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Declarative Memory after Stress in Humans: DifferentialInvolvement of the
-Adrenergic andCorticosteroid Systems
Franc¸oise S. Maheu, Ridha Joober, and Sonia J. Lupien
 Laboratory of Human Stress Research (F.S.M., R.J., S.J.L.), Douglas Hospital Research Centre, Montre´al, Que´bec, Canada H4H 1R3; Department of Psychology (F.S.M.), University of Montre´al, Montre´al, Que´bec, Canada H3C 3J7; and Departmentof Psychiatry (R.J., S.J.L.), McGill University, Montre´al, Que´bec, Canada H3A 1A1
To determine the role played by the
-adrenergic and corti-costeroid systems in the modulatory effects of stress on de-clarative memory function, 42 young men were administeredaplacebo,propranolol(
-adrenergicblocker),ormetyrapone(corticosteroid synthesis inhibitor) before beingsubmitted toa psychological stress protocol. Immediately after stress, sub- jects viewed a neutral story, unrelated to the stressor. Short-(5minpostlearning)andlong-term(1wkpostlearning)recallof the story was assessed. Placebo and propranolol groupsshowed significant stress-related increases in corticosteroidlevels,whereasmetyraponepreventedcorticosteroidreactiv-ity to the stressor. Stress triggered significant elevations incardiac activity (heart rate, systolic and diastolic blood pres-sure levels) in all three groups, with the metyrapone groupshowing the strongest elevation in heart rate levels in re-sponse to stress. Compared with placebo, propranolol had noeffect on short- and long-term recall of the story learned afterstress, whereas metyrapone impaired short-term recall of thestory, with no further effects on long-term declarative mem-ory. These results suggest that, contrary to the
-adrenergicsystem, the corticosteroid system is implicated in declarativememory function after stress in humans. (
 J Clin Endocrinol Metab
90: 1697–1704, 2005)
T
HE PHYSIOLOGICAL RESPONSE of an organism toacutestressenablesittocopeefficientlywiththethreat.Duringthisprocess,animals(1–3)andhumans(4,5)developvivid memories of the stressful and emotionally arousingsituation. Interestingly, stressful and emotionally arousingexperiences have also been shown to modulate subsequentdeclarative memory for material unrelated to the source ofthe stressor (
e.g.
Refs. 6–11; for a review, see Refs. 12–14).Declarative memory refers to the conscious or voluntaryrecollection of previously learned information (15). The ef-fectsofstressonsubsequentdeclarativememoryformaterialunrelated to the stressor can have important implications ineverydaylife,giventhepresenceandreoccurrenceofvarioussources of stress in the modern environment. The
-adren-ergic and corticosteroid systems have been studied in rela-tion to the memory-modulating effects of stressful and emo-tionally arousing experiences. The rationale for studying theimpactofthesehormonalsystemsisthatbothsystems,whenactivated during stressful and emotionally arousing events,modulate activity in the frontal lobes, hippocampus, andamygdala, three brain structures involved in declarativememory processes (1, 2, 12–14, 16, 17).In humans, the activation of the corticosteroid system has been shown to modulate poststress memory performance.Studies that have assessed the effects of stress on subsequentmemory processing of material unrelated to the stressor re-ported that stress sometimes enhances (6), has no effect (18,19), or sometimes impairs memory performance (9–11, 14,20). The
-adrenergic system has also been associated withthe memory-modulating effects of stress in humans. So far,all of the studies performed have shown that this system isparticularly involved in the enhancement of memory forevents that triggered the stressful experience in the firstplace. Indeed, pre- or postlearning stimulation of the
-adrenergic system enhances memory for the events that trig-gered the stressful experience (21–23), whereas blockade of
-adrenergic receptors before learning inhibits declarativememory for these stressful events (22, 24–26).Although the studies summarized above suggest an im-portantroleofthe
-adrenergicandcorticosteroidsystemsinmodulating human memory function in a stressful context,there are still discrepancies in the potential implication ofthese systems that necessitate further investigation. First, incontrast to the well-described effects of the
-adrenergicsystem in modulating memory for events that induced astressful experience, it is unclear at this point whether thissystem is also implicated in the modulation of human mem-ory for events that are memorized after stress and are con-sequently not related to the stressful event itself. Second, inmost of the human studies measuring the effects of stress onsubsequentmemoryprocessing[withtheexceptionofCahill
et al.
(6), and Wolf
et al.
(18)], memory performance is testedonly shortly after learning (9–11, 19; for a review, see Refs.14, 20). Declarative memory formation, however, is a time-dependent process in which both short-term memory (STM)and long-term memory (LTM) processes are initiated afterlearning to consolidate new memories. The STM trace (
i.e.
First Published Online December 7, 2004
Abbreviations: AM, Morning; GR, glucocorticoid receptor; LTM,long-termmemory;MR,mineralocorticoidreceptor;PM,afternoon;p.o.,oral; STM, short-term memory; TSST, Trier Social Stress Test.
 JCEM is published monthly by The Endocrine Society (http://www.endo-society.org), the foremost professional society serving the en-docrine community.
0021-972X/05/$15.00/0 The Journal of Clinical Endocrinology & Metabolism 90(3):16971704
 Printed in U.S.A.
Copyright © 2005 by The Endocrine Societydoi: 10.1210/jc.2004-0009
1697
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memory performance tested within the first 3 h after learn-ing) grows rapidly and then decays within hours, whereasthe LTM trace (
i.e.
memory performance tested at least 3 hafter learning) grows more slowly and is relatively perma-nent(1,27).Boththe
-adrenergicandcorticosteroidsystemshavebeenshowntomodulateshort-(
e.g.
Refs.24and28)andlong-term (
e.g.
Refs. 21–26, 28; see Ref. 5) memory consoli-dation of the stressful events themselves. However, the roleplayed by these two systems in modulating the effects of astressfulexperienceonshort-andlong-termconsolidationofmaterial unrelated to the stressor is to be further defined.Accordingly, the goal of this study was to disentangle andspecifytheroleplayedbythe
-adrenergicandcorticosteroidhormonal systems in modulating short- and long-term de-clarative memory of neutral material learned after stress inhumans. Young men were administered either a blocker ofperipheral and central
-adrenergic receptors (propranolol)or an inhibitor of corticosteroid synthesis (metyrapone) be-fore being exposed to a psychological stress task. Poststressshort-term (5-min delayed recall) and long-term (1-wk de-layed recall) declarative memory was compared with post-stress short- and long-term declarative memory measuredunder a placebo condition.
Subjects and Methods
 Experimental subjects
Forty-two healthy English- and French-speaking men participated inthis experiment. The study was conducted according to human researchguidelines and was approved by the Douglas Hospital Research EthicsBoard. Informed consent was obtained from all subjects, who werecompensatedfortakingpartinthestudy.Subjects,agedbetween18and34 yr, were randomly assigned to one of three experimental conditions:oral (p.o.) placebo (n
14); propranolol 80 mg p.o., a blocker of pe-ripheral and central
-adrenergic receptors (n
14); or metyrapone, aninhibitor of corticosteroid synthesis (two doses, 750 mg p.o. each; n
14).Age,bodymassindex,andeducationlevelwerenotdifferentamonggroups(
P
0.27).Allsubjectswerenonsmokers.Subjectswererecruitedin the community and underwent psychological and physical exami-nations as well as blood and urine laboratory routine tests. To be in-cluded in the study, individuals were required to be exempt of a presentor lifetime history of psychiatric disorders (axis I and II disorders) asevaluated with the Structured Clinical Interview for Diagnostic andStatistical Manual of Mental Disorders-IV (29). Participants were alsorequired to be medication free and exempt of current medical problems,clinically significant electrocardiogram abnormalities, and any signifi-cant abnormalities on laboratory tests. Individuals working night shiftswere excluded from the study. Care was also taken not to evaluatesubjectsduringstressfulperiods(suchasexamperiods),andindividualswho had undergone major life changes (
e.g.
death of a close familymember in the past year) were not included in the study. Women wereexcluded from the study to avoid any confounding effects of the men-strualcycleandcontraceptivepillondeclarativememory(30),totalbasalcorticosteroid levels (31), and
-blocker catabolism (32).
 Declarative memory task
Subjectsvieweda5-minnarratedseriesof11colorpicturespresentinga neutral story. Specifically, a young girl engaged in a safely organizedwoodworking activity with her grandfather before going, later on, to anoptometrist appointment with her grandmother. Narratives accompa-nying each picture were neutral and matched with regard to the com-prehensibility, complexity, syntax, and grammatical structure.Given the incidental nature of the declarative memory task (
i.e.
sub- jects were not aware of the later memory evaluation; Ref. 33), subjectswere told that they would be viewing pleasant, unpleasant, and neutralpictures and that we were interested in their physiological reactions (
i.e.
heart rate levels, blood pressure levels, and hormone levels) to thesepictures. Participants were asked to relax and simply watch the storypresented as if they were at the movies. Free recall of the story wasassessed at two time points, namely, 5 min after viewing (
i.e.
during thefirst experimental session) and 1 wk later (
i.e.
during the second exper-imental session; see
Psychoneuroendocrine protocol
). At the end of the firstsession,participantsweregivenaphonecallmeeting,tooccur1wklater,to complete a questionnaire on emotions. When called back, subjectswere informed that no questionnaire needed to be completed and wereinstead asked to recall as much information as possible about the storyviewed 1 wk earlier (long-term declarative memory condition). At theend of the phone call meeting, the experimenter asked the subjectswhether they anticipated the declarative memory tests; all reported thatthe declarative memory tests were unsuspected, for both the short- andlong-term free recall. All subjects were debriefed with respect to the realgoal of the study at the end of the second session.For the short- and long-term recall conditions, participants wereencouragedtorememberasmuchastheycouldaboutthemainstorylineas well as any details that would come to mind. Free recall was taperecorded to be analyzed later. The experimenter scoring the results wasunaware of drug conditions. Subjects were credited with the recall of apicture(foratotalofonepointperpicture)iftheyrememberedelementsthat could only have been seen in that particular picture and not in anyother picture or mentioned in the narration. The total declarative mem-ory task procedure, including the viewing of the story and the 5-mindelayedrecall,took15mintobecompleted.FrenchandEnglishversionsof the story were used, and there were no differences in short- andlong-term free recall with regard to the language in which the story waspresented (
P
0.10).
 Psychological stress protocol
Subjectsweresubmittedtoapsychologicalstresstask,theTrierSocialStress Test (TSST), developed by Kirschbaum
et al.
(34). This laboratorystressorconsistsofafreespeechandamentalarithmetictaskperformedin front of an audience. The total procedure, including preliminaryinstructionsandapreparationperiod,takes20min.Awealthoffindingsdemonstrated the efficacy of the TSST in substantially increasing freesalivary cortisol levels and cardiac activity (
e.g.
Refs. 9–11, 18, 34, 35).
 Psychoneuroendocrine protocol
Subjects were tested individually on two separate occasions. For thefirstexperimentalsession,participants wereaskedtohavealightbreak-fast at no later than 0700 h, during which they were asked not to haveany citrus products, coffee, tea, and sweets. They were also asked toavoid exercising the morning of the initial session and were asked nottoeatanddrinkanythingbutwater1hbeforethestartoftheexperiment.Subjects arrived at the laboratory at 1130 h, and baseline free salivarycortisol samples, heart rate, and blood pressure measures were taken at1150 and 1200 h. Cardiac activity was measured with the Welch AllynAtlas vital signs monitor (Skaneateles Falls, NY). At 1205 h, a first doseof metyrapone 750 mg p.o. was administered, whereas the placebo andpropranolol groups were given placebo pills. A light lunch was givenat 1245 h, and additional saliva samples, heart rate, and blood pressuremeasures were taken at 1225, 1315, 1340, and 1420 h.At 1450 h, heart rate and blood pressure measures were taken andthen, at 1452 h, the placebo group received a placebo, whereas themedication groups received their respective drugs (propranolol 80 mgp.o. or second dose of metyrapone 750 mg p.o.). These specific drugdoses (80 mg propranolol, 2
750 mg metyrapone) were selected be-cause they had proven efficient in blocking peripheral and central
-adrenergic receptors (24, 36) and inhibiting corticosteroid secretion (24,37–40) in humans. No side effects (nausea, dizziness, fatigue, sedation,
etc.
) due to either propranolol (80 mg) or metyrapone (2
750 mg) werereported by subjects. The times of drug administration were also care-fullyselectedtoensurepeakplasmapropranololandmetyraponeeffectsat the time of declarative memory evaluation (24, 37–41). The subjectsand experimenter were unaware of drug conditions.Saliva samples, heart rate, and blood pressure measures were takenat 1520 and 1550 h, and subjects were submitted to the TSST at 1555 h.Additionalsalivasamples,heartrate,andbloodpressuremeasureswerethen taken every 10 min from 1605 to 1705 h. Five minutes after the
1698
J Clin Endocrinol Metab, March 2005, 90(3):16971704 Maheu
et al.
Modulation of Declarative Memory after Stress
 by on June 8, 2009 jcem.endojournals.orgDownloaded from 
 
psychological stress task ended (
i.e.
at 1620 h), subjects viewed theneutral story and were submitted to a surprise free recall 5 min later (
i.e.
at 1630 h). Subjects left the laboratory at 1715 h, after being examined bythe medical supervisor of the study (R.J.). The second experimentalsession occurred 1 wk later, at which time participants were called overthephoneandaskedtorecallthestory(seeearlier
Declarativememorytask
description).
 Salivary cortisol assays
Free salivary cortisol samples were collected with the Sarstedtsalivette device (Sarstedt, Germany) and stored at
20 C until assayed.Samples were thawed and spun at 3000 rpm and 4 C for 20 min, andcortisol concentrations were determined by RIA with a kit from Diag-nostic Systems Laboratories (Webster, TX). Salivary samples of cortisolwere mixed with 500
l
125
I-labeled corticosteroid reagent and 500
lcortisolantiserumcomplexreagent.Totalbindingandnonspecificbind-ing typically ranged from 47 to 63 and 0.5 to 1.5%, respectively. Boundantigens were separated through a prereacted double-antibody system.When this technique is used, cross-reactivity of the antigen is less than4% with 11-deoxycortisol and less than 1% with any other naturallyoccurring steroids. The intra- and interassay coefficients of variationwere 4.6 and 5%, respectively. The limit of detection of the assay was0.01
g/dl. All samples were assayed in duplicates.
 Statistical analyses
Because of data lost for salivary cortisol and cardiac activity mea-sures, one measure of salivary cortisol, heart rate, and systolic anddiastolic blood pressure was estimated for one subject according to theformulausedbyCochranandCox(42).Physiologicalandcognitivedatawere verified for assumptions of normality and sphericity, and loga-rithmic transformations or Greenhousse-Geiser corrections (43) wereapplied when normality or sphericity was not met. Although the sali-vary cortisol data were logged to allow proper statistical analyses, sal-ivary cortisol results are presented as untransformed results in micro-grams per deciliter units for the sake of comparison between studies.Mixed ANOVAs using treatment (placebo
vs.
propranolol
vs.
me-tyrapone) as the between-subjects factor and salivary cortisol samples,heart rate, and systolic and diastolic blood pressure measures as thewithin-subjects factors were performed to evaluate the effects of treat-ment on physiological measures. A mixed ANOVA using treatment asthebetween-subjectsfactor(placebo
vs.
propranolol
vs.
metyrapone)andtime of recall (short- and long-term free recall) as the within-subjectsfactor was performed to measure the effects of treatment on the meanpercentage recall of material. Simple effects and, when appropriate,Student Newman-Keuls
post hoc
tests, were conducted on all significantphysiological and cognitive findings.
Results
 Effects of treatment on salivary cortisol measures
The ANOVA measuring the effects of treatment on sali-vary cortisol samples revealed that there was a significanttwo-way interaction between treatment and salivary cortisolsamples[F(12,224)
5.25,
P
0.01].Forfreesalivarycortisolmeasures taken before the TSST,
post hoc
comparisons between groups demonstrated group differences at 1340 h(Fig. 1). At that time, the placebo and propranolol groupswere under the influence of placebo pills (because propran-olol 80 mg was only administered after cardiac parameterswere measured, at 1452 h; see
Subjects and Methods
), whereasthe metyrapone group was under the influence of the firstdose of metyrapone (750 mg p.o.; see
Subjects and Methods
).Results showed that salivary cortisol levels in the placeboand propranolol groups were significantly increased, com-pared with those in the metyrapone group (
P
0.01). Theseresults are attributable to the fact that metyrapone had al-ready started to decrease salivary cortisol levels in the me-tyrapone group at 1340 h, whereas salivary cortisol levelswereenhancedintheplaceboandpropranololgroupsatthattime due to the administration of a light lunch at 1245 h (37,44, 45).For free salivary cortisol measures taken during and afterthe TSST,
post hoc
comparisons between groups showed thatthe placebo and propranolol groups presented significantstress-induced elevations in salivary cortisol levels, whereascortisol reactivity to the stressor was significantly sup-pressed by the metyrapone treatment (
P
0.005; Fig. 1).Finally,
 post hoc
analysesrevealedthatsalivarycortisollevelsin the propranolol group remained significantly elevated atthe end of the study (the 1705-h sample), compared withsalivary cortisol levels in the placebo and the metyraponegroups (
P
0.03; Fig. 1). Salivary cortisol levels in the pro-pranolol group, therefore, were returning to baseline levelsmore slowly than salivary cortisol levels in the placebogroup. This result is consistent with previous human find-ings showing increased cortisol levels after propranolol ad-ministration (24, 46–49).
F
IG
. 1. Mean(
SEM
)salivarycortisollevelsinthe placebo, propranolol, and metyraponegroups.
*
, Placebo and propranolol groupspresent significantly higher salivary cortisollevels than metyrapone group (
 P
0.01) and
**
,
P
0.005.
***
, Propranolol group presentssignificantly higher salivary cortisol levelsthan placebo and metyrapone groups (
 P
0.03).
Maheu
et al.
Modulation of Declarative Memory after Stress J Clin Endocrinol Metab, March 2005, 90(3):16971704
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