Behavioural Brain Research 136 (2002) 349 /357 www.elsevier.

com/locate/bbr

Research report

Effect of tryptophan depletion on impulsive behavior in men with or without a family history of alcoholism
John Crean, Jerry B. Richards 1, Harriet de Wit *
Department of Psychiatry, University of Chicago, 5841 S. Maryland Avenue, Chicago, IL 60637, USA Received 2 January 2002; received in revised form 12 March 2002; accepted 10 May 2002

Abstract This study investigated the effects of acute serotonin depletion on two measures of impulsive behavior in healthy men with a family history of alcoholism. Serotonin has been implicated in several forms of impulsive behavior, as well as in the etiology of Type II alcoholism. The present study was designed to determine if an acute disturbance of serotonin function would increase impulsive responding on two behavioral indices of impulsivity, and whether this effect would be greater in individuals with a genetic predisposition to alcoholism. Forty healthy men, half of whom had an alcoholic father, participated in a two-session study. Subjects ingested a tryptophan-depleting diet on one session and a balanced diet on the other session, and completed tasks measuring behavioral inhibition and delay discounting. Tryptophan depletion impaired performance on the behavioral inhibition task in the males with a positive family history, relative to the males without alcoholic relatives, whereas it improved behavioral inhibition in the family history negative group. Tryptophan depletion had negligible effects on mood, and it did not alter performance on the delay discounting task. The results provide partial support for the hypothesis that impulsive behavior is related to low serotonin function, and further suggests that the role of serotonin depends on genetic factors related to alcoholism. The results complement the results of a parallel study investigating the effects of serotonin depletion on a similar behavioral inhibition procedure in rats. Parallel studies in rats and humans are important to validate the large body of neurobiological research with non-human species to humans. # 2002 Elsevier Science B.V. All rights reserved.
Keywords: Impulsivity; Tryptophan depletion; Stop Task; Family history alcoholism

1. Introduction Type II alcoholism has been linked to a disturbance of the central serotonin (5-HT) system. Compared with non-alcoholics and other alcoholics, type II alcoholics tend to have significantly lower 5-HT concentrations in the cerebrospinal fluid [25,47,60,61]. Interestingly, even sons of type II alcoholics who do not develop alcoholism exhibit deficient 5-HT functioning [15,42], suggesting that 5-HT represents one of the systems involved in the genetic abnormality underlying this syndrome. Type II alcoholism occurs primarily in males, begins at an early age, has a strong heritable component and is

* Corresponding author. Tel.: /1-773-702-1537; fax: /1-773-8347698 E-mail address: hdew@midway.uchicago.edu (H. de Wit). 1 Present address: University of New York at Buffalo.

associated with a high incidence of impulsive aggression and anti-social personality traits [8,9]. The present study was designed to examine the role of serotonin in risk for alcoholism and susceptibility to impulsive behavior. Most of the clinical research linking serotonin to type II alcoholism has been correlational. However, studies with laboratory animals provide some support for the idea that excessive alcohol consumption is related to a 5HT-deficit. Depleting brain levels of 5-HT in rodents, for example, increases both alcohol consumption [36] and impulsive aggression [58,59], and these effects can be reversed by restoring 5-HT function. Alcohol-preferring rodent strains tend to have lower 5-HT levels than non-preferring rodent strains [35], and lower 5-HT levels are associated with heightened aggression and increased alcohol consumption in non-human primates [20]. Finally, 5-HT depleted animals exhibit impaired performance on operant tasks that require rapid inhibi-

0166-4328/02/$ - see front matter # 2002 Elsevier Science B.V. All rights reserved. PII: S 0 1 6 6 - 4 3 2 8 ( 0 2 ) 0 0 1 3 2 - 8

350

J. Crean et al. / Behavioural Brain Research 136 (2002) 349 /357

tion of prepotent or dominant appetitive responses [18,56]. Researchers have proposed several different behavioral processes that may mediate these patterns of behavior, including impairments in decision-making [45] or impairments in response inhibition [21,57]. The findings in laboratory animals need to be extended to humans to establish the link between 5HT and alcoholism. One way to study the role of 5-HT in humans is to temporarily reduce brain levels of 5-HT through a tryptophan depletion diet [17,22,43]. Acute tryptophan depletion has been reported to increase aggression on a standardized behavioral task in healthy adult volunteers [7,33], and to impair behavioral inhibition in adult non-alcoholic men with a history of multigenerational paternal alcoholism [23]. The aim of this experiment was to examine the relationship between 5-HT and impulsiveness in males at risk for alcoholism. Two groups of young adult, nonalcoholic men participated in a tryptophan depletion procedure. One group (family history positive, FHP) had fathers with type II alcoholism and the other group (family history negative, FHN) had no family history of alcoholism. Subjects participated in two sessions in which they received either a tryptophan-depleting amino-acid beverage (TD) or a tryptophan-balanced beverage (TB), before performing two tasks to assess impulsivity. They completed the Stop Task [27], which assesses the capacity to inhibit a pre-potent or ongoing response, and a delayed reward discounting procedure [45], which estimates the rate at which a monetary reward is devalued across a series of delays. Both tasks are considered to be sensitive, reliable and valid measures of forms of impulsive behavior [11,45,46,50 / 52]. It was hypothesized that the subjects would perform more impulsively on the tasks after tryptophan depletion and that this effect would be more pronounced in the FHP group.

proved by the University of Chicago Medical Center Institutional Review Board. 2.2. Recruitment and screening Participants were recruited through advertisements placed in community newspapers and flyers posted at local colleges and universities. At the screening interview, prospective participants completed the Michigan Alcohol Screening Test (MAST; [54]) and the Psychiatric Symptom Checklist (SCL-90; [14]) to screen for alcohol and psychiatric problems. A clinician interviewed the subjects about current and past medical conditions, recreational drug use, and psychiatric symptoms. Individuals with a current or a past history of serious medical conditions, or with current or past psychiatric illness (DSM-IV Axis I psychiatric disorder), including substance use disorders except alcohol abuse, were not accepted. An adapted form of the Family History Research Diagnostic Criteria (FH-RDC; [2]) was used to assess family history of substance use and psychiatric disorders. To qualify for the FHP group subjects were required to have fathers who met FH-RDC criteria for Type II alcohol dependence with at least two of the following features: an onset before age 25, frequent fights while drinking; driving under the influence of alcohol or alcohol-related motor vehicle accidents; and arrests under the influence of alcohol. Participants in the FNH group had no history of substance abuse/dependence in their immediate family members and firstdegree relatives. Participants attended an orientation session during which they signed the consent form and completed personality questionnaires (see below). 2.3. Laboratory sessions Each subject participated in two 9-h laboratory sessions, conducted at the University of Chicago Clinical Research Center. The sessions were conducted from 7:30 a.m. to 4:30 p.m., at least 5 days apart. Participants fasted starting at midnight the night before the sessions, and used no drugs for 24 h before the session (including alcohol, marijuana, aspirin, or other drugs, or more than 3 cups of coffee, tea, cola or more than 6 cigarettes per day). Urine and breath samples were obtained to verify compliance (no tests were positive). After the urine screen, a nurse inserted an intravenous catheter into the participants non-dominant forearm and drew 10 cc of blood for baseline determination of tryptophan levels. The amino acid (AA) mixture (described below) was administered immediately after the first blood draw, at 8 a.m. Additional blood samples were obtained 2, 4, 6 and 8 h post AA consumption to document the effect of the dietary manipulations on

2. Methods 2.1. Participants Forty healthy males, aged 18 /25, who were fluent in English and had at least a high school education completed this study. The participants were estimated to be in the above-average range of intellectual functioning (National Adult Reading Test, NART [11]). They were medically and psychiatrically healthy, and most reported light recreational drug use (Table 2). The participants were assigned to the FHP group if their fathers met criteria for Type II alcoholism (see below), and to the FHN group if they had no immediate family members or first-degree relatives with a history of substance abuse or dependence. This study was ap-

J. Crean et al. / Behavioural Brain Research 136 (2002) 349 /357

351

plasma tryptophan level. Nurses separated plasma from whole blood using centrifugation and froze the samples immediately. Total plasma tryptophan concentrations were measured at a later date with high performance liquid chromatography with fluorometric detection [1]. Participants completed a baseline version of the Stop Task immediately after consuming the AA mixture, before the dietary manipulation could affect serotonin levels. They performed both the Stop Task and delay discounting procedure at 1:00 p.m., 5 h after administration of the diet, at a time when tryptophan levels in the active condition are expected to be low [6]. The POMS was administered every hour during the session to monitor the effects of the diet on mood. At times when no experimental events were scheduled, participants were allowed to relax in their rooms and read or watch television. They were not allowed to eat until 2:30 p.m., but were allowed to drink water. At 2:30 p.m. they were given a small snack, and the sessions ended at 4:30 p.m. After completing both sessions subjects attended a debriefing interview and received payment.

2.4. Amino acid mixtures A licensed nutritionist prepared the AA mixtures following the method described by [62]. Briefly, on the balanced session (TB) subjects received all of the sixteen AAs in ratios equivalent to those found in commonly consumed proteins. On the tryptophan depletion session (TD) subjects received same mixture except that the tryptophan was deleted. The TB and TD mixtures were prepared identically. Most of the AAs were administered in a solution of ice slurry with small amounts of chocolate syrup and saccharine added for flavor. The remaining AAs were put in capsules (fourteen size 0 capsules) because of their unpleasant taste. The beverages and capsules were administered under double blind conditions.

2.5.2. Stop Task [27] The Stop Task measures the ability to inhibit prepotent behaviors [4], based on a theoretical model of behavioral inhibition [26,28]. Children with ADHD perform more poorly on this task, and their performance is improved with administration of the clinically effective drug methylphenidate [57]. The task is based on a mathematical model known as the race model in which there is a competition between processes involved in executing and inhibiting responses. On repeated trials in the task, a Go signal is presented on the computer screen, and subjects are instructed to respond as quickly as possible when this signal appears. The Go signal appears at 2-s intervals, and the individuals reaction times (Go RT) are recorded. On 25% of the trials, a tone (Stop signal) is sounded very shortly after the presentation of the Go signal, and participants are instructed to inhibit (stop) their response. The delay from the onset of the Go signal to the onset of the Stop signal (stop signal delay) is systematically increased or decreased in 50 ms increments, until the subject is able to inhibit (stop) his or her key press responses on approximately 50% of trials. Stop RT is computed by subtracting the average stop signal delay at which the individuals are able to inhibit their Go response 50% of the time from the average Go RT. The Stop Task was presented in 4 blocks of 64 trials (256 total trials), with a brief rest period between each block of 64 trials. It took subjects about 10 min to complete all 256 trials. At the beginning of the task the stop delay was set at 200 ms. Because the data on the first blocks were variable as the Stop RT was determined, only the data from the last block of 64 trials were used in the analysis. The procedure provides a measure of (i) the latency in milliseconds to respond to the letter presentation, or the Go RT, (ii) the accuracy in responding appropriately to the Go signal, and (iii) the percentage of trials on which the individual fails to inhibit the response when the Stop signal is presented, (iv) the time in ms needed to inhibit a Go response (Stop RT). The Stop RT is the primary outcome measure. 2.5.3. Delay discounting task [44] The delay discounting task measures preference for immediate, smaller rewards over larger, more delayed rewards [44]. It is based on a definition of impulsivity positing that impulsive individuals are more sensitive to immediate, as opposed to delayed consequences, and therefore they value immediate rewards more than delayed rewards [3,19,29,41]. The delay discounting task [44] uses a computerized adjusting amount procedure to measure discounting of delayed reinforcers. Subjects have the opportunity to choose between different amounts of money available after different delays. The test consists of about 110 questions, such as: Would you rather have $10.00 in 30 days or $2.00 at the

2.5. Dependent measures

2.5.1. Prole of mood states POMS; [32] The POMS is a 72-item rating scale that is sensitive to momentary changes in mood state. It consists of 72 adjectives commonly used to describe mood. Participants rate the extent to which each adjective describes how they feel at the moment on a scale from (0) not at all to (4) extremely. The POMS is comprised of eight factor-analyzed primary scales including (1) Depression; (2) Vigor; (3) Elation; (4) Anxiety; (5) Fatigue; (6) Confusion; (7) Arousal and (8) Friendliness. Subjects scores on these 8 scales were analyzed. The POMS was completed every hour during the sessions.

352

J. Crean et al. / Behavioural Brain Research 136 (2002) 349 /357

end of the session? The questions are presented on a computer screen according an adjusting amount procedure [44], in which the amount of immediate, certain money is adjusted across successive questions (trials) until an amount is reached that is judged by the participant to be equivalent to the delayed $10.00 reward. The amount of immediate, certain money the subject judges to be equivalent to the delayed $10.00 reward provides a quantitative measure of the subjective value of the delayed rewards. This point of subjective equality is called indifference points. Within each session, indifference points are determined for five different delays, 0, 2, 30, 180, and 365 days. On each trial, participants choose between a varying amount of money available immediately and $10 available after one of these delays. At the end of the session, when all questions have been answered, one question/response is selected at random and the subject receives whatever they chose in response to that question (i.e. money available immediately or after the designated delay). The indifference points obtained at each of the delays are then plotted to form a discount function. The delay discounting function is derived through curve-fitting analyses, which yields a value for the parameter k . The curves that result from the devaluation of reward value by delay are well described by the hyperbolic function of Mazur [30], V /A /(1/kD ), where V is value, A is the amount of the delayed reward, D is the delay to reward, and k is a free parameter. Larger values of k indicate more rapid devaluation of reinforcer value by delay and greater impulsivity. 2.5.4. Barratt Impulsiveness Scale (BIS11; [39]) The BIS is a questionnaire designed to measure the personality trait of impulsivity. It consists of 30 items on which subjects describe themselves, from never to almost always. The BIS11 has been factor analyzed into 3 subscales, corresponding to motor impulsivity, attentional, or cognitive impulsiveness and non-planning impulsiveness. The BIS11 distinguishes samples known to be impulsive (i.e. substance abuse patients, general psychiatric patients and prison inmates) from control subjects (undergraduates) [39]. 2.6. Data analysis The demographic characteristics of the two groups were analyzed using independent samples t -tests for continuous variables and chi square tests for categorical variables. In the delay discounting task, the hyperbolic discount function was fit to the five delay indifference points obtained from each participant using a nonlinear curve fitting program [37]. This curve-fitting program determined the best fitting values for k and the coefficient of determination (r2). Dependent measures, including total plasma tryptophan concentrations, go

and stop reaction time, delay discounting data and POMS factor scores, were analyzed using separate mixed-plot analyses of variance (ANOVA), using Huynh-Feldt Epsilon to correct for sphericity [21]. For the Stop Task (Go and Stop RTs) the factors in the ANOVAs were Condition (TB and TD) and Group (between-subjects: FHN and FHP). For the plasma tryptophan and POMS data the factors were Condition (TB and TD), Group (FHN and FHP) and time (baseline, 2, 4, 6 and 8 h). For the delay discounting task the factors were Condition (TD and TB) /Group (FHN and FHP) /Delay (0, 2, 30, 180 and 365 days). Statistical significance was set at the 0.05 level, and in cases of multiple tests on similar data, the alpha level was adjusted to correct for increased probability of Type I error (i.e. Bonferoni correction).

3. Results 3.1. Group characteristics The FHP (n /20) and FHN (n /20) groups did not differ in age, education, ethnicity or estimated intelligence (NART; Table 1). Most subjects were either in college or had recently graduated from college. The groups did not differ significantly on any of the BIS subscales (cognitive, motor, non-planning). Most of the participants reported occasional use of illicit drugs, and about half of the subjects in both groups reported smoking tobacco cigarettes daily (Table 2). All of the participants reported consuming alcohol, but the FHP participants reported heavier weekly alcohol use than
Table 1 Demographic and drug use data for FHP and FHN groups Characteristic FHP (n  20) 22.392.1 14.592.9 115.595.6 68.5913.7 15/3/1/1/0 FHN (n  20) 21.591.8 14.691.4 114.196.3 63.599.1 12/1/2/2/3

M M M M

age (years) education (years) estimated IQ BIS total score

Ethnicity Caucasian/Af. Amer./Hisp/Asian/SE Asian (Indian) Substance use history M MAST score M # of weekly alcohol beverages % cigarette smokers (  7/day) % with  50 lifetime cocaine/stimulant use % with  50 lifetime heroin/opiate use % with  50 lifetime marijuana use

11.8910* 10.997* 55% 10% 0 80%

2.5592 6.993 45% 0 0 50%

Scores shown are mean values (M ), with standard error, frequency (ethnicity) or percent of subjects who used a drug. Asterisk indicates significant difference between the groups.

J. Crean et al. / Behavioural Brain Research 136 (2002) 349 /357 Table 2 Percent of FHP fathers who met each of the features of Type II alcoholism Type II feature Onset B 25 One or more alcohol abuse treatment attempts Frequent driving while drunk Frequent fights while intoxicated One or more arrests Evidence of polysubstance abuse Anti social traits n  20 100% 75% 95% 95% 75% 20% 35%

353

the FHN controls. In addition, the FHP group scored higher on the MAST and four of the FHP participants met criteria for DSM-IV alcohol abuse, whereas none of the FHN participants met these criteria. 3.2. Family history characteristics Half of the subjects in the FHP sample reported multigenerational family history of alcoholism in addition to their fathers, and one FHP participant also reported a history of maternal alcoholism. Table 2 lists the percent of FHP fathers with symptoms consistent with the various type II clinical features. The FH-RDC provided an estimate of the incidence of psychiatric disorders in the participants first-degree relatives. Twenty-five percent of the relatives of the FHP subjects, and 15% of the relatives of FHN subjects had at least one clinically significant psychiatric disorder in their lifetime. Disorders among FHP subjects relatives included 4 cases of major depression and 1 case of panic disorder, and the FHP subjects had 3 relatives with major depression. 3.3. Primary dependent measures 3.3.1. Plasma tryptophan level As expected, total plasma tryptophan levels were lower following the TD manipulation (main effect of condition, F /232.04 (2, 68), P B/0.001; Table 3). The groups did not differ in total plasma tryptophan levels either before or after the diets. The TB diet increased plasma levels of tryptophan.

3.3.2. Stop Task Separate ANOVAs were used to analyze Go RT (primary reaction time) and Stop RT. For Go RT, there were no differences between the groups, no effects of the tryptophan depletion, and no interactions (Fig. 1). For the Stop RT, there was a significant condition /group interaction (F /8.78 (1, 38) P B/0.01). On this measure tryptophan had opposite effects in the FHP and FHN groups. Post-hoc tests revealed that the FHP group exhibited longer Stop RTs compared to the FHN group after tryptophan depletion, but the groups did not differ in the TB condition (Fig. 1). Examination of the means showed that Stop RTs increased after TD in FHP participants whereas Stop RTs decreased after TD among the FHN participants.

3.3.3. Delay discounting On the delay discounting task there was a main effect of Delay (F /104.50 (4, 148) P B/0.001), but there were no effects of group (FHN and FHP) or tryptophan depletion (Fig. 2). When hyperbolic functions were fit to the points there were no significant differences in k values among any of the four conditions (FHP, FHN, TD, TB). The mean (and sem) k values for the TB condition were 0.017 (0.005) for the FHN group and 0.59 (0.31) for the FHP group. The mean k values for the TD condition were 0.029 (0.012) for the FHN group and 0.58 (0.30) for the FHP group. The r2 values were between 0.74 and 0.81.

Fig. 1. Mean (s.e.m.) Go reaction time and Stop reaction time in ms on the Stop Task, in FHP and FHN groups after the balanced diet (TB) and the tryptophan depletion diet (TD). Go reaction time was not different across the four conditions. In the TD condition, the FHP group exhibited longer Stop reaction times than the FHN group.

Table 3 Mean (and S.D.) plasma levels of tryptophan (mg/ml) after balanced and depletion diets in FHP and FHN subjects Pre FHP balanced FHP depleted FHN balanced FHN depleted 11.692.7 10.792.3 13.493.0 13.593.2 2h 34.3911.2 3.791.4 38.2911.3 5.292.7 4h 23.296.9 1.490.7 26.399.1 1.992.4 6h 14.595.3 2.190.9 14.995.1 2.292.2 8h 10.593.1 2.892.7 10.993.1 2.892.2

354

J. Crean et al. / Behavioural Brain Research 136 (2002) 349 /357

Fig. 2. Median indifference points (and tted curve) in the FHP and FHN groups after the balanced diet (TB) and the tryptophan depletion diet (TD). There were no signicant differences between the groups or across the tryptophan conditions.

3.3.4. POMS Tryptophan depletion diet had little effect on mood. Using data from the baseline and the measure obtained at 5 h after the diet (when tryptophan depletion was expected to be maximal and when performance measures were obtained), there was only one significant interaction between group and tryptophan depletion. Ratings on the Anxiety scale declined after TD in the FHN group but not in the FHP group (Anxiety: Group /Treatment interaction, P B/0.05, df /1.42, F /6.19). There were no systematic relationships between mood and performance on the impulsivity tasks.

4. Discussion This study examined the effects of tryptophan depletion on two behavioral measures of impulsiveness in non-alcoholic young men with or without histories of paternal type II alcoholism. It was hypothesized that tryptophan depletion would increase impulsivity, particularly in FHP individuals. The results provided partial support for the hypothesis: On the Stop Task, a measure of response inhibition, the FHP group performed more poorly (i.e. more impulsively) after tryptophan depletion than the FHN group. However, on the measure of delay discounting, there were no differences between the groups and no effects of tryptophan depletion. Assays of plasma tryptophan levels confirmed that the depletion procedure effectively reduced tryptophan level. Self-report measures indicated that the tryptophan depletion procedure had minimal effects on mood. These findings suggest that individuals with a family history of alcoholism may be more susceptible to behavioral impairment following a rapid decline in brain tryptophan levels, providing further support for the role of serotonin in behavioral inhibition. The data also suggest that the impairment resulting from serotonin depletion is specific to one type of impulsivity, i.e. to the ability to inhibit prepotent responses.

The present results suggest that the Stop Task and delay discounting task measure different aspects of impulsive behavior. Although both measures have some validity insofar as they differentiate populations known to differ in impulsivity [11,12,25,39,40], performance on the measures is not correlated, and in the present study, tryptophan depletion affected performance on the Stop Task but not on delay discounting. It is possible, however, that the Stop Task is simply a more sensitive index of impairment than the delay discounting procedure. Indeed, despite its validity as a measure of trait impulsivity, the human version of the delay discounting procedure may be relatively insensitive to changes in the state of impulsivity, as studied here after TD. In previous studies [46] we have failed to detect changes in delay discounting after administration of acute doses of drugs, including alcohol, that are expected to increase impulsivity. In contrast, the Stop Task has been found to be sensitive to the effects of acute doses of drugs including amphetamine and alcohol [16]. It is also possible that the Stop Task and delay discounting tasks measure different subtypes of impulsivity, and that only the type that is involved in the Stop Task is mediated by serotonin. The reductions in plasma levels of tryptophan in the present study were similar to the effects observed in other studies, indicating that the dietary manipulation was effective. In the present study, plasma tryptophan levels at baseline were 10 /14 micrograms per ml before the diet, and 2 /3 mg/ml 5 h after the depletion. These levels are comparable to what has been observed previously [24,34,48], and are likely to indicate a decrease in central serotonin turnover [6]. The TB diet increased plasma levels of tryptophan, but such increases are not thought to affect brain function because most dietary tryptophan is not metabolized to serotonin. Although tryptophan depletion sometimes induces negative mood states in individuals with depression or with a family history of depression [5,13], tryptophan depletion usually does not induce negative mood states in healthy volunteers [38,48,49,55]. Consistent with this, the procedure produced negligible changes in mood in the subjects in the present study. One unexpected finding in the present study was that TD in the FHN group decreased Stop RTs, that is, it made the subjects perform in a less impulsive manner. Although this is not consistent with the idea that low serotonin function is associated with high impulsivity, it is consistent with some other studies in which tryptophan depletion improved performance in healthy volunteers. For example, Rubinsztein et al. [48] found that tryptophan depletion improved performance on an affective set-shifting task compared to a control condition. In addition Schmitt et al. [53] reported that tryptophan depletion improved performance on tasks requiring focussed attention (Stroop test and dichotic

J. Crean et al. / Behavioural Brain Research 136 (2002) 349 /357

355

listening), and Coull et al. [10] reported a decrease in reaction times on a stimulus response incompatibility task. Rubinsztein et al. [48] suggest that these improvements in performance may be due to a reduction of affective attentional set, resulting in less interference on a complex task. Thus, in the present study it is possible that Stop RT improved in the FHN group because the TD removed an inhibitory cognitive influence normally operative during a complex task. The results of the present study can also be related to a similar study by LeMarquand et al. [23], investigating the effects of tryptophan depletion in males with or without a multigenerational family history of alcoholism on measures of aggression and disinhibition. Aggression was measured using the Taylor aggression task, and disinhibition was measure using a go-no-go task. Tryptophan depletion had no effect on the Taylor aggression task in either group. However, tryptophan depletion increased errors of commission on the go-nogo task, only in FHP subjects. These findings suggested that lowered levels of serotonin may increase impulsive, disinhibited behavior in certain vulnerable individuals. In our study TD did not significantly increase Stop RT in the FHP group (although there was a change in the expected direction). Several procedural factors may account for the differences in outcome between the LeMarquand et al. study and our current study. One is that the go-no-go procedure may measure a different aspect of impulsivity than the Stop Task, which may be more sensitive to disruption of serotonin function. In addition, the FHP subjects in their study may have had higher genetic risk because they had a multigenerational family history, whereas subjects in our study were only required to have an alcoholic father. Previous studies (e.g. [31]) suggest that the genetic risk is greater in offspring with multigenerational alcoholism histories. Other methodological differences could also have contributed to the differential results. Tryptophan depletion increased negative mood in the LeMarquand et al. study but not in the present study, suggesting that there were differences in the effectiveness of the tryptophan depletion procedure. Finally, the present study was a within group design with each subject receiving both the TB and TD diets whereas the LeMarquand et al. study used a between groups design with subjects received either the TB or the TD diet. If only the TD results were compared for the FHP and FHN groups in the present study, similar between-group differences would have been observed. It may be that the within group design used in the present study was more sensitive to the effects of tryptophan depletion. In a parallel study with rats, we (Richards et al., in press) [63] recently found that large permanent depletions of 5-HT slowed Stop RT without altering Go RT. Similarly, TD in humans also affected Stop RT and had no effect on Go RT. However, the effects of TD on Stop

RT in humans were more complex. Stop RT was slowed by TD only in the FHP group while Stop RT in the FHN group was faster after TD. An obvious explanation of the differences in results between the rat and human studies is that the 5-HT depletion in the rats was permanent, and much larger, than in the humans. Indeed, given the differences in the degree of depletion, the degree of similarity in results in the two is quite remarkable. In general, both the rat and human studies indicate that 5-HT plays a role in mediating the ability to stop or inhibit an ongoing prepotent response. In summary these results indicate that low 5-HT may increase impulsive behavior by impairing behavioral inhibition, particularly in individuals who are especially vulnerable, such as the FHP group. Further, the absence of an effect of TD on the delay discounting task indicates that impulsive behavior may involve different behavioral processes and that these other behavioral processes may not be mediated by 5-HT.

Acknowledgements This research was funded by DA09133 and supported by the University of Chicago Clinical Research Center M01RR00055.

References
[1] Anderson GM, Young JG, Cohen DJ, Schlicht KR, Patel N. Liquid-chromatographic determination of serotonin and tryptophan in whole blood and plasma. Clin Chem 1981;27:775 /6. [2] Andreasen NC, Endicott J, Spitzer RL, Winokur G. The family history method using diagnostic criteria. Arch Gen Psychiat 1977;34:1229 /35. [3] Ainslie G. Specious reward: a behavioral theory of impulsiveness and impulse control. Psychol Bull 1975;82:463 /96. [4] Barkley RA. ADHD and the nature of self-control. New York: The Guildford Press, 1997. [5] Benkelfat C, Ellenbogen MA, Dean P, Palmour RM, Young SN. Mood-lowering effect of tryptophan depletion: enhanced susceptibility in young men at risk for major affective disorders. Arch Gen Psychiat 1994;51:687 /97. [6] Carpenter LL, Anderson GM, Pelton GH, Gudin JA, Kirwin PDS, Price LH, et al. Tryptophan depletion during continuous CSF sampling in healthy human subjects. Neuropsychopharmacology 1998;19:26 /35. [7] Cleare AJ, Bond AJ. The effect of tryptophan depletion and enhancement on subjective and behavioral aggression in normal male subjects. Psychopharmacology 1995;118:72 /81. [8] Cloninger CR. Neurogenic adaptive mechanisms in alcoholism. Science 1987;236:410 /6. [9] Cloninger CR, Boham M, Sigvardsson S. Inheritance of alcohol abuse: cross-fostering analysis of adopted men. Arch Gen Psychiat 1981;38:861 /8. [10] Coull JT, Sahakian BJ, Middleton HC, Young AH, Park SB, McShance RH, et al. Differential effects of clonidine, haloperidol, diazepam, and tryptophan depletion on focused attention and attentional search. Psychopharmacology 1995;121:222 /30.

356

J. Crean et al. / Behavioural Brain Research 136 (2002) 349 /357 [31] McCaul ME, Turkkan JS, Svikis DS, Bigelow GE, Cromwell CC. Alcohol and drug use by college males as a function of family alcoholism history. Alcohol Clin Exp Res 1990;14:467 /71. [32] McNair D, Lorr M, Droppleman L. Prole of mood states. San Diego: Educational and Industrial Testing Service, 1971. [33] Moeller FG, Dougherty DM, Swann AC, Collins D, Davis C, Cherek DR. Tryptophan depletion and aggressive responding in healthy males. Psychopharmacology 1996;126:97 /103. [34] Moore P, Landolt HP, Seifritz E, Clark C, Bhatti T, Kelsoe J, et al. Clinical and physiological consequences of rapid tryptophan depletion. Neuropsychopharmacology 2000;23:601 /22. [35] Murphy JM, McBride WJ, Lumeng L. Regional brain levels of monoamines in alcohol preferring and non-preferring lines of rats. Pharmacol Biochem Behav 1982;16:145 /9. [36] Naranjo CA, Sellers EM, Lawrin MO. Modulation of ethanol intake by serotonin uptake inhibitors. J Clin Psychiat 1986;47:16 / 22. [37] Origin 4.1 (Computer Software). Northampton, MA: Microcal Software Inc; 1995. [38] Park SB, Coull JT, McShane RH, Young AH, Sahakian BJ, Robbins TW, et al. Tryptophan depletion in normal volunteers produces selective impairments in learning and memory. Neuropsychopharmacology 1994;33:575 /88. [39] Patton JH, Stanford MS, Barratt ES. Factor structure of the Barratt Impulsiveness Scale. J Clin Psychol 1995;51:768 /74. [40] Petry NM. Pathological gamblers, with and without substance use disorders, discount delayed rewards at high rates. J Abnorm Psychol 2001;110:482 /7. [41] Rachlin H, Green L. Commitment, choice, and self-control. J Exp Anal Behav 1972;17:15 /22. [42] Rausch JL, Monteiro MG, Maristela G, Schuckitt MA. Platelet Serotonin uptake in men with family history of alcoholism. Neuropsychopharmacology 1991;4:83 /6. [43] Reilly JG, McTavish SFB, Young AH. Rapid depletion of plasma tryptophan: a review of studies and experimental methodology. J Psychopharmacol 1997;11:381 /92. [44] Richards JB, Mitchell SH, de Wit H, Seiden LS. Determination of discount functions in rats with an adjusting amount procedure. J Exp Anal Behav 1997;67:353 /66. [45] Richards JB, Sabol KE, de Wit H. The effects of methamphetamine on the adjusting amount procedure, a model of impulsive behavior in rats. Psychopharmacology 1999;146:432 /9. [46] Richards JB, Zhang L, Mitchell SH, de Wit H. Delay or probability discounting in a model of impulsive behavior: effect of alcohol. J Exp Anal Behav 1999;71:121 /43. [47] Roy A, Adinoff B, DeJong J, Linnoila M. Cerebrospinal uid variables among alcoholics lack seasonal variation. Acta Psychiatr Scand 1991;84:579 /82. [48] Rubinsztein JS, Rogers RD, Reidel WJ, Mehta MA, Robbins TW, Sahakian BJ. Acute dietary tryptophan depletion impairs maintenance of affective set and delayed visual recognition in healthy volunteers. Psychopharmacology 2001;154:319 /26. [49] Saloman RM, Miller HL, Krystal JH, Heninger GR, Charney DS. Lack of behavioral effects of monoamine depletion in healthy subjects. Biol Psychiat 1997;41:58 /64. [50] Schachar R, Logan G. Are hyperactive children decient in attentional capacity? J Abnorm Child Psych 1990;18:493 /513. [51] Schachar RJ, Tannock R, Cunningham C, Corkum PV. Behavioral, situational, and temporal effects of treatment of ADHD with methylphenidate. J Am Acad Child Psy 1997;36:754 /63. [52] Schachar RS, Tannock R, Logan G. Inhibitory control, impulsiveness, and attention decit hyperactivity disorder. Clin Psychol Rev 1993;13:721 /39. [53] Schmitt J, Jorissen B, Sobczak S, van Boxtel M, Deutz N, Riedal W. Effects of tryptophan depletion on cognitive functioning. J Psychopharmacol 2000;121:222 /30.

[11] Crawford JR, Parker DM, Allan KM, Jack AM, Morrison FM. The Short NART: cross-validation, relationship to IQ and some practical considerations. Br J Clin Psychol 1991;3:223 /9. [12] Crean JP, de Wit H, Richards JB. Reward discounting as a measure of impulsive behavior in a psychiatric outpatient population. Exp Clin Psychopharm 2000:155 /62. [13] Delgado PL, Charney DS, Price LH, Aghajanian GK, Landis H, Heninger GR. Serotonin function and the mechanism of antidepressant action: reversal of anti-depressant-induced remission by rapid depletion of plasma tryptophan. Arch Gen Psychiat 1990;47:411 /8. [14] Derogatis L. SCL-90-R Manual-II. Towson, MD: Clinical Psychometric Research, 1983. [15] Devor EJ. Linkage studies in 16 St. Louis families. Present status and pursuit of an adjunct strategy. Adv Neurol 1992;58:181 /7. [16] de Wit H, Crean J, Richards JB. Effects of d -amphetamine and ethanol on a measure of behavioral inhibition in humans. Behav Neurosci 2000;114:830 /7. [17] Fernstrom JD, Wurtman RJ. Brain serotonin content: physiological regulation by plasma neutral amino acids. Science 1972;178:414 /6. [18] Harrison AA, Everitt BJ, Robbins TW. Central serotonin depletion impairs both the acquisition and performance of a symmetrically reinforced go/no-go conditional visual discrimination. Behav Brain Res 1999;100:99 /112. [19] Herrnstein RJ. Self-control as response strength. In: Bradshaw CM, Szabadi E, Lowe CF, editors. Quantication of steady-state operant behavior. Amsterdam: Elsevier, 1981:3 /20. [20] Higley JD, Mehlman PT, Poland RE, Taub DM, Vickers J, Suomi SJ, et al. CSF testosterone and 5-HIAA correlate with different types of aggressive behaviors. Biol Psychiat 1996;40:1067 /82. [21] Huynh H, Feldt LS. Estimation of the Box correction for degrees of freedom from sample data in the randomized block and splitplot designs. J Educ Stat 1976;1:69 /82. [22] Klaassen T, Riedel WJ, Deutz NEP, van Someran A, van Praag HM. Specicity of the tryptophan depletion method. Psychopharmacology 1999;141:279 /86. [23] LeMarquand DG, Benkelfat C, Pihl RO, Palmour RM, Young SN. Behavioral disinhibition induced by tryptophan depletion in nonalcoholic young men with multigenerational family histories of paternal alcoholism. Am J Psychiat 1999;156:1771 /9. [24] LeMarquand DG, Phil RO, Young SN, Tremblay RE, Seguin JR, Palmour RM, et al. Tryptophan depletion, executive functions, and disinhibition in aggressive, adolescent males. Neuropsychopharmacology 1998;19:333 /41. [25] Linnoila M, DeJong J, Virkkunen M. Family history of alcoholism in violent offenders and impulsive re setters. Arch Gen Psychiat 1989;46:613 /6. [26] Logan GD. Attention, automaticity, and the ability to stop a speeded choice response. In: Long J, Baddeley AD, editors. Attention and performance IX. Hillsdale, NJ: Lawrence Erlbaum, 1981. [27] Logan GD, Cowan WB. On the ability to inhibit thought and action: a theory of an act of control. Psychol Rev 1984;91:295 / 327. [28] Logan GD, Schachar RJ, Tannock R. Impulsivity and inhibitory control. Psychol Sci 1997;8:60 /4. [29] Logue AW. Research on self-control: an integrated framework. Behav Brain Sci 1988;11:665 /709. [30] Mazur JE. An adjusting procedure for studying delayed reinforcement. In: Commons ML, Mazur JE, Nevin JA, Rachlin H, editors. Quantitative analysis of behavior: the effects of delay and of intervening events on reinforcement value, vol. 5. Hillsdale: Lawrence Erlbaum Associates, 1987:55 /73.

J. Crean et al. / Behavioural Brain Research 136 (2002) 349 /357 [54] Selzer ML. The Michigan alcoholism screening test: the quest for a new diagnostic instrument. Am J Psychiat 1971;127:1653 /8. [55] Smith SE, Pihl RO, Young SN, Ervin FR. A test of possible cognitive and environmental inuences on the mood lowering effect of tryptophan depletion in normal males. Psychopharmacology 1987;91:451 /7. [56] Soubrie P. Reconciling the role of central serotonin neurons in human and animal behavior. Behav Brain Sci 1986;9:319 /64. [57] Tannock R, Schachar RJ, Carr RP, Chajczyk D, Logan GD. Effects of methylphenidate on inhibitory control in hyperactive children. J Abnorm Child Psych 1989;17:473 /91. [58] Vergnes M, Kempf E. Tryptophan deprivation: effects of mouse killing in the rat. Pharm Biochem Behav 1981;14:19 /23.

357

[59] Vergnes M, Depaulis A, Boeher A. Parachlorophenylalanineinduced serotonin depletion increases offensive but not defensive aggression in male rats. Physiol Behav 1986;36:653 /8. [60] Virkkunen M, Linnoila M. Brain serotonin, type 2 alcoholism and impulsive violence. J Stud Alcohol 1993;11:163 /9. [61] Virkkunen M, Eggert M, Rawlings R, Linnoila M. Aprospective follow-up study of alcoholic violent offenders and re setters. Arch Gen Psychiat 1996;53:523 /9. [62] Young SN, Smith SE, Pihl RO, Ervin FR. Tryptophan depletion causes a rapid lowering of mood in normal males. Psychopharmacology 1985;87:173 /7. [63] Richards, J.B., T. Kline, D.B. Miller, J.P. Crean, H. de Wit. Reduction of brain serotonin impairs response inhibition in rats. Physiology and Behavior (in press).