Brain Structure and Function

, Volume 223, Issue 5, pp 2197–2211 | Cite as

Neuroadaptations in the dentate gyrus following contextual cued reinstatement of methamphetamine seeking

  • Yoshio Takashima
  • McKenzie J. Fannon
  • Melissa H. Galinato
  • Noah L. Steiner
  • Michelle An
  • Alice E. Zemljic-Harpf
  • Sucharita S. Somkuwar
  • Brian P. Head
  • Chitra D. MandyamEmail author
Original Article


Abstinence from unregulated methamphetamine self-administration increases hippocampal dependent, context-driven reinstatement of methamphetamine seeking. The current study tested the hypothesis that alterations in the functional properties of granule cell neurons (GCNs) in the dentate gyrus (DG) of the hippocampus in concert with altered expression of synaptic plasticity-related proteins and ultrastructural changes in the DG are associated with enhanced context-driven methamphetamine-seeking behavior. Whole-cell patch-clamp recordings were performed in acute brain slices from methamphetamine naïve (controls) and methamphetamine experienced animals (during acute withdrawal, during abstinence, after extinction and after reinstatement). Spontaneous excitatory postsynaptic currents (sEPSCs) and intrinsic excitability were recorded from GCNs. Reinstatement of methamphetamine seeking increased sEPSC frequency and produced larger amplitude responses in GCNs compared to controls and all other groups. Reinstatement of methamphetamine seeking reduced spiking capability in GCNs compared to controls, and all other groups, as indicated by reduced intrinsic spiking elicited by increasing current injections, membrane resistance and fast after hyperpolarization. In rats that reinstated methamphetamine seeking, these altered electrophysiological properties of GCNs were associated with enhanced expression of Fos, GluN2A subunits and PSD95 and reduced expression of GABAA subunits in the DG and enhanced expression of synaptic PSD in the molecular layer. The alterations in functional properties of GCNs and plasticity related proteins in the DG paralleled with no changes in structure of microglial cells in the DG. Taken together, our results demonstrate that enhanced reinstatement of methamphetamine seeking results in alterations in intrinsic spiking and spontaneous glutamatergic synaptic transmission in the GCNs and concomitant increases in neuronal activation of GCNs, and expression of GluNs and decreases in GABAA subunits that may contribute to the altered synaptic connectivity—neuronal circuitry—and activity in the hippocampus, and enhance propensity for relapse.


Granule cell neurons Whole-cell patch-clamp recording Fos GluN Electron microscopy Microglia 



This work was supported by grants from the National Science Foundation, USA (DGE-1144086 to M.H.G), National Institute on Drug Abuse, USA (DA034140 to C.D.M.), National Institute of Alcoholism and Alcohol Abuse, USA (AA020098, AA06420 to C.D.M.) and start-up funds from the Veterans Medical Research Foundation to C.D.M. We sincerely thank Dr. Bryan W. Luikart from the Geisel School of Medicine, Dartmouth College, for his scientific collaboration and extensive discussion on the electrophysiology data. We thank Ying Jones at the UCSD EM core for preparation of the brain samples and training on the EM. We thank Ryan Ostrom from the division of biological sciences independent study program at UCSD and Jasmine Guevara from the NIDA summer internship program for assistance with animal behavior.

Compliance with ethical standards

Conflict of interest

The authors declare no competing financial interests in relation to the work described.


  1. Ali DW, Salter MW (2001) NMDA receptor regulation by Src kinase signalling in excitatory synaptic transmission and plasticity. Curr Opin Neurobiol 11:336–342CrossRefPubMedGoogle Scholar
  2. Amaral DG, Witter MP (1989) The three-dimensional organization of the hippocampal formation: a review of anatomical data. Neuroscience 31:571–591CrossRefPubMedGoogle Scholar
  3. Andersen P, Holmqvist B, Voorhoeve PE (1966a) Entorhinal activation of dentate granule cells. Acta Physiol Scand 66:448–460CrossRefPubMedGoogle Scholar
  4. Andersen P, Holmqvist B, Voorhoeve PE (1966b) Excitatory synapses on hippocampal apical dendrites activated by entorhinal stimulation. Acta Physiol Scand 66:461–472CrossRefPubMedGoogle Scholar
  5. Andersen P, Bliss TV, Skrede KK (1971) Unit analysis of hippocampal polulation spikes. Exp Brain Res 13:208–221PubMedGoogle Scholar
  6. Benedetti BL, Takashima Y, Wen JA, Urban-Ciecko J, Barth AL (2013) Differential wiring of layer 2/3 neurons drives sparse and reliable firing during neocortical development. Cereb Cortex 23:2690–2699CrossRefPubMedGoogle Scholar
  7. Brenner R, Chen QH, Vilaythong A, Toney GM, Noebels JL, Aldrich RW (2005) BK channel beta4 subunit reduces dentate gyrus excitability and protects against temporal lobe seizures. Nat Neurosci 8:1752–1759CrossRefPubMedGoogle Scholar
  8. Bu Q, Lv L, Yan G, Deng P, Wang Y, Zhou J, Yang Y, Li Y, Cen X (2013) NMR-based metabonomic in hippocampus, nucleus accumbens and prefrontal cortex of methamphetamine-sensitized rats. Neurotoxicology 36:17–23CrossRefPubMedGoogle Scholar
  9. Buchanan JB, Sparkman NL, Johnson RW (2010) A neurotoxic regimen of methamphetamine exacerbates the febrile and neuroinflammatory response to a subsequent peripheral immune stimulus. J Neuroinflamm 7:82CrossRefGoogle Scholar
  10. Burgess N, Maguire EA, O’Keefe J (2002) The human hippocampus and spatial and episodic memory. Neuron 35:625–641CrossRefPubMedGoogle Scholar
  11. Coulter DA, Carlson GC (2007) Functional regulation of the dentate gyrus by GABA-mediated inhibition. Prog Brain Res 163:235–243CrossRefPubMedGoogle Scholar
  12. Criado JR, Gombart LM, Huitron-Resendiz S, Henriksen SJ (2000) Neuroadaptations in dentate gyrus function following repeated methamphetamine administration. Synapse 37:163–166CrossRefPubMedGoogle Scholar
  13. Crombag HS, Gorny G, Li Y, Kolb B, Robinson TE (2005) Opposite effects of amphetamine self-administration experience on dendritic spines in the medial and orbital prefrontal cortex. Cereb Cortex 15:341–348CrossRefPubMedGoogle Scholar
  14. Daumann J, Koester P, Becker B, Wagner D, Imperati D, Gouzoulis-Mayfrank E, Tittgemeyer M (2011) Medial prefrontal gray matter volume reductions in users of amphetamine-type stimulants revealed by combined tract-based spatial statistics and voxel-based morphometry. Neuroimage 54:794–801CrossRefPubMedGoogle Scholar
  15. De Koninck Y, Mody I (1994) Noise analysis of miniature IPSCs in adult rat brain slices: properties and modulation of synaptic GABAA receptor channels. J Neurophysiol 71:1318–1335CrossRefPubMedGoogle Scholar
  16. Fuchs RA, Evans KA, Ledford CC, Parker MP, Case JM, Mehta RH, See RE (2005) The role of the dorsomedial prefrontal cortex, basolateral amygdala, and dorsal hippocampus in contextual reinstatement of cocaine seeking in rats. Neuropsychopharmacology 30:296–309CrossRefPubMedGoogle Scholar
  17. Galinato MH, Orio L, Mandyam CD (2015) Methamphetamine differentially affects BDNF and cell death factors in anatomically defined regions of the hippocampus. Neuroscience 286:97–108CrossRefPubMedGoogle Scholar
  18. Galinato MH, Lockner JW, Fannon-Pavlich MJ, Sobieraj JC, Staples MC, Somkuwar SS, Ghofranian A, Chaing S, Navarro AI, Joea A, Luikart BW, Janda KD, Heyser C, Koob GF, Mandyam CD (2017) A synthetic small-molecule Isoxazole-9 protects against methamphetamine relapse. Mol Psychiatry.
  19. Glykys J, Mody I (2006) Hippocampal network hyperactivity after selective reduction of tonic inhibition in GABA A receptor alpha5 subunit-deficient mice. J Neurophysiol 95:2796–2807CrossRefPubMedGoogle Scholar
  20. Glykys J, Mody I (2007) The main source of ambient GABA responsible for tonic inhibition in the mouse hippocampus. J Physiol 582:1163–1178CrossRefPubMedPubMedCentralGoogle Scholar
  21. Han W, Wang F, Qi J, Wang F, Zhang L, Zhao S, Song M, Wu C, Yang J (2012) NMDA receptors in the medial prefrontal cortex and the dorsal hippocampus regulate methamphetamine-induced hyperactivity and extracellular amino acid release in mice. Behav Brain Res 232:44–52CrossRefPubMedGoogle Scholar
  22. Herd MB, Haythornthwaite AR, Rosahl TW, Wafford KA, Homanics GE, Lambert JJ, Belelli D (2008) The expression of GABAA beta subunit isoforms in synaptic and extrasynaptic receptor populations of mouse dentate gyrus granule cells. J Physiol 586:989–1004CrossRefPubMedGoogle Scholar
  23. Hiranita T, Nawata Y, Sakimura K, Anggadiredja K, Yamamoto T (2006) Suppression of methamphetamine-seeking behavior by nicotinic agonists. Proc Natl Acad Sci USA 103:8523–8527CrossRefPubMedPubMedCentralGoogle Scholar
  24. Jung MW, McNaughton BL (1993) Spatial selectivity of unit activity in the hippocampal granular layer. Hippocampus 3:165–182CrossRefPubMedGoogle Scholar
  25. Kim YT, Lee JJ, Song HJ, Kim JH, Kwon DH, Kim MN, Yoo DS, Lee HJ, Kim HJ, Chang Y (2010) Alterations in cortical activity of male methamphetamine abusers performing an empathy task: fMRI study. Hum Psychopharmacol 25:63–70CrossRefPubMedGoogle Scholar
  26. Kim A, Zamora-Martinez ER, Edwards S, Mandyam CD (2015) Structural reorganization of pyramidal neurons in the medial prefrontal cortex of alcohol dependent rats is associated with altered glial plasticity. Brain Struct Funct 220:1705–1720CrossRefPubMedGoogle Scholar
  27. Lavezzari G, McCallum J, Lee R, Roche KW (2003) Differential binding of the AP-2 adaptor complex and PSD-95 to the C-terminus of the NMDA receptor subunit NR2B regulates surface expression. Neuropharmacology 45:729–737CrossRefPubMedGoogle Scholar
  28. Licznerski P, Duman RS (2013) Remodeling of axo-spinous synapses in the pathophysiology and treatment of depression. Neuroscience 251:33–50CrossRefPubMedGoogle Scholar
  29. Lin M, Sambo D, Khoshbouei H (2016) Methamphetamine regulation of firing activity of dopamine neurons. J Neurosci 36:10376–10391CrossRefPubMedPubMedCentralGoogle Scholar
  30. Liu YB, Lio PA, Pasternak JF, Trommer BL (1996) Developmental changes in membrane properties and postsynaptic currents of granule cells in rat dentate gyrus. J Neurophysiol 76:1074–1088CrossRefPubMedGoogle Scholar
  31. Liu X, Tilwalli S, Ye G, Lio PA, Pasternak JF, Trommer BL (2000) Morphologic and electrophysiologic maturation in developing dentate gyrus granule cells. Brain Res 856:202–212CrossRefPubMedGoogle Scholar
  32. Mandyam CD, Norris RD, Eisch AJ (2004) Chronic morphine induces premature mitosis of proliferating cells in the adult mouse subgranular zone. J Neurosci Res 76:783–794CrossRefPubMedGoogle Scholar
  33. Mandyam CD, Wee S, Crawford EF, Eisch AJ, Richardson HN, Koob GF (2008) Varied access to intravenous methamphetamine self-administration differentially alters adult hippocampal neurogenesis. Biol Psychiatry 64:958–965CrossRefPubMedPubMedCentralGoogle Scholar
  34. Mehranfard N, Gholamipour-Badie H, Motamedi F, Janahmadi M, Naderi N (2015) Long-term increases in BK potassium channel underlie increased action potential firing in dentate granule neurons following pilocarpine-induced status epilepticus in rats. Neurosci Lett 585:88–91CrossRefPubMedGoogle Scholar
  35. Meyers RA, Zavala AR, Speer CM, Neisewander JL (2006) Dorsal hippocampus inhibition disrupts acquisition and expression, but not consolidation, of cocaine conditioned place preference. Behav Neurosci 120:401–412CrossRefPubMedGoogle Scholar
  36. Morales AM, Lee B, Hellemann G, O’Neill J, London ED (2012) Gray-matter volume in methamphetamine dependence: cigarette smoking and changes with abstinence from methamphetamine. Drug Alcohol Depend 125:230–238CrossRefPubMedPubMedCentralGoogle Scholar
  37. Morris P, Bachelard H (2003) Reflections on the application of 13C-MRS to research on brain metabolism. NMR in biomedicine 16:303–312CrossRefPubMedGoogle Scholar
  38. Nakama H, Chang L, Fein G, Shimotsu R, Jiang CS, Ernst T (2011) Methamphetamine users show greater than normal age-related cortical gray matter loss. Addiction 106:1474–1483CrossRefPubMedPubMedCentralGoogle Scholar
  39. Nakazawa T, Komai S, Tezuka T, Hisatsune C, Umemori H, Semba K, Mishina M, Manabe T, Yamamoto T (2001) Characterization of fyn-mediated tyrosine phosphorylation sites on GluR epsilon 2 (NR2B) subunit of the N-methyl-d-aspartate receptor. J Biol Chem 276:693–699CrossRefPubMedGoogle Scholar
  40. Nakazawa T, Komai S, Watabe AM, Kiyama Y, Fukaya M, Arima-Yoshida F, Horai R, Sudo K, Ebine K, Delawary M, Goto J, Umemori H, Tezuka T, Iwakura Y, Watanabe M, Yamamoto T, Manabe T (2006) NR2B tyrosine phosphorylation modulates fear learning as well as amygdaloid synaptic plasticity. Embo j 25:2867–2877CrossRefPubMedPubMedCentralGoogle Scholar
  41. Nikonenko I, Jourdain P, Alberi S, Toni N, Muller D (2002) Activity-induced changes of spine morphology. Hippocampus 12:585–591CrossRefPubMedGoogle Scholar
  42. Orikabe L, Yamasue H, Inoue H, Takayanagi Y, Mozue Y, Sudo Y, Ishii T, Itokawa M, Suzuki M, Kurachi M, Okazaki Y, Kasai K (2011) Reduced amygdala and hippocampal volumes in patients with methamphetamine psychosis. Schizophr Res 132:183–189CrossRefPubMedGoogle Scholar
  43. Price KL, DeSantis SM, Simpson AN, Tolliver BK, McRae-Clark AL, Saladin ME, Baker NL, Wagner MT, Brady KT (2011) The impact of clinical and demographic variables on cognitive performance in methamphetamine-dependent individuals in rural South Carolina. Am J Addict 20:447–455CrossRefPubMedPubMedCentralGoogle Scholar
  44. Prybylowski K, Chang K, Sans N, Kan L, Vicini S, Wenthold RJ (2005) The synaptic localization of NR2B-containing NMDA receptors is controlled by interactions with PDZ proteins and AP-2. Neuron 47:845–857CrossRefPubMedPubMedCentralGoogle Scholar
  45. Recinto P, Samant AR, Chavez G, Kim A, Yuan CJ, Soleiman M, Grant Y, Edwards S, Wee S, Koob GF, George O, Mandyam CD (2012) Levels of neural progenitors in the hippocampus predict memory impairment and relapse to drug seeking as a function of excessive methamphetamine self-administration. Neuropsychopharmacology 37:1275–1287CrossRefPubMedGoogle Scholar
  46. SAMHSA (2008) Results from the 2007 National Survey on Drug Use and Health: Detailed Tables. Substance Abuse and Mental Health Services Administration, Office of Applied StudiesGoogle Scholar
  47. Scharfman HE, Schwartzkroin PA (1989) Protection of dentate hilar cells from prolonged stimulation by intracellular calcium chelation. Science 246:257–260CrossRefPubMedGoogle Scholar
  48. Schwartz DL, Mitchell AD, Lahna DL, Luber HS, Huckans MS, Mitchell SH, Hoffman WF (2010) Global and local morphometric differences in recently abstinent methamphetamine-dependent individuals. Neuroimage 50:1392–1401CrossRefPubMedPubMedCentralGoogle Scholar
  49. Shaham Y, Shalev U, Lu L, De Wit H, Stewart J (2003) The reinstatement model of drug relapse: history, methodology and major findings. Psychopharmacology 168:3–20CrossRefPubMedGoogle Scholar
  50. Shinohara Y, Hirase H (2009) Size and receptor density of glutamatergic synapses: a viewpoint from left-right asymmetry of CA3–CA1 connections. Front Neuroanat 3:10CrossRefPubMedPubMedCentralGoogle Scholar
  51. Shipton OA, Paulsen O (2014) GluN2A and GluN2B subunit-containing NMDA receptors in hippocampal plasticity. Philos Trans R Soc Lond B Biol Sci 369:20130163CrossRefPubMedPubMedCentralGoogle Scholar
  52. Sobieraj JC, Kim A, Fannon MJ, Mandyam CD (2016) Chronic wheel running-induced reduction of extinction and reinstatement of methamphetamine seeking in methamphetamine dependent rats is associated with reduced number of periaqueductal gray dopamine neurons. Brain Struct Funct 221(1):261–276CrossRefPubMedGoogle Scholar
  53. Somkuwar SS, Fannon-Pavlich MJ, Ghofranian A, Quigley JA, Dutta RR, Galinato MH, Mandyam CD (2016) Wheel running reduces ethanol seeking by increasing neuronal activation and reducing oligodendroglial/neuroinflammatory factors in the medial prefrontal cortex. Brain Behav Immun 58:357–368CrossRefPubMedPubMedCentralGoogle Scholar
  54. Spruston N, Jonas P, Sakmann B (1995) Dendritic glutamate receptor channels in rat hippocampal CA3 and CA1 pyramidal neurons. J Physiol 482(Pt 2):325–352CrossRefPubMedPubMedCentralGoogle Scholar
  55. Squire LR, Stark CE, Clark RE (2004) The medial temporal lobe. Annu Rev Neurosci 27:279–306CrossRefPubMedGoogle Scholar
  56. Stell BM, Brickley SG, Tang CY, Farrant M, Mody I (2003) Neuroactive steroids reduce neuronal excitability by selectively enhancing tonic inhibition mediated by delta subunit-containing GABAA receptors. Proc Natl Acad Sci USA 100:14439–14444CrossRefPubMedPubMedCentralGoogle Scholar
  57. Swanson LW, Wyss JM, Cowan WM (1978) An autoradiographic study of the organization of intrahippocampal association pathways in the rat. J Comp Neurol 181:681–715CrossRefPubMedGoogle Scholar
  58. Swant J, Chirwa S, Stanwood G, Khoshbouei H (2010) Methamphetamine reduces LTP and increases baseline synaptic transmission in the CA1 region of mouse hippocampus. PLoS One 5:e11382CrossRefPubMedPubMedCentralGoogle Scholar
  59. Thompson PM, Hayashi KM, Simon SL, Geaga JA, Hong MS, Sui Y, Lee JY, Toga AW, Ling W, London ED (2004) Structural abnormalities in the brains of human subjects who use methamphetamine. J Neurosci 24:6028–6036CrossRefPubMedGoogle Scholar
  60. Wang B, Bugay V, Ling L, Chuang HH, Jaffe DB, Brenner R (2016) Knockout of the BK beta4-subunit promotes a functional coupling of BK channels and ryanodine receptors that mediate a fAHP-induced increase in excitability. J Neurophysiol 116:456–465CrossRefPubMedPubMedCentralGoogle Scholar
  61. Weber E, Blackstone K, Iudicello JE, Morgan EE, Grant I, Moore DJ, Woods SP (2012) Neurocognitive deficits are associated with unemployment in chronic methamphetamine users. Drug Alcohol Depend 125:146–153CrossRefPubMedPubMedCentralGoogle Scholar
  62. Wells AM, Lasseter HC, Xie X, Cowhey KE, Reittinger AM, Fuchs RA (2011) Interaction between the basolateral amygdala and dorsal hippocampus is critical for cocaine memory reconsolidation and subsequent drug context-induced cocaine-seeking behavior in rats. Learn Mem 18:693–702CrossRefPubMedPubMedCentralGoogle Scholar
  63. Wells AM, Xie X, Higginbotham JA, Arguello AA, Healey KL, Blanton M, Fuchs RA (2016) Contribution of an SFK-mediated signaling pathway in the dorsal hippocampus to cocaine-memory reconsolidation in rats. Neuropsychopharmacology 41:675–685CrossRefPubMedGoogle Scholar
  64. Wheal HV, Chen Y, Mitchell J, Schachner M, Maerz W, Wieland H, Van Rossum D, Kirsch J (1998) Molecular mechanisms that underlie structural and functional changes at the postsynaptic membrane during synaptic plasticity. Prog Neurobiol 55:611–640CrossRefPubMedGoogle Scholar
  65. Yassin L, Benedetti BL, Jouhanneau JS, Wen JA, Poulet JF, Barth AL (2010) An embedded subnetwork of highly active neurons in the neocortex. Neuron 68:1043–1050CrossRefPubMedPubMedCentralGoogle Scholar
  66. Yeckel MF, Berger TW (1990) Feedforward excitation of the hippocampus by afferents from the entorhinal cortex: redefinition of the role of the trisynaptic pathway. Proc Natl Acad Sci USA 87:5832–5836CrossRefPubMedPubMedCentralGoogle Scholar
  67. Yuan CJ, Quiocho JM, Kim A, Wee S, Mandyam CD (2011) Extended access methamphetamine decreases immature neurons in the hippocampus which results from loss and altered development of neural progenitors without altered dynamics of the S-phase of the cell cycle. Pharmacol Biochem Behav 100:98–108CrossRefPubMedPubMedCentralGoogle Scholar
  68. Zhao C, Teng EM, Summers RG Jr, Ming GL, Gage FH (2006) Distinct morphological stages of dentate granule neuron maturation in the adult mouse hippocampus. J Neurosci 26:3–11CrossRefPubMedGoogle Scholar
  69. Zilberter Y, Kaiser KM, Sakmann B (1999) Dendritic GABA release depresses excitatory transmission between layer 2/3 pyramidal and bitufted neurons in rat neocortex. Neuron 24:979–988CrossRefPubMedGoogle Scholar
  70. Zou JY, Crews FT (2005) TNF alpha potentiates glutamate neurotoxicity by inhibiting glutamate uptake in organotypic brain slice cultures: neuroprotection by NF kappa B inhibition. Brain Res 1034:11–24CrossRefPubMedGoogle Scholar

Copyright information

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

Authors and Affiliations

  • Yoshio Takashima
    • 1
    • 2
  • McKenzie J. Fannon
    • 3
  • Melissa H. Galinato
    • 1
    • 3
  • Noah L. Steiner
    • 3
  • Michelle An
    • 3
  • Alice E. Zemljic-Harpf
    • 2
    • 3
  • Sucharita S. Somkuwar
    • 3
  • Brian P. Head
    • 2
    • 3
  • Chitra D. Mandyam
    • 1
    • 2
    • 3
    Email author
  1. 1.Department of NeuroscienceUniversity of California San DiegoSan DiegoUSA
  2. 2.Department of AnesthesiologyUniversity of California San DiegoSan DiegoUSA
  3. 3.VA San Diego Healthcare SystemSan DiegoUSA

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