUSA
  3. 3.Department of Pharmacology, Medical SchoolUniversity of IoanninaIoanninaGreece
  4. 4.Department of PsychologyUniversity at BuffaloBuffaloUSA
  5. 5.Women and Children’s Hospital of Buffalo, Department of Pediatrics, School of Medicine and Biomedical SciencesUniversity at BuffaloBuffaloUSA
  6. 6.Department of Life SciencesNew York Institute of TechnologyOld WestburyUSA
  7. 7.Department of OrthopedicsStony Brook UniversityStony BrookUSA

Personalised recommendations