Naunyn-Schmiedeberg's Archives of Pharmacology

, Volume 391, Issue 4, pp 423–434 | Cite as

Methamphetamine withdrawal induces activation of CRF neurons in the brain stress system in parallel with an increased activity of cardiac sympathetic pathways

  • Juan Antonio García-Carmona
  • Polymnia Georgiou
  • Panos Zanos
  • Alexis Bailey
  • Maria Luisa Laorden
Original Article


Methamphetamine (METH) addiction is a major public health problem in some countries. There is evidence to suggest that METH use is associated with increased risk of developing cardiovascular problems. Here, we investigated the effects of chronic METH administration and withdrawal on the activation of the brain stress system and cardiac sympathetic pathways. Mice were treated with METH (2 mg/kg, i.p.) for 10 days and left to spontaneous withdraw for 7 days. The number of corticotrophin-releasing factor (CRF), c-Fos, and CRF/c-Fos neurons was measured by immunohistochemistry in the paraventricular nucleus of the hypothalamus (PVN) and the oval region of the bed nucleus of stria terminalis (ovBNST), two regions associated with cardiac sympathetic control. In parallel, levels of catechol-o-methyl-transferase (COMT), tyrosine hydroxylase (TH), and heat shock protein 27 (Hsp27) were measured in the heart. In the brain, chronic-METH treatment enhanced the number of c-Fos neurons and the CRF neurons with c-Fos signal (CRF+/c-Fos+) in PVN and ovBNST. METH withdrawal increased the number of CRF+ neurons. In the heart, METH administration induced an increase in soluble (S)-COMT and membrane-bound (MB)-COMT without changes in phospho (p)-TH, Hsp27, or pHsp27. Similarly, METH withdrawal increased the expression of S- and MB-COMT. In contrast to chronic treatment, METH withdrawal enhanced levels of (p)TH and (p)Hsp27 in the heart. Overall, our results demonstrate that chronic METH administration and withdrawal activate the brain CRF systems associated with the heart sympathetic control and point towards a METH withdrawal induced activation of sympathetic pathways in the heart. Our findings provide further insight in the mechanism underlining the cardiovascular risk associated with METH use and proposes targets for its treatment.


Methamphetamine Addiction Withdrawal CRF COMT Hsp27 Heart 



The authors want to thank Dr. Julie Howarth and Ms. Ashleigh Thompson for her assistance with cardiac histopathology.

Funding information

Funding for this study was provided by a Royal Society grant (RG120556; P.I. Alexis Bailey). The sponsors had no involvement in the design of the study and in the collection, analyses, and interpretation of the data nor in the writing of the manuscript and the decision to submit this article for publication.

Compliance with ethical standards

All experimental procedures were conducted in accordance with the UK Animal Scientific Procedures Act (1986).


  1. Abekawa T, Ohmori T, Koyama T (1994) Effects of repeated administration of a high dose of methamphetamine on dopamine and glutamate release in rat striatum and nucleus accumbens. Brain Res 643(1–2):276–281. CrossRefPubMedGoogle Scholar
  2. Ammon-Treiber S, Grecksch G, Stumm R, Riechert U, Tischmeyer H, Reichenauer A, Höllt V (2004) Rapid, transient, and dose-dependent expression of hsp70 messenger RNA in the rat brain after morphine treatment. Cell Stress Chaperones 9(2):182–197. CrossRefPubMedPubMedCentralGoogle Scholar
  3. Beuten J, Payne TJ, Ma JZ, Li MD (2005) Significant association of catechol-O-methyl transferase (COMT) haplotypes with nicotine dependence in male and female smokers of two ethnic populations. Neuropsychopharmacology 31:675–684CrossRefGoogle Scholar
  4. Chu CP, Qiu DL, Kato K, Kunitake T, Watanabe S, Yu NS, Nakazato M, Kannan H (2004) Central stresscopin modulates cardiovascular function through the adrenal medulla in conscious rats. Regul Pept 119(1-2):53–59. CrossRefPubMedGoogle Scholar
  5. Ciccarone D (2011) Stimulant abuse: pharmacology, cocaine, methamphetamine, treatment, attempts at pharmacotherapy. Primary care 38(1):41–58, v-vi. CrossRefPubMedPubMedCentralGoogle Scholar
  6. 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):2260–2271PubMedGoogle Scholar
  7. Daniel SE, Rainnie DG (2016) Stress modulation of opposing circuits in the bed nucleus of the stria terminalis. Neuropsychopharmacology 41(1):103–125CrossRefPubMedGoogle Scholar
  8. Deng X, Feng X, Li S, Gao Y, Yu B, Li G (2015) Influence of the hypothalamic paraventricular nucleus (PVN) on heart rate variability (HRV) in rat hearts via electronic lesion. Biomed Mater Eng 26(Suppl 1):S487–S495. PubMedGoogle Scholar
  9. Dettmeyer R, Friedrich K, Schmidt P, Madea B (2009) Heroin-associated myocardial damages—conventional and immunohistochemical investigations. Forensic Sci Int 187(1–3):42–46. CrossRefPubMedGoogle Scholar
  10. Dietrich JB, Arpin-Bott MP, Kao D, Dirrig-Grosch S, Aunis D, Zwiller J (2007) Cocaine induces the expression of homer 1b/c, homer 3a/b, and hsp 27 proteins in rat cerebellum. Synapse 61(8):587–594. CrossRefPubMedGoogle Scholar
  11. Dunkley PR, Bobrovskaya L, Graham ME, von Nagy-Felsobuki EI, Dickson PW (2004) Tyrosine hydroxylase phosphorylation: regulation and consequences. J Neurochem 91(5):1025–1043. CrossRefPubMedGoogle Scholar
  12. Ersche KD, Roiser JP, Lucas M, Domenici E, Robbins TW, Bullmore ET (2011) Peripheral biomarkers of cognitive response to dopamine receptor agonist treatment. Psychopharmacology 214(4):779–789. CrossRefPubMedGoogle Scholar
  13. Francesconi W, Berton F, Repunte-Canonigo V, Hagihara K, Thurbon D, Lekic D, Specio SE, Greenwell TN, Chen SA, Rice KC, Richardson HN, O’Dell LE, Zorrilla EP, Morales M, Koob GF, Sanna PP (2009) Protracted withdrawal from alcohol and drugs of abuse impairs long-term potentiation of intrinsic excitability in the juxtacapsular bed nucleus of the stria terminalis. J Neurosci 29(17):5389–5401. CrossRefPubMedPubMedCentralGoogle Scholar
  14. Garcia-Carmona JA, Milanes MV, Laorden ML (2013) Brain stress system response after morphine-conditioned place preference. Int J Neuropsychopharmacol 16(9):1999–2011. CrossRefPubMedGoogle Scholar
  15. Garcia-Carmona JA, Martinez-Laorden E, Milanes MV, Laorden ML (2015) Sympathetic activity induced by naloxone-precipitated morphine withdrawal is blocked in genetically engineered mice lacking functional CRF1 receptor. Toxicol Appl Pharmacol 283(1):42–49. CrossRefPubMedGoogle Scholar
  16. Georgiou P, Zanos P, Ehteramyan M, Hourani S, Kitchen I, Maldonado R, Bailey A (2015) Differential regulation of mGlu5 R and MuOPr by priming- and cue-induced reinstatement of cocaine-seeking behaviour in mice. Addict Biol 20(5):902–912. CrossRefPubMedGoogle Scholar
  17. Georgiou P, Zanos P, Garcia-Carmona JA, Hourani S, Kitchen I, Laorden ML, Bailey A (2016) Methamphetamine abstinence induces changes in u-opioid receptor, oxytocin and CRF systems: association with an anxiogenic phenotype. Neuropharmacology 105:520–532. CrossRefPubMedGoogle Scholar
  18. Gonzales R, Mooney L, Rawson RA (2010) The methamphetamine problem in the United States. Ann Rev Pub Health 31(1):385–398. CrossRefGoogle Scholar
  19. Henry BL, Minassian A, Perry W (2012) Effect of methamphetamine dependence on heart rate variability. Addict Biol 17(3):648–658. CrossRefPubMedGoogle Scholar
  20. Ito H, Yeo KK, Wijetunga M, Seto TB, Tay K, Schatz IJ (2009) A comparison of echocardiographic findings in young adults with cardiomyopathy: with and without a history of methamphetamine abuse. Clin Cardiol 32:18–22CrossRefGoogle Scholar
  21. Kasch S (1987) Serum catecholamines in cocaine intoxicated patients with cardiac symptoms. Ann Emerg Med 16:481Google Scholar
  22. Kaye S, McKetin R, Duflou J, Darke S (2007) Methamphetamine and cardiovascular pathology: a review of the evidence. Addiction 102(8):1204–1211. CrossRefPubMedGoogle Scholar
  23. Kazakova TB, Barabanova SV, Novikova NS, Nosov MA, Rogers VV, Korneva EA (2000) Induction of c-fos and interleukin-2 genes expression in the central nervous system following stressor stimuli. Pathophysiology 7(1):53–61. CrossRefPubMedGoogle Scholar
  24. Koob GF (2010) The role of CRF and CRF-related peptides in the dark side of addiction. Brain Res 1314:3–14. CrossRefPubMedGoogle Scholar
  25. Koob GF, Le Moal M (2008) Neurobiological mechanisms for opponent motivational processes in addiction. Philos Trans R Soc Lond Ser B Biol Sci 363(1507):3113–3123. CrossRefGoogle Scholar
  26. Koob GF, Volkow ND (2016) Neurobiology of addiction: a neurocircuitry analysis. Lancet Psychiatry 3(8):760–773. CrossRefPubMedGoogle Scholar
  27. Krasnova IN, Cadet JL (2009) Methamphetamine toxicity and messengers of death. Brain Res Rev 60(2):379–407. CrossRefPubMedPubMedCentralGoogle Scholar
  28. Latchman DS (2002) Protection of neuronal and cardiac cells by HSP27. Prog Mol Subcell Biol 28:253–265. CrossRefPubMedGoogle Scholar
  29. LaVoie MJ, Hastings TG (1999) Dopamine quinone formation and protein modification associated with the striatal neurotoxicity of methamphetamine: evidence against a role for extracellular dopamine. J Neurosci 19(4):1484–1491PubMedGoogle Scholar
  30. Li T, Chen CK, Hu X, Ball D, Lin SK, Chen W et al (2004) Association analysis of the DRD4 and COMT genes in methamphetamine abuse. Am J Med Genet B Neuropsychiatr Genet 129B(1):120–124. CrossRefPubMedGoogle Scholar
  31. Loogrip ML, Koob GF, Zorrilla EP (2011) Role of corticotropin-releasing factor in drug addiction: potential for pharmacological intervention. CNS Drugs 25(4):271–287. CrossRefGoogle Scholar
  32. Martínez-Laoden E, Hurle MA, Milanés MV, Laorden ML, Almela P (2012) Morphine withdrawal activates hypothalamic–pituitary–adrenal axis and heat shock protein 27 in the left ventricle: the role of extracellular signal regulated kinase. J Pharmacol Exp Ther 342(3):665–675. CrossRefGoogle Scholar
  33. Martinez-Laorden E, Garcia-Carmona JA, Baroja-Mazo A, Romecin P, Atucha NM, Milanes MV, Laorden ML (2014) Corticotropin-releasing factor (CRF) receptor-1 is involved in cardiac noradrenergic activity observed during naloxone-precipitated morphine withdrawal. Br J Pharmacol 171(3):688–700. CrossRefPubMedPubMedCentralGoogle Scholar
  34. Mehlen P, Mehlen A, Godet J, Arrigo AP (1997) Hsp27 as a switch between differentiation and apoptosis in murine embryonic stem cells. J Biol Chem 272(50):31657–31665. CrossRefPubMedGoogle Scholar
  35. Motbey CP, Hunt GE, Bowen MT, Artiss S, McGregor IS (2012) Mephedrone (4-methylmethcathinone, ‘meow’): acute behavioural effects and distribution of Fos expression in adolescent rats. Addict Biol 17(2):409–422. CrossRefPubMedGoogle Scholar
  36. Mymrikov EV, Seit-Nebi AS, Gusev NB (2011) Large potentials of small heat shock proteins. Physiol Rev 91(4):1123–1159. CrossRefPubMedGoogle Scholar
  37. Nash JF, Yamamoto BK (1992) Methamphetamine neurotoxicity and striatal glutamate release: comparison to 3,4-methylenedioxymethamphetamine. Brain Res 581(2):237–243. CrossRefPubMedGoogle Scholar
  38. Nijsen MJ, Croiset G, Stam R, Bruinjnzeel A, Diamant M, de Wied D et al (2000) The role of the CRH type 1 receptor in autonomic responses to corticotropin-releasing hormone in the rat. Neurophsychopharmacology 22(4):388–399. CrossRefGoogle Scholar
  39. Nijsen MJ, Croiset G, Diamant M, De Wied D, Wiegant VM (2001) CRH signalling in the bed nucleus of the stria terminalis is involved in stress-induced cardiac vagal activation in conscious rats. Neuropsychopharmacology 24(1):1–10. CrossRefPubMedGoogle Scholar
  40. Nobis WP, Kash TL, Silberman Y, Winder DG (2011) β-Adrenergic receptors enhance excitatory transmission in the bed nucleus of the stria terminalis through a corticotrophin-releasing factor receptor-dependent and cocaine-regulated mechanism. Biol Psychiatry 69(11):1083–1090. CrossRefPubMedPubMedCentralGoogle Scholar
  41. Nuñez C, Martín F, Földes A, Laorden ML, Kovács KJ, Milanés MV (2010) Induction of FosB/ΔFosB in the brain stress system-related structures during morphine dependence and withdrawal. J Neurochem 114(2):475–487. CrossRefPubMedGoogle Scholar
  42. O’Dell SJ, Weihmuller FB, Marshall JF (1991) Multiple methamphetamine injections induce marked increases in extracellular striatal dopamine which correlate with subsequent neurotoxicity. Brain Res 564(2):256–260. CrossRefPubMedGoogle Scholar
  43. Olive MF, Koenig HN, Nannini MA, Hodge CW (2002) Elevated extracellular CRF levels in the bed nucleus of the stria terminalis during ethanol withdrawal and reduction by subsequent ethanol intake. Pharmacol Biochem Behav 72(1-2):213–220. CrossRefPubMedGoogle Scholar
  44. Oliveira LA, Almeida J, Benini R, Crestani CC (2015) CRF1 and CRF2 receptors in the bed nucleus of the stria terminalis modulate the cardiovascular responses to acute restraint stress in rats. Pharmacol Res 95-96:53–62. CrossRefPubMedGoogle Scholar
  45. Ongur D, An X, Price JL (1998) Prefrontal cortical projections to the hypothalamus in macaque monkeys. J Comp Neurol 401(4):480–505.<480::AID-CNE4>3.0.CO;2-F CrossRefPubMedGoogle Scholar
  46. Panenka WJ, Procyshyn RM, Lecomte T, MacEwan GW, Flynn SW, Honer WG, Barr AM (2013) Methamphetamine use: a comprehensive review of molecular, preclinical and clinical findings. Drug Alcohol Depend 129(3):167–179. CrossRefPubMedGoogle Scholar
  47. Peart JN, Gross GJ (2006) Cardioprotective effects of acute and chronic opioid treatment are mediated via different signaling pathways. Am J Physiol Heart CircPhysiol 291:1746–1753CrossRefGoogle Scholar
  48. Risold PY, Swanson LW (1997) Connections of the rat lateral septal complex. Brain Res Rev 24(2-3):115–195. CrossRefPubMedGoogle Scholar
  49. Robinson AA, Dunn MJ, McCormack A, dos Remedios C, Rose ML (2010) Protective effect of phosphorylated Hsp27 in coronary arteries through actin stabilization. J Mol Cell Cardiol 49(3):370–379. CrossRefPubMedGoogle Scholar
  50. Rogalla T, Ehrnsperger M, Preville X, Kotlyarov A, Lutsch G, Ducasse C, Paul C, Wieske M, Arrigo AP, Buchner J, Gaestel M (1999) Regulation of Hsp27 oligomerization, chaperone function, and protective activity against oxidative stress/tumor necrosis factor alpha by phosphorylation. J BiolChem 274:18947–18956Google Scholar
  51. Sawchenko PE, Swanson LW (1982) Immunohistochemical identification of neurons in the paraventricular nucleus of the hypothalamus that project to the medulla or to the spinal cord in the rat. J Comp Neurol 205(3):260–272. CrossRefPubMedGoogle Scholar
  52. Sharko AC, Kaigler KF, Fadel JR, Wilson MA (2016) Ethanol-induced anxiolysis and neuronal activation in the amygdala and bed nucleus of the stria terminalis. Alcohol 50:19–25. CrossRefPubMedGoogle Scholar
  53. Stamatakis AM, Sparta DR, Jennings JH, McElligott ZA, Decot H, Stuber GD (2014) Amygdala and bed nucleus of the stria terminalis circuitry: implications for addiction-related behaviors. Neuropharmacology 76:320–328. CrossRefPubMedGoogle Scholar
  54. Stephans SE, Yamamoto BK (1994) Methamphetamine-induced neurotoxicity: roles for glutamate and dopamine efflux. Synapse 17(3):203–209. CrossRefPubMedGoogle Scholar
  55. Sulzer D, Sonders MS, Poulsen NW, Galli A (2005) Mechanisms of neurotransmitter release by amphetamines: a review. Prog Neurobiol 75(6):406–433. CrossRefPubMedGoogle Scholar
  56. Toth ME, Gonda S, Vigh L, Santha M (2010) Neuroprotective effect of small heat shock protein, Hsp27, after acute and chronic alcohol administration. Cell Stress Chaperones 15(6):807–817. CrossRefPubMedPubMedCentralGoogle Scholar
  57. Tunbridge EM, Huber A, Farrell SM, Stumpenhorst K, Harrison PJ, Walton ME (2013) The role of cateho-o-methyltransferase in reward processing and addiction. CNS Neurol Disord Drug Targets 11:306–323CrossRefGoogle Scholar
  58. Wada K (2011) The history and current state of drug abuse in Japan. Ann N Y Acad Sci 1216(1):62–72. CrossRefPubMedGoogle Scholar
  59. Wagner GC, Ricaurte GA, Seiden LS, Schuster CR, Miller RJ, Westley J (1980) Long-lasting depletions of striatal dopamine and loss of dopamine uptake sites following repeated administration of methamphetamine. Brain Res 181(1):151–160. CrossRefPubMedGoogle Scholar
  60. Wiechelman KJ, Braun RD, Fitpatrick JD (1988) Investigation of the bicinchoninic acid protein assay: identification of the groups responsible for color formation. Anal Biochem 175(1):231–237. CrossRefPubMedGoogle Scholar
  61. Yamamoto BK, Zhu W (1998) The effects of methamphetamine on the production of free radicals and oxidative stress. J Pharmacol Exp Ther 287(1):107–114PubMedGoogle Scholar
  62. Yang LZ, Tovote P, Rayner M, Kockskämper J, Pieske B, Spiess J (2010) Corticotropin-releasing factor receptors and urocortins, links between the brain and the heart. Eur J Pharmacol 632(1-3):1–6. CrossRefPubMedGoogle Scholar
  63. Yu Y, Wei SG, Zhang ZH, Weiss RM, Felder RB (2016) ERK1/2 MAPK signaling in hypothalamic paraventricular nucleus contributes to sympathetic excitation in rats with heart failure after myocardial infarction. Am J Physiol Heart Circ Physiol 310(6):H732–H739. CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

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

Authors and Affiliations

  1. 1.Department of Pharmacology, Faculty of MedicineUniversity of MurciaMurciaSpain
  2. 2.Unit of Acute PsychiatryReina Sofía University HospitalMurciaSpain
  3. 3.Department of Biochemistry, Faculty of Health and Medical SciencesUniversity of SurreyGuildfordUK
  4. 4.Department of PsychiatryUniversity of Maryland School of MedicineBaltimoreUSA
  5. 5.Institute of Medical and Biomedical EducationSt. George’s University of LondonLondonUK

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