, Volume 232, Issue 19, pp 3539–3549 | Cite as

Antidepressant-like effects of guanfacine and sex-specific differences in effects on c-fos immunoreactivity and paired-pulse ratio in male and female mice

  • Yann S. Mineur
  • Matthew P. Bentham
  • Wen-Liang Zhou
  • Margreet E. Plantenga
  • Sherry A. McKee
  • Marina R. PicciottoEmail author
Original Investigation



The a2A-noradrenergic agonist guanfacine can decreases stress-induced smoking in female, but not male, human smokers. It is not known whether these effects are due to effects on mood regulation and/or result from nicotinic-cholinergic interactions.


The objective of the study was to determine whether there are sex differences in the effect of guanfacine in tests of anxiolytic and antidepressant efficacy in mice at baseline and in a hypercholinergic model of depression induced by the acetylcholinesterase inhibitor physostigmine.


The effects of guanfacine were measured in the light/dark box, tail suspension, and the forced swim test in female and male C57BL/6J mice. In parallel, electrophysiological properties were evaluated in the prefrontal cortex, a critical brain region involved in stress responses. c-fos immunoreactivity was measured in other brain regions known to regulate mood.


Despite a baseline sex difference in behavior in the forced swim test (female mice were more immobile), guanfacine had similar, dose-dependent, antidepressant-like effects in mice of both sexes (optimal dose, 0.15 mg/kg). An antidepressant-like effect of guanfacine was also observed following pre-treatment with physostigmine. A sex difference in the paired-pulse ratio in the prefrontal cortex (PFC) (male, 1.4; female, 2.1) was observed at baseline that was normalized by guanfacine. Other brain areas involved in cholinergic control of depression-like behaviors, including the basolateral amygdala and lateral septum, showed sex-specific changes in c-fos expression.


Guanfacine has a robust antidepressant-like effect and can reverse a depression-like state induced by increased acetylcholine (ACh) signaling. These data suggest that different brain areas are recruited in female and male mice, despite similar behavioral responses to guanfacine.


Norepinephrine Sex difference Depression Amygdala Prefrontal cortex Mouse models 



This work was supported by P50 DA033945 (ORWH, NIDA, FDA), MH077681, and MH105824.

Conflict of interest

The authors declare that they have no competing interests.


  1. Alarcon G, Cservenka A, Rudolph MD, Fair DA, Nagel BJ (2015) Developmental sex differences in resting state functional connectivity of amygdala sub-regions. Neuroimage 115:235–44CrossRefPubMedGoogle Scholar
  2. Aston-Jones G, Kalivas PW (2008) Brain norepinephrine rediscovered in addiction research. Biol Psychiatry 63:1005–6PubMedCentralCrossRefPubMedGoogle Scholar
  3. Beiranvand F, Zlabinger C, Orr-Urtreger A, Ristl R, Huck S, Scholze P (2014) Nicotinic acetylcholine receptors control acetylcholine and noradrenaline release in the rodent habenulo- interpeduncular complex. Br J Pharmacol 171:5209–24PubMedCentralCrossRefPubMedGoogle Scholar
  4. Bourin M, Colombel MC, Malinge M, Bradwejn J (1991) Clonidine as a sensitizing agent in the forced swimming test for revealing antidepressant activity. J Psychiatry Neurosci 16:199–203PubMedCentralPubMedGoogle Scholar
  5. Carcoba LM, Orfila JE, Natividad LA, Torres OV, Pipkin JA, Ferree PL, Castaneda E, Moss DE, O'Dell LE (2014) Cholinergic transmission during nicotine withdrawal is influenced by age and pre-exposure to nicotine: implications for teenage smoking. Dev Neurosci 36:347–55PubMedCentralCrossRefPubMedGoogle Scholar
  6. Carrier GO, Bishop VS (1972) The interaction of acetylcholine and norepinephrine on heart rate. J Pharmacol Exp Ther 180:31–7PubMedGoogle Scholar
  7. Dell'Osso B, Palazzo MC, Oldani L, Altamura AC (2011) The noradrenergic action in antidepressant treatments: pharmacological and clinical aspects. CNS Neurosci Ther 17:723–32CrossRefPubMedGoogle Scholar
  8. Devore S, Linster C (2012) Noradrenergic and cholinergic modulation of olfactory bulb sensory processing. Front Behav Neurosci 6:52PubMedCentralPubMedGoogle Scholar
  9. Dilsaver SC (1986) Cholinergic mechanisms in depression. Brain Res 396:285–316CrossRefPubMedGoogle Scholar
  10. Esterlis I, Hannestad JO, Bois F, Sewell RA, Tyndale RF, Seibyl JP, Picciotto MR, Laruelle M, Carson RE, Cosgrove KP (2013a) Imaging changes in synaptic acetylcholine availability in living human subjects. J Nucl Med 54:78–82PubMedCentralCrossRefPubMedGoogle Scholar
  11. Esterlis I, Ranganathan M, Bois F, Pittman B, Picciotto MR, Shearer L, Anticevic A, Carlson J, Niciu MJ, Cosgrove KP, D'Souza DC (2013b) In vivo evidence for beta 2 nicotinic acetylcholine receptor subunit upregulation in smokers as compared with nonsmokers with schizophrenia. Biological PsychiatryGoogle Scholar
  12. Fenster CP, Whitworth TL, Sheffield EB, Quick MW, Lester RA (1999) Upregulation of surface alpha4beta2 nicotinic receptors is initiated by receptor desensitization after chronic exposure to nicotine. J Neurosci 19:4804–14PubMedGoogle Scholar
  13. Fox H, Sinha R (2014) The role of guanfacine as a therapeutic agent to address stress-related pathophysiology in cocaine-dependent individuals. Adv Pharmacol 69:217–65PubMedCentralCrossRefPubMedGoogle Scholar
  14. Fox HC, Morgan PT, Sinha R (2014) Sex differences in guanfacine effects on drug craving and stress arousal in cocaine-dependent individuals. Neuropsychopharmacology 39:1527–37PubMedCentralCrossRefPubMedGoogle Scholar
  15. Fox H, Sofuoglu M, Sinha R (2015a) Guanfacine enhances inhibitory control and attentional shifting in early abstinent cocaine-dependent individuals. J Psychopharmacol 29:312–23PubMedCentralCrossRefPubMedGoogle Scholar
  16. Fox ME, Studebaker RI, Swofford NJ, Wightman RM (2015b) Stress and drug dependence differentially modulate norepinephrine signaling in animals with varied HPA axis function. Neuropsychopharmacology 40(7):1752–61CrossRefPubMedGoogle Scholar
  17. Glavin GB (1985) Stress and brain noradrenaline: a review. Neurosci Biobehav Rev 9:233–43CrossRefPubMedGoogle Scholar
  18. Gold PW (2015) The organization of the stress system and its dysregulation in depressive illness. Mol Psychiatry 20:32–47CrossRefPubMedGoogle Scholar
  19. Hains AB, Vu MA, Maciejewski PK, van Dyck CH, Gottron M, Arnsten AF (2009) Inhibition of protein kinase C signaling protects prefrontal cortex dendritic spines and cognition from the effects of chronic stress. Proc Natl Acad Sci U S A 106:17957–62PubMedCentralCrossRefPubMedGoogle Scholar
  20. Hains AB, Yabe Y, Arnsten AF (2015) Chronic stimulation of alpha-2a-adrenoceptors with guanfacine protects rodent prefrontal cortex dendritic spines and cognition from the effects of chronic stress. Neurobiol Stress 2:1–9CrossRefPubMedGoogle Scholar
  21. Hannestad JO, Cosgrove KP, DellaGioia NF, Perkins E, Bois F, Bhagwagar Z, Seibyl JP, McClure-Begley TD, Picciotto MR, Esterlis I (2013) Changes in the cholinergic system between bipolar depression and euthymia as measured with [123I]5IA single photon emission computed tomography. Biol Psychiatry 74:768–76PubMedCentralCrossRefPubMedGoogle Scholar
  22. Janowsky DS, Overstreet DH (1990a) Cholinergic dysfunction in depression. Pharmacol Toxicol 3:100–11CrossRefGoogle Scholar
  23. Janowsky DS, Overstreet DH (1990b) Cholinergic dysfunction in depression. Pharmacol Toxicol 66(Suppl 3):100–11CrossRefPubMedGoogle Scholar
  24. Janowsky DS, El-Yousef MK, Davis JM, Sekerke HJ (1972) A cholinergic-adrenergic hypothesis of mania and depression. Lancet 2:632–5CrossRefPubMedGoogle Scholar
  25. Kaufer D, Friedman A, Seidman S, Soreq H (1998) Acute stress facilitates long-lasting changes in cholinergic gene expression. Nature 393:373–7CrossRefPubMedGoogle Scholar
  26. Martinowich K, Schloesser RJ, Lu Y, Jimenez DV, Paredes D, Greene JS, Greig NH, Manji HK, Lu B (2012) Roles of p75(NTR), long-term depression, and cholinergic transmission in anxiety and acute stress coping. Biol Psychiatry 71:75–83PubMedCentralCrossRefPubMedGoogle Scholar
  27. McElligott ZA, Fox ME, Walsh PL, Urban DJ, Ferrel MS, Roth BL, Wightman RM (2013) Noradrenergic synaptic function in the bed nucleus of the stria terminalis varies in animal models of anxiety and addiction. Neuropsychopharmacology 38:1665–73PubMedCentralCrossRefPubMedGoogle Scholar
  28. McKee SA, Potenza MN, Kober H, Sofuoglu M, Arnsten AF, Picciotto MR, Weinberger AH, Ashare R, Sinha R (2015) A translational investigation targeting stress-reactivity and prefrontal cognitive control with guanfacine for smoking cessation. J Psychopharmacol 29:300–11PubMedCentralCrossRefPubMedGoogle Scholar
  29. Milusheva E, Baranyi M, Zelles T, Mike A, Vizi ES (1994) Release of acetylcholine and noradrenaline from the cholinergic and adrenergic afferents in rat hippocampal CA1, CA3 and dentate gyrus regions. Eur J Neurosci 6:187–92CrossRefPubMedGoogle Scholar
  30. Mineur YS, Somenzi O, Picciotto MR (2007) Cytisine, a partial agonist of high-affinity nicotinic acetylcholine receptors, has antidepressant-like properties in male C57BL/6J mice. Neuropharmacology 52:1256–62PubMedCentralCrossRefPubMedGoogle Scholar
  31. Mineur YS, Obayemi A, Wigestrand MB, Fote GM, Calarco CA, Li AM, Picciotto MR (2013) Cholinergic signaling in the hippocampus regulates social stress resilience and anxiety- and depression-like behavior. Proc Natl Acad Sci U S A 110:3573–8PubMedCentralCrossRefPubMedGoogle Scholar
  32. Rabenstein RL, Caldarone BJ, Picciotto MR (2006) The nicotinic antagonist mecamylamine has antidepressant-like effects in wild-type but not beta2- or alpha7-nicotinic acetylcholine receptor subunit knockout mice. Psychopharmacology 189:395–401CrossRefPubMedGoogle Scholar
  33. Risch SC, Cohen RM, Janowsky DS, Kalin NH, Murphy DL (1980) Mood and behavioral effects of physostigmine on humans are accompanied by elevations in plasma beta-endorphin and cortisol. Science 209:1545–6CrossRefPubMedGoogle Scholar
  34. Risch SC, Cohen RM, Janowsky DS, Kalin NH, Sitaram N, Gillin JC, Murphy DL (1981) Physostigmine induction of depressive symptomatology in normal human subjects. Psychiatry Res 4:89–94CrossRefPubMedGoogle Scholar
  35. Saricicek A, Esterlis I, Maloney KH, Mineur YS, Ruf BM, Muralidharan A, Chen JI, Cosgrove KP, Kerestes R, Ghose S, Tamminga CA, Pittman B, Bois F, Tamagnan G, Seibyl J, Picciotto MR, Staley JK, Bhagwagar Z (2012) Persistent beta2*-nicotinic acetylcholinergic receptor dysfunction in major depressive disorder. Am J Psychiatry 169:851–9PubMedCentralCrossRefPubMedGoogle Scholar
  36. Schulz KP, Clerkin SM, Fan J, Halperin JM, Newcorn JH (2013) Guanfacine modulates the influence of emotional cues on prefrontal cortex activation for cognitive control. Psychopharmacology 226:261–71PubMedCentralCrossRefPubMedGoogle Scholar
  37. Sepede G, Corbo M, Fiori F, Martinotti G (2012) Reboxetine in clinical practice: a review. La Clinica Ter 163:e255–62Google Scholar
  38. Shaltiel G, Hanan M, Wolf Y, Barbash S, Kovalev E, Shoham S, Soreq H (2013) Hippocampal microRNA-132 mediates stress-inducible cognitive deficits through its acetylcholinesterase target. Brain Struct Funct 218:59–72PubMedCentralCrossRefPubMedGoogle Scholar
  39. Sillence MN, Tudor GD, Matthews ML, Lindsay DB (1992) Effects of the alpha 2-adrenoceptor agonist guanfacine on growth and thermogenesis in mice. J Anim Sci 70:3429–34PubMedGoogle Scholar
  40. Sofuoglu M, Rosenheck R, Petrakis I (2014) Pharmacological treatment of comorbid PTSD and substance use disorder: recent progress. Addict Behav 39:428–33CrossRefPubMedGoogle Scholar
  41. Sorkin EM, Heel RC (1986) Guanfacine. A review of its pharmacodynamic and pharmacokinetic properties, and therapeutic efficacy in the treatment of hypertension. Drugs 31:301–36CrossRefPubMedGoogle Scholar
  42. Verplaetse TL, Weinberger AH, Smith PH, Cosgrove KP, Mineur YS, Picciotto MR, Mazure CM, McKee SA (2015) Targeting the noradrenergic system for gender-sensitive medication development for tobacco dependence. Nicotine Tob Res 17:486–495CrossRefPubMedGoogle Scholar
  43. Weinberger AH, Smith PH, Kaufman M, McKee SA (2014) Consideration of sex in clinical trials of transdermal nicotine patch: a systematic review. Exp Clin Psychopharmacol 22:373–83CrossRefPubMedGoogle Scholar
  44. Wilkinson DS, Turner JR, Blendy JA, Gould TJ (2013) Genetic background influences the effects of withdrawal from chronic nicotine on learning and high-affinity nicotinic acetylcholine receptor binding in the dorsal and ventral hippocampus. Psychopharmacology 225:201–8PubMedCentralCrossRefPubMedGoogle Scholar
  45. Zaborszky L, Rosin DL, Kiss J (2004) Alpha-adrenergic receptor alpha2A is colocalized in basal forebrain cholinergic neurons: a light and electron microscopic double immunolabeling study. J Neurocytol 33:265–76CrossRefPubMedGoogle Scholar
  46. Zhou WL, Antic SD (2012) Rapid dopaminergic and GABAergic modulation of calcium and voltage transients in dendrites of prefrontal cortex pyramidal neurons. J Physiol 590:3891–911PubMedCentralCrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Yann S. Mineur
    • 1
  • Matthew P. Bentham
    • 1
  • Wen-Liang Zhou
    • 1
  • Margreet E. Plantenga
    • 1
  • Sherry A. McKee
    • 1
  • Marina R. Picciotto
    • 1
    Email author
  1. 1.Department of PsychiatryYale University School of MedicineNew HavenUSA

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