Altered neuronal nitric oxide synthase expression contributes to disease progression in Huntington's disease transgenic mice.

Reduced neuronal NOS (nNOS) expression and biochemical activity was found in the striatum (P<0.05) and cerebellum P<0.05) of late-stage R6/1 Huntington's disease (HD) mice. The changes in NOS biochemical activity correlated with body weight (P<0.001), abnormal clasping (P<0.05) and motor functioning (P<0.05) scores. HD transgenic mice missing both copies of the nNOS gene showed accelerated disease progression relative to HD transgenic mice wildtype or heterozygous for the nNOS gene. On the other hand, mice with one copy of the nNOS gene had delayed onset of their HD-related symptoms relative to HD transgenic mice wildtype for nNOS. Administration of an iNOS inhibitor had no effect on behavioral progression. The effects of nNOS genotype on behavior may be related to abnormal expression of nNOS during development, which was increased relative to controls in R6/2 mice 3 weeks of age (presymptomatic), but decreased in R6/2 mice relative to controls at 6 (around the time of symptom onset) and 11 (late-stage disease) weeks of age. Finally, protein expression of calmodulin kinase II and IV, both of which are regulators of nNOS transcription and activation, had a pattern of increased expression early in development, and decreased expression late in development, similar to that seen for nNOS. These findings indicate that nNOS activity is altered in a complex manner in HD transgenic mice and suggest that these abnormalities occur in the setting of a more global disturbance of calcium-regulated proteins.

Altered functional MRI responses in Huntington's disease.

This study examined the effects of Huntington's disease (HD) on neural activity during performance of the Porteus maze task. fMRI data were acquired from three HD patients and three controls. Reduced fMRI signal was observed in the patients relative to the controls in occipital, parietal and somato-motor cortex and in the caudate, while increased signal was found in HD in the left postcentral and right middle frontal gyri. The altered fMRI responses in HD patients may result from neural, metabolic, neurovascular coupling and/or hemodynamic differences associated with this disorder.

Biphasic developmental changes in Ca2+/calmodulin-dependent proteins in R6/2 Huntington's disease mice.

Widespread disturbances in calcium-dependent proteins are reported both in humans with advanced Huntington's disease (HD) and in symptomatic HD transgenic mice. Using a HD mouse model transgenic for exon 1 of the abnormal gene (e.g. the Bates R6/2 mouse), we found increased expression of calmodulin kinase IV and neuronal nitric oxide synthase (NOS) in 3-week-old presymptomatic HD mice striatum and cortex. Conversely, reduced expression was found at 6 weeks (early symptom onset) and 11 weeks (advanced disease) of age. The changes in protein expression may have a broad impact on the HD striatum. Calmodulin kinase IV directly regulates the activation of the transcription factors CREB (cyclic AMP response element binding protein) and CREM (cyclic AMP response element modulator) and, as well, modulates the activity of neuronal NOS. In homeostasis, nitric oxide is involved in long-term potentiation, neurotransmission, endocrine regulation and cerebral blood flow regulation, among others, while under pathological conditions nitric oxide combines with superoxide to produce the potent neurotoxin peroxynitrite. The current findings suggest that mutant HD protein may alter these processes by disturbing the regulation of calmodulin kinase IV and neuronal NOS expression across the lifespan of the HD mouse.

Reduced activity and protein expression of NOS in R6/2 HD transgenic mice: effects of L-NAME on symptom progression.

Previous work found that dietary l-arginine alters symptom progression in mice transgenic for Huntington's disease (HD), and that cerebral blood flow (CBF) is abnormal in early stage HD patients. Both of these findings potentially implicate nitric oxide (NO) and its converting enzyme, nitric oxide synthase (NOS), in HD. The current experiment found that both NOS enzymatic activity and neuronal NOS (nNOS) protein expression were reduced (P<0.05) in R6/2 HD transgenic mice compared to non-HD controls (CON). Conversely, inducible NOS (iNOS) protein expression was not significantly different between groups. The changes in nNOS were accompanied by changes in protein expression of calmodulin kinase II (CaMKII) (P<0.05) and calmodulin kinase IV (CaMKIV) (P<0.05). Protein expression of 3-nitrotyrosine (3-NT), a marker for the neurotoxin peroxynitrite, was slightly increased in non-drug treated HD and was accompanied by increased immunostaining of 3-NT in cells adhering to the vasculature and choroid plexus. Mice that received the broad-spectrum NOS inhibitor N(g)-nitro-L-arginine methyl ester hydrochloride (L-NAME) via their drinking water had reduced NOS enzyme activity. NOS activity varied as a function of L-NAME dose, was virtually eliminated in the 500-mg/l groups, and correlated (P<0.05) with the behavioral scores as revealed by regression and correlation analyses. High dose L-NAME (500 mg/l) accelerated symptom onset in HD transgenics. These results support the hypothesis that nNOS activity and NO production are abnormal in HD, this in the setting of a more global dysregulation of calcium protein expression. Taken collectively with earlier data from our laboratory demonstrating abnormal CBF findings in early-stage HD patients, these results suggest that abnormalities in NOS function may significantly contribute to the neurodegeneration found in HD.

Nitric oxide and nitric oxide synthase in Huntington's disease.

Nitric oxide (NO) is a biologically active inorganic molecule produced when the semiessential amino acid l-arginine is converted to l-citrulline and NO via the enzyme nitric oxide synthase (NOS). NO is known to be involved in the regulation of many physiological processes, such as control of blood flow, platelet adhesion, endocrine function, neurotransmission, neuromodulation, and inflammation, to name only a few. During neuropathological conditions, the production of NO can be either protective or toxic, dependent on the stage of the disease, the isoforms of NOS involved, and the initial pathological event. This paper reviews the properties of NO and NOS and the pathophysiology of Huntington's disease (HD). It discusses ways in which NO and NOS may interact with the protein product of HD and reviews data implicating NOS in the neuropathology of HD. This is followed by a synthesis of current information regarding how NO/NOS may contribute to HD-related pathology and identification of areas for potential future research.

Dietary arginine alters time of symptom onset in Huntington's disease transgenic mice.

Recent neuroimaging studies reported complex changes in cerebral blood flow (CBF) in early-staged Huntington's disease (HD) patients. Deckel and co-workers [Deckel and Duffy, Brain Res. (in press); Deckel and Cohen, Prog. Neuro-Psychopharmacol. Biol. Psychiatry 24 (2000) 193; Deckel et al., Neurology 51 (1998) 1576; Deckel et al., J. Nucl. Med. 41 (2000) 773] suggested that these findings might be accounted for, in part, by alterations in cerebral nitric oxide (NO) and its byproduct, peroxynitrite. The current experiment tested this hypothesis by altering NO levels via manipulations of dietary L-arginine (ARG), the dietary precursor of NO, in mice transgenic for HD. Seventy-one mice were assigned at 12 weeks of age to one of three isocaloric diets that varied in their content of ARG. These diets included: (a) 0% ARG, (b) 1.2% ARG (i.e. typical mouse chow), or (c) 5% ARG. The 5% ARG diets in HD mice accelerated the time of onset of body weight loss (P<0.05) and motor impairments (P<0.05), and increased resting CBF in HD relative to control (P<0.05). Conversely, the 0% ARG diet demonstrated no loss of body weight and had no changes in CBF relative to controls. However, the 0% ARG HD group continued to show significant deficits on motor testing (P<0. 05). The 1.2% ARG HD group showed reduced body weight loss, better motor functioning, and fewer changes in CBF compared to the 5% ARG HD group. Immunocytochemistry analysis found greater deposition of nitrotyrosine in the cortex, and vasculature, of HD+ mice, 5% and 1. 2%>0% arginine diets. When collapsed across all conditions, CBF inversely correlated (P<0.05) both with the body weight and motor changes suggesting that changes in CBF are associated with behavioral decline in HD mice. Collectively, these findings indicate that dietary consumption of the NO precursor ARG has a measurable, but complex, effect on symptom progression in HD transgenic mice, and implicates NO in the pathophysiology of HD.

Vasomotor hyporeactivity in the anterior cerebral artery during motor activation in Huntington's disease patients.

This experiment used functional transcranial Doppler ultrasonography to demonstrate that blood flow velocity in the anterior cerebral artery is hyporeactive in Huntington's disease (HD) patients during maze testing (P<0.05). These vascular changes are due to tracing, as opposed to problem solving, components of the maze task. Using logistic regression, the reactivity of the ACA during activation is able to categorize patients as being either gene negative, or positive, for HD (P=0.0007). The possible role that nitric oxide/peroxynitrite may play in this phenomena is discussed.

Increased CBF velocity during word fluency in Huntington's disease patients.

1. This study examined the effects of word fluency and reading on cerebral blood flow in Huntington's disease (HD) patients. 2. Changes in cerebral flow velocity in the anterior (ACA) and middle (MCA) cerebral arteries were measured with functional transcranial Doppler ultrasonography (fTCD) in 13 normal controls and 9 gene positive HD patients. To control for motor effects of word fluency, two "control" conditions, including silent word fluency and a reading test, were also administered to all subjects. 3. Cerebral blood flow velocity was increased during the out loud word fluency test in the ACA, but not MCA, in the HD group compared to controls. This increase was due to motor components of the test, as during silent word fluency the HD group had a decrease in cerebral blood flow relative to controls. Significant correlations between blood flow in the ACA and word fluency test scores were found. Cerebral blood flow velocity during testing also was able to predict group assignment (i.e., control vs. mild HD vs. moderate HD). 4. These findings add to a growing body of literature suggesting that CBF velocity in HD is abnormal during cognitive and motor tasks. Although previous work reported that CBF velocity in HD is decreased during hand use on a maze test, the current experiment finds that speech production increases cerebral blood flow velocity in HD patients. Collectively, these results point to a fundamental disturbance in the regulation of CBF in HD. Mechanisms that could account for these findings, including the potential involvement of nitric oxide, are discussed.

Altered patterns of regional cerebral blood flow in patients with Huntington's disease: a SPECT study during rest and cognitive or motor activation.

UNLABELLED: Previous research using functional transcranial Doppler sonography showed that blood flow velocity in the anterior cerebral artery is significantly less in patients with Huntington's disease (HD) than in healthy volunteers while they are completing mazes. The current research used SPECT to study regional cerebral blood flow (rCBF) in patients with HD during rest and maze testing. METHODS: Seven patients with HD and 9 healthy volunteers were injected twice with 0.96-1.15 GBq 99mTc-labeled hexamethylpropylene amine oxime. During the 10 min after injection, subjects either solved mazes or rested with their eyes open while looking at a modified maze. After SPECT, count density was obtained from 11 brain regions and corrected for decay and injected dose. Two types of data generated from this experiment, including absolute regional counts per pixel in the regions of interest and count density computed as a percentage of activity in the lateral cerebellum, were compared between groups. RESULTS: During rest, the absolute regional count density was greater in the HD brains than in the healthy brains (P < 0.001). Count density was typically between 8% and 13% higher in the HD group than in the healthy group. The single exception was the caudate density, for which the 2 groups had similar values. No significant differences in absolute regional count density were observed between groups during maze testing. When rCBF was calculated as a percentage of cerebellar rCBF, analysis of covariance found decreases in HD caudate density (P < 0.001) and orbital frontal cortex density (P < 0.005) during maze testing. Changes in rCBF in the caudate nucleus predicted gene status (P = 0.0007) and correlated with time to complete the mazes (P < 0.05). CONCLUSION: Patients with HD showed an increase in resting rCBF for all brain regions measured except the caudate nucleus. When rCBF was calculated as a percentage of cerebellar blood flow, rCBF in the striatum and orbital cortex in patients with HD was less during maze testing than during rest. Although the cause of these rCBF changes in HD patients is unclear, nitric oxide synthase, a regulator of vasomotor activity, may be involved.

Tests of executive functioning predict scores on the MacAndrew Alcoholism Scale.

1. Previous work reported that tests of executive functioning (EF) predict the risk of alcoholism in subject populations selected for a "high density" of a family history of alcoholism and/or the presence of sociopathic traits. The current experiment examined the ability of EF tests to predict the risk of alcoholism, as measured by the MacAndrew Alcoholism Scale (MAC), in outpatient subjects referred to a general neuropsychological testing service. 2. Sixty-eight male and female subjects referred for neuropsychological testing were assessed for their past drinking histories and administered the Wisconsin Card Sorting Test, the Wechsler Adult Intelligence Scale-Revised, the Trails (Part B) Test, and the MAC. Principal Components analysis (PCA) reduced the number of EF tests to two measures, including one that loaded on the WCST, and one that loaded on the Similarities, Picture Arrangement, and Trails tests. Multiple hierarchical regression first removed the variance from demographic variables, alcohol consumption, and verbal (i.e., Vocabulary) and non-verbal (i.e., Block Design) IQ, and then entered the executive functioning factors into the prediction of the MAC. 3. Seventy-six percent of the subjects were classified as either light, infrequent, or non-drinkers on the Quantity-Frequency-Variability scale. The factor derived from the WCST on PCA significantly added to the prediction of risk on the MAC (p = .0063), as did scores on Block Design (p = .033). Relatively more impaired scores on the WCST factor and Block Design were predictive of higher scores on the MAC. The other factors were not associated with MAC scores. 4. These results support the hypothesis that decrements in EF are associated with risk factors for alcoholism, even in populations where the density of alcoholic behaviors are not unusually high. When taken in conjunction with other findings, these results implicate EF test scores, and prefrontal brain functioning, in the neurobiology of the risk for alcoholism.

Cerebral blood flow velocity decreases during cognitive stimulation in Huntington's disease.

OBJECTIVE: To examine whether activation of cerebral blood flow velocity during cognitive stimulation, as measured by transcranial Doppler ultrasonography (TCD), is altered in patients with Huntington's Disease (HD). BACKGROUND: Previous research suggests that resting cerebral blood flow in symptomatic and asymptomatic HD patients is reduced from expected premorbid levels. The effects of cognitive activation on this relative hypoperfusion in HD has not been studied extensively. METHODS: We measured TCD flow velocity during rest and cognitive testing with the Porteus Maze Test and the Trails Test in 12 normal control subjects and 10 gene-positive HD patients. Percent change (i.e., flow during testing/resting) of flow velocity in the anterior and middle cerebral arteries were compared between groups. Correlations among percent flow velocity change, a disability rating scale, and cognitive test scores were calculated. RESULTS: In control subjects, anterior cerebral artery flow velocity and middle cerebral artery velocity increased during cognitive testing (p=0.001). HD patients showed a smaller blood flow velocity increase in the anterior cerebral arteries during the Porteus Maze Test (p < 0.001) and the Trails Test, Part B (p < 0.001). In the left anterior cerebral artery, flow velocity fell an average of 4.2% below resting baseline levels during the Porteus Maze Test, and 1.2% below resting levels during the Trails Test. The magnitude of the cerebral blood flow velocity changes in the anterior cerebral artery correlated with a number of cognitive test scores and with a rating scale of functional disability. In addition, logistic regression was able to discriminate the HD patients from the control group based on blood flow velocity changes (p=0.0025). When HD patients were divided into more (i.e., HD with chorea; n=4) and less impaired (i.e., without chorea; n=6) groups, both showed significant decreases in left anterior cerebral artery flow velocity during visual spatial executive cognition testing compared with control subjects. CONCLUSIONS: These results suggest that activation of visual spatial executive functions cause decreased flow velocity in the anterior cerebral artery, but not the middle cerebral artery, in HD patients. These changes are related to test performance and functional capabilities.

Effects of serotonergic drugs on lateralized aggression and aggressive displays in Anolis carolinensis.

Previous work demonstrated that the brains of many reptiles, including the American chameleon Anolis carolinensis (A. carolinensis), are functionally 'split'. Because the left eye in this species projects predominantly to the right hemisphere, and vice versa, inferences about lateralized brain functioning can be made in A. carolinensis by observation of eye use during behavioral encounters. Using this model, past work suggested that territorial aggression in Anolis is under the preferential control of the right hemisphere, and that acute stress or chronic alcohol exposure selectively reduces right hemisphere mediated territorial aggression. In addition, drugs which increase serotonin (5-HT) in the synaptic cleft inhibit aggressive responding in anoles in both hemispheres. The current experiment examined whether or not the administration of the serotonin agonists 8-hydroxy-2-(di-n-propylamine) tetralin (8-OHDPAT), quipazine, or meta-chlorophenylbiguanide (mCPBG) alter territorial aggression in Anolis. Nine adult socially isolated male A. carolinensis underwent a series of behavioral trials during which an antagonistic male was introduced into the cage. Once stable responding was initiated, all subjects were injected in a semi-randomized crossover manner with the following agents, (1) lactated Ringer's, (2) the 5-HT2 agonist quipazine (1.5 mg/kg and 3.0 mg/kg), (3) the 5-HT1 agonist 8-OHDPAT (83 mg/kg), and (4) the 5-HT3 agonist mCPBG (3.0 mg/kg and 9 mg/kg). Twenty minutes post injections, the male intruder was reintroduced into the subject's cage. Several behaviors were recorded, including: (1) the time to the first aggressive response, (2) the number of aggressive episodes mediated by the left eye or right eye, and (3) changes in skin color and posture. Aggressive responding was virtually eliminated in all subjects injected with 8-OHDPAT. On the other hand, one-way ANOVA found that both the 9 mg/kg dose of mCPBG (P=0.007), and the 3.0 mg/kg dose of quipazine (P=0.035), selectively decreased territorial aggression mediated by the left eye/right hemisphere compared to lactated Ringer's controls, but had no effect on aggression mediated by the right eye/left hemisphere. Although 8-OHDPAT inhibited aggression, injected subjects developed phenotypic displays of aggressive coloring/posturing, such as blackening of the eye spot and a raising of the neck crest. These results suggest that aggressive action can be differentiated from phenotypic displays that accompany aggression by a 5-HT1 agonist. They also indicate that there is an asymmetrical effect of 5-HT2/5-HT3 serotonin agonists on hemispheric mediation of aggression in this species.

Lateralized effects of ethanol on aggression and serotonergic systems in Anolis carolinensis.

The lateralized effects of ethanol (ETOH) upon behavior and monoamine biochemistry in the lizard, Anolis carolinensis, were examined. Eight adult male anoles consumed solutions of 19% ethanol (ETOH) twice daily over the course of 18 days, while controls consumed water. ETOH decreased the use of the left eye/right hemisphere, but not the right eye/left hemisphere, during territorial aggression (p<0.05). During crossover (i.e., ETOH to water and vice versa) this effect was reversible and replicable. Biochemically, an asymmetry was observed in 5-HT levels in the raphe both in ETOH and controls. ETOH increased levels of serotonin (5-HT; p<0.05), and 5-HIAA/5-HT ratios (p<0.05) in the raphe; serotonin levels in several brain regions correlated with aggressive responses. These results suggest that ETOH boosts 5-HT levels in animals subchronically exposed to ETOH. They further suggest that asymmetry in endogenous 5-HT systems may account for the asymmetrical regulation of aggression generally, and may explain the behavioral effects of ETOH upon lateralized aggression.

Hemispheric control of territorial aggression in Anolis carolinensis: effects of mild stress.

Previous work has demonstrated that the brain of Anolis carolinensis is functionally split, in that the left and right eyes project predominantly to the contralateral hemisphere, and as there are minimal connections between the left and right hemispheres. Using this model, the current experiment examined the effect of mild acute stress on hemispheric regulation of territorial aggression. Thirteen adult male Anolis were paired with an antagonistic males, and eye use and behavioral responses were repeatedly measured during 3 minute behavioral trials. Trials were conducted either after exposure to mild stress, produced by handling the subject, or without stress, and they were run either in the subject's home cage or in a cage foreign to the subject. Left eye preference for aggressive movements was found during the trials run in the non-stressed conditions (p < 0.05). Conversely, stressed subjects showed a reduction in left eye/right hemisphere mediated aggressive movements relative to the non-stressed subjects but no changes in right eye/left hemisphere aggression. This effect was independent of whether or not the subject was in its home or a foreign cage. No laterality in aggressive responding was found when the subjects were placed in separate cages with visual contact. These findings suggest that territorial aggression in Anolis is preferentially initiated and processed by the left eye/right hemisphere but is subject to right-hemispheric inhibition following exposure to acute mild stress.

Effects of 5,7-dihydroxytryptamine lesions of the prefrontal cortex on consumption of sucrose-ethanol solutions: relationship to prefrontal monoamines.

Thirty adult male Wistar rats received 8 micrograms bilaterally of 5,7-dihydroxytryptamine into the medial prefrontal cortex (mPFC). Rats were then trained, via a sucrose, fading paradigm, to consume increasing concentrations of alcohol. After death, dopamine (DA), norepinephrine (NE), serotonin (5-HT), and their metabolites were measured in the mPFC, nucleus accumbens (NA), and raphe nucleus. The lesioned group demonstrated a reduction in 5-hydroxyindoleacetic acid (5-HIAA), DA, and NE in the mPFC (p < 0.05), and a trend toward reduction of 5-HT in the NA. In comparison with controls, lesioned animals consumed less of all solutions containing sucrose and alcohol. On regression analyses, monoamines in the mPFC (i.e., 5-HIAA, dihydrophenylacetic acid and NE) predicted consumption of the 5% ethanol solution (p = 0.009), 10% ethanol solution (p = 0.0006), and the 5% sucrose solutions (p = 0.0006), but not the 20% sucrose solutions. In each case, monoamine levels were positively correlated with consumption. No relationships were seen between monoamine levels in the NA and raphe, and in consummatory behavior.

Effects of alcohol consumption on lateralized aggression in Anolis carolinensis.

Previous work has suggested that the lizard Anolis carolinensis, like many other reptiles, has a functionally split brain. Specifically, the left eye of this species projects primarily to the right hemisphere (and vice versa), there is no corpus callosum, and the physical placement of the eyes restricts their field(s) of vision to one region of hemispace. The current experiment used this preparation to examine the effect of alcohol administration and withdrawal on lateralized brain functioning during territorial aggression. Thirteen adult males were divided into control (CON) or alcohol (ETOH) groups. Baseline territorial aggression was assessed, following which ETOH subjects were then given twice daily solutions of 19% alcohol. After 19 days of ETOH consumption, territorial aggression was again assessed. Eye use during aggressive encounters was measured either following short periods (24 h) of alcohol withdrawal, or 90 m following alcohol consumption. Control subjects were found to have a predominance of left eye/right hemisphere-mediated aggressive responses, as has previously been reported. Alcohol withdrawn subjects were found to have a suppression of left eye/right hemisphere-mediated aggression. This reached statistical significance on several measures of aggression, including the number of dewlaps and headbob (P < 0.001) and the total number of aggressive responses (P = 0.001). Consumption of ETOH reversed this pattern and reinstated the normal pattern of left eye/right hemisphere dominance during aggression. Conversely, right eye/left hemisphere mediation of aggression was found to be increased, or not affected, during alcohol withdrawal, and to show no differences from CON following ETOH administration. Extrapolating from other recent findings in this species, these results suggest that the stress caused by ETOH withdrawal on the CNS may differentially effect the right hemisphere of the brain while having little effect on the left.

Antisocial personality disorder, childhood delinquency, and frontal brain functioning: EEG and neuropsychological findings.

This experiment examined the ability of EEG activity and neuropsychological testing to predict both antisocial personality disorder (ASP) and retrospective self-ratings of early childhood problem behaviors (CPB). Regression analyses found that increased frontal left-hemisphere EEG activation was associated with a decreased likelihood of the diagnosis of ASP or CPB. An association was also found between several motor tests of the Luria-Nebraska and Porteus Maze Test scores and CPB/ASP. The current findings suggest that ASP and CPB are associated with variations in frontal lobe functioning. They further suggest that disturbances in prefrontal functioning may be a common biological ground that links ASP, substance abuse, and biological mechanisms of reinforcement.

Behavioral and cognitive measurements predict scores on the MAST: a 3-year prospective study.

This experiment examined the ability of neuropsychological and behavioral tests of anterior brain functioning to predict changes in alcohol-related behaviors 3 years after the initial evaluation. One hundred four young adult subjects, sixty-six of whom had a positive family history of alcoholism, filled out the Michigan Alcoholism Screening Test (MAST), self-rated the frequency with which they consumed alcohol, and completed a retrospective test of childhood hyperactivity/impulsivity/conduct problems (Wender Behavioral Checklist). Eighty-three of the subjects also completed a neuropsychological battery consisting of cognitive tests of executive functioning (i.e., Trails, Categories Test, Similarities) and motor tests (i.e., TPT, hand dynamometer, finger tapping). Subjects returned to complete the alcohol-related measures during a second testing session 3 years later. Changes in MAST scores, and in alcohol consumption, between the first and second test session were computed, and median splits classified subjects into "high" versus "low" change groups. After factor analysis of the cognitive and behavioral data, hierarchical logistic regression equations assessed the ability of the cognitive and behavioral variables, as well as the presence of a positive family history of alcoholism, to predict future changes in the alcohol outcome measures. Scores from the WENDER behavioral checklist predicted future changes in the MAST scores (p = 0.0026), with more impaired scores associated with higher MAST scores. Tests of executive functioning, in subjects with a positive family history of alcoholism, predicted alcohol consumption (p = 0.033). None of the other predictor variables showed any relationship to the alcohol-related outcome measures.

Behavioral changes in Anolis carolinensis following injection with fluoxetine.

Eight adult male lizards of the genus and species Anolis carolinensis were used in this experiment. In order to induce aggressive responding, animals were caged separately and daily underwent pairing with another male, during which aggressive responses and changes in skin color were measured. After obtaining a baseline measure of aggressive responding, animals were injected either with fluoxetine or vehicle-controls in a cross-over design. Subjects were then exposed to five more days of (non drug) pairing with the intruder male, after which they underwent a second trial with fluoxetine/vehicle. Finally, two post-drug paired-trials were obtained. Fluoxetine injection significantly reduced the aggressive responding in the males while causing the postorbital eyespot to significantly darken. Subjects also showed increased aggressivity and skin-color reactivity subsequent to the two drug trials, although it is unclear if the fluoxetine, or non-specific factors of the injection paradigm, accounted for these changes. These results suggest that serotonergic CNS systems tonically regulate aggression in Anolis carolinesis, similar to that seen in many other species. They further suggest that eyespot-darkening and aggressive responding can be pharmacologically dissociated, implicating serotonin in the regulation of this phenomenon.