Archival Report| Volume 70, ISSUE 10, P920-927, November 15, 2011

Estradiol Modulates Medial Prefrontal Cortex and Amygdala Activity During Fear Extinction in Women and Female Rats

  • Mohamed A. Zeidan
    Department of Psychiatry, Massachusetts General Hospital, Boston; Harvard Medical School, Charlestown, Massachusetts
    Search for articles by this author
  • Sarah A. Igoe
    Department of Psychiatry, Massachusetts General Hospital, Boston; Harvard Medical School, Charlestown, Massachusetts
    Search for articles by this author
  • Clas Linnman
    Department of Psychiatry, Massachusetts General Hospital, Boston; Harvard Medical School, Charlestown, Massachusetts
    Search for articles by this author
  • Antonia Vitalo
    Department of Psychiatry, Massachusetts General Hospital, Boston; Harvard Medical School, Charlestown, Massachusetts
    Search for articles by this author
  • John B. Levine
    Department of Psychiatry, Massachusetts General Hospital, Boston; Harvard Medical School, Charlestown, Massachusetts

    The Benson Henry Institute for Mind Body Medicine, Massachusetts General Hospital, Boston, Massachusetts

    Shriners Burns Hospital for Children, Boston, Massachusetts
    Search for articles by this author
  • Anne Klibanski
    Neuroendocrine Unit, Massachusetts General Hospital, Boston, Massachusetts

    Harvard Medical School, Boston, Massachusetts
    Search for articles by this author
  • Jill M. Goldstein
    Department of Psychiatry, Massachusetts General Hospital, Boston; Harvard Medical School, Charlestown, Massachusetts

    Departments of Psychiatry and Medicine, Division of Women's Health, Brigham and Women's Hospital and Harvard Medical School, Boston, Massachusetts
    Search for articles by this author
  • Mohammed R. Milad
    Address correspondence to Mohammed R. Milad, Ph.D., Department of Psychiatry, Harvard Medical School and Massachusetts General Hospital, 149 13th Street, CNY 2614, Charlestown, MA 02129
    Department of Psychiatry, Massachusetts General Hospital, Boston; Harvard Medical School, Charlestown, Massachusetts
    Search for articles by this author


      Men and women differ in their ability to extinguish fear. Fear extinction requires the activation of brain regions, including the ventromedial prefrontal cortex (vmPFC) and amygdala. Could estradiol modulate the activity of these brain regions during fear extinction?


      All rat experiments were conducted in naturally cycling females. Rats underwent fear conditioning on Day 1. On Day 2, they underwent extinction training during the metestrus phase of the cycle (low estrogen and progesterone). Extinction recall was assessed on Day 3. Systemic injections of estrogen receptor-beta and -alpha agonists and of estradiol were administered at different time points to assess their influence on extinction consolidation and c-Fos expression in the vmPFC and amygdala. In parallel, healthy naturally cycling women underwent an analogous fear conditioning extinction training in a 3T functional magnetic resonance scanner. Measurement of their estradiol levels and skin conductance responses were obtained throughout the experiment.


      In female rats, administration of the estrogen-receptor beta (but not alpha) agonist facilitated extinction recall. Immediate (but not delayed) postextinction training administration of estradiol facilitated extinction memory consolidation and increased c-Fos expression in the vmPFC while reducing it in the amygdala. In parallel, natural variance in estradiol in premenopausal cycling women modulated vmPFC and amygdala reactivity and facilitated extinction recall.


      We provide translational evidence that demonstrates the influence of endogenous and exogenous estradiol on the fear extinction network. Our data suggest that women's endogenous hormonal status should be considered in future neurobiological research related to anxiety and mood disorders.

      Key Words

      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'


      Subscribe to Biological Psychiatry
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect


        • Sotres-Bayon F.
        • Quirk G.J.
        Prefrontal control of fear: More than just extinction.
        Curr Opin Neurobiol. 2010; 20: 231-235
        • Herry C.
        • Ferraguti F.
        • Singewald N.
        • Letzkus J.J.
        • Ehrlich I.
        • Luthi A.
        Neuronal circuits of fear extinction.
        Eur J Neurosci. 2010; 31: 599-612
        • Schiller D.
        • Delgado M.R.
        Overlapping neural systems mediating extinction, reversal and regulation of fear.
        Trends Cogn Sci. 2010; 14: 268-276
        • Quirk G.J.
        • Mueller D.
        Neural mechanisms of extinction learning and retrieval.
        Neuropsychopharmacology. 2008; 33: 56-72
        • Milad M.R.
        • Rauch S.L.
        • Pitman R.K.
        • Quirk G.J.
        Fear extinction in rats: Implications for human brain imaging and anxiety disorders.
        Biol Psychol. 2006; 73: 61-71
        • Milad M.R.
        • Vidal-Gonzalez I.
        • Quirk G.J.
        Electrical stimulation of medial prefrontal cortex reduces conditioned fear in a temporally specific manner.
        Behav Neurosci. 2004; 118: 389-394
        • Herry C.
        • Ciocchi S.
        • Senn V.
        • Demmou L.
        • Muller C.
        • Luthi A.
        Switching on and off fear by distinct neuronal circuits.
        Nature. 2008; 454: 600-606
        • Likhtik E.
        • Popa D.
        • Apergis-Schoute J.
        • Fidacaro G.A.
        • Pare D.
        Amygdala intercalated neurons are required for expression of fear extinction.
        Nature. 2008; 454: 642-645
        • Milad M.R.
        • Quirk G.J.
        Neurons in medial prefrontal cortex signal memory for fear extinction.
        Nature. 2002; 420: 70-74
        • Milad M.R.
        • Wright C.I.
        • Orr S.P.
        • Pitman R.K.
        • Quirk G.J.
        • Rauch S.L.
        Recall of fear extinction in humans activates the ventromedial prefrontal cortex and hippocampus in concert.
        Biol Psychiatry. 2007; 62: 446-454
        • Milad M.R.
        • Quinn B.T.
        • Pitman R.K.
        • Orr S.P.
        • Fischl B.
        • Rauch S.L.
        Thickness of ventromedial prefrontal cortex in humans is correlated with extinction memory.
        Proc Natl Acad Sci U S A. 2005; 102: 10706-10711
        • Phelps E.A.
        • Delgado M.R.
        • Nearing K.I.
        • Ledoux J.E.
        Extinction learning in humans: Role of the amygdala and vmPFC.
        Neuron. 2004; 43: 897-905
        • Sehlmeyer C.
        • Schoning S.
        • Zwitserlood P.
        • Pfleiderer B.
        • Kircher T.
        • Arolt V.
        • Konrad C.
        Human fear conditioning and extinction in neuroimaging: A systematic review.
        PLoS ONE. 2009; 4: e5865
        • LaBar K.S.
        • Gatenby J.C.
        • Gore J.C.
        • Ledoux J.E.
        • Phelps E.A.
        Human amygdala activation during conditioned fear acquisition and extinction: A mixed-trial fMRI study.
        Neuron. 1998; 20: 937-945
        • Knight D.C.
        • Nguyen H.T.
        • Bandettini P.A.
        The role of the human amygdala in the production of conditioned fear responses.
        Neuroimage. 2005; 26: 1193-1200
        • Kinrys G.
        • Wygant L.E.
        Anxiety disorders in women: Does gender matter to treatment?.
        Rev Bras Psiquiatr. 2005; 27: S43-S50
        • Milad M.R.
        • Pitman R.K.
        • Ellis C.B.
        • Gold A.L.
        • Shin L.M.
        • Lasko N.B.
        • et al.
        Neurobiological basis of failure to recall extinction memory in posttraumatic stress disorder.
        Biol Psychiatry. 2009; 66: 1075-1082
        • Frye C.A.
        • Edinger K.
        • Sumida K.
        Androgen administration to aged male mice increases anti-anxiety behavior and enhances cognitive performance.
        Neuropsychopharmacology. 2008; 33: 1049-1061
        • Smith C.C.
        • Vedder L.C.
        • McMahon L.L.
        Estradiol and the relationship between dendritic spines, NR2B containing NMDA receptors, and the magnitude of long-term potentiation at hippocampal CA3-CA1 synapses.
        Psychoneuroendocrinology. 2009; 34: S130-S142
        • Milad M.R.
        • Goldstein J.M.
        • Orr S.P.
        • Wedig M.M.
        • Klibanski A.
        • Pitman R.K.
        • Rauch S.L.
        Fear conditioning and extinction: Influence of sex and menstrual cycle in healthy humans.
        Behav Neurosci. 2006; 120: 1196-1203
        • Leuner B.
        • Mendolia-Loffredo S.
        • Shors T.J.
        High levels of estrogen enhance associative memory formation in ovariectomized females.
        Psychoneuroendocrinology. 2004; 29: 883-890
        • Jackson L.R.
        • Robinson T.E.
        • Becker J.B.
        Sex differences and hormonal influences on acquisition of cocaine self-administration in rats.
        Neuropsychopharmacology. 2006; 31: 129-138
        • Goldstein J.M.
        • Seidman L.J.
        • Horton N.J.
        • Makris N.
        • Kennedy D.N.
        • Caviness Jr, V.S.
        • et al.
        Normal sexual dimorphism of the adult human brain assessed by in vivo magnetic resonance imaging.
        Cereb Cortex. 2001; 11: 490-497
        • Ostlund H.
        • Keller E.
        • Hurd Y.L.
        Estrogen receptor gene expression in relation to neuropsychiatric disorders.
        Ann N Y Acad Sci. 2003; 1007: 54-63
        • Andreano J.M.
        • Cahill L.
        Sex influences on the neurobiology of learning and memory.
        Learn Mem. 2009; 16: 248-266
        • Goldstein J.M.
        • Jerram M.
        • Poldrack R.
        • Ahern T.
        • Kennedy D.N.
        • Seidman L.J.
        • Makris N.
        Hormonal cycle modulates arousal circuitry in women using functional magnetic resonance imaging.
        J Neurosci. 2005; 25: 9309-9316
        • Dalla C.
        • Shors T.J.
        Sex differences in learning processes of classical and operant conditioning.
        Physiol Behav. 2009; 97: 229-238
        • Milad M.R.
        • Igoe S.A.
        • Lebron-Milad K.
        • Novales J.E.
        Estrous cycle phase and gonadal hormones influence conditioned fear extinction.
        Neuroscience. 2009; 164: 887-895
        • Milad M.R.
        • Zeidan M.A.
        • Contero A.
        • Pitman R.K.
        • Klibanski A.
        • Rauch S.L.
        • Goldstein J.M.
        The influence of gonadal hormones on conditioned fear extinction in healthy humans.
        Neuroscience. 2010; 168: 652-658
        • Becker J.B.
        • Arnold A.P.
        • Berkley K.J.
        • Blaustein J.D.
        • Eckel L.A.
        • Hampson E.
        • et al.
        Strategies and methods for research on sex differences in brain and behavior.
        Endocrinology. 2005; 146: 1650-1673
        • Paxinos G.
        • Watson C.
        The Rat Brain in Stereotaxic Coordinates.
        in: 6th edition. Academic Press, San Diego, CA2007
        • Levine J.B.
        • Leeder A.D.
        • Parekkadan B.
        • Berdichevsky Y.
        • Rauch S.L.
        • Smoller J.W.
        • et al.
        Isolation rearing impairs wound healing and is associated with increased locomotion and decreased immediate early gene expression in the medial prefrontal cortex of juvenile rats.
        Neuroscience. 2008; 151: 589-603
        • First M.B.
        • Spitzer R.L.
        • Gibbon M.
        • Williams J.B.W.
        Structured Clinical Interview for DSM-IV-TR Axis I Disorders.
        Biometrics Research, New York Psychiatric Institute, New York2002
        • Milad M.R.
        • Quirk G.J.
        • Pitman R.K.
        • Orr S.P.
        • Fischl B.
        • Rauch S.L.
        A role for the human dorsal anterior cingulate cortex in fear expression.
        Biol Psychiatry. 2007; 62: 1191-1194
        • Talairach J.
        • Tournoux P.
        Co-planar Stereotaxic Atlas of the Human Brain.
        Thieme, New York1988
        • Chang Y.J.
        • Yang C.H.
        • Liang Y.C.
        • Yeh C.M.
        • Huang C.C.
        • Hsu K.S.
        Estrogen modulates sexually dimorphic contextual fear extinction in rats through estrogen receptor beta.
        Hippocampus. 2009; 19: 1142-1150
        • Weiser M.J.
        • Foradori C.D.
        • Handa R.J.
        Estrogen receptor beta activation prevents glucocorticoid receptor-dependent effects of the central nucleus of the amygdala on behavior and neuroendocrine function.
        Brain Res. 2010; 1336: 78-88
        • Greer P.L.
        • Greenberg M.E.
        From synapse to nucleus: Calcium-dependent gene transcription in the control of synapse development and function.
        Neuron. 2008; 59: 846-860
        • Osborne D.M.
        • Edinger K.
        • Frye C.A.
        Chronic administration of androgens with actions at estrogen receptor beta have anti-anxiety and cognitive-enhancing effects in male rats.
        Age (Dordr). 2009; 31: 191-198
        • Tomihara K.
        • Soga T.
        • Nomura M.
        • Korach K.S.
        • Gustafsson J.A.
        • Pfaff D.W.
        • Ogawa S.
        Effect of ER-beta gene disruption on estrogenic regulation of anxiety in female mice.
        Physiol Behav. 2009; 96: 300-306
        • Smith C.C.
        • McMahon L.L.
        Estrogen-induced increase in the magnitude of long-term potentiation occurs only when the ratio of NMDA transmission to AMPA transmission is increased.
        J Neurosci. 2005; 25: 7780-7791
        • Santini E.
        • Muller R.U.
        • Quirk G.J.
        Consolidation of extinction learning involves transfer from NMDA-independent to NMDA-dependent memory.
        J Neurosci. 2001; 21: 9009-9017
        • Santini E.
        • Quirk G.J.
        • Porter J.T.
        Fear conditioning and extinction differentially modify the intrinsic excitability of infralimbic neurons.
        J Neurosci. 2008; 28: 4028-4036
        • Myers K.M.
        • Davis M.
        Mechanisms of fear extinction.
        Mol Psychiatry. 2007; 12: 120-150
        • Llaneza D.C.
        • Frye C.A.
        Progestogens and estrogen influence impulsive burying and avoidant freezing behavior of naturally cycling and ovariectomized rats.
        Pharmacol Biochem Behav. 2009; 93: 337-342
        • Taneepanichskul S.
        • Patrachai S.
        Effects of long-term treatment with depot medroxy progesterone acetate for contraception on estrogenic activity.
        J Med Assoc Thai. 1998; 81: 944-946
        • Altshuler L.L.
        • Hendrick V.
        • Cohen L.S.
        Course of mood and anxiety disorders during pregnancy and the postpartum period.
        J Clin Psychiatry. 1998; 59: 29-33
        • Schnatz P.F.
        • Whitehurst S.K.
        • O'Sullivan D.M.
        Sexual dysfunction, depression, and anxiety among patients of an inner-city menopause clinic.
        J Womens Health (Larchmt). 2010; 19: 1843-1849

      Linked Article

      • Complex Roles of Estrogen in Emotion: Sex Matters
        Biological PsychiatryVol. 70Issue 10
        • Preview
          Women are more than twice as likely to suffer from fear and anxiety disorders, including generalized anxiety disorder, panic attacks, and posttraumatic stress disorder. One reason for this difference may be that fluctuating ovarian hormone levels in women during their reproductive life span alters emotional processing. For example, the incidence of anxiety symptoms is higher when ovarian hormone levels are low, including during premenstrual, postpartum, and perimenopausal periods. These women can benefit from estrogen treatment, suggesting an anxiolytic effect.
        • Full-Text
        • PDF