Neurokinin-1 receptor deletion modulates behavioural and neurochemical alterations in an animal model of depression.

The substance P/NK1 receptor system plays an important role in the regulation of stress and emotional responding and as such had been implicated in the pathophysiology of anxiety and depression. The present study investigated whether alterations in the substance P/NK1 receptor system in brain areas which regulate emotional responding accompany the depressive behavioural phenotype observed in the olfactory bulbectomised (OB) mouse. The effect of NK1 receptor deletion on behavioural responding and monoamine levels in discrete brain regions of the OB model, were also examined. Substance P levels in the frontal cortex and NK1 receptor expression in the amygdala and hippocampus were enhanced following olfactory bulbectomy. Although NK1 receptor knockout (NK1-/-) mice did not exhibit altered behavioural responding in the open field test, noradrenaline levels were enhanced in the frontal cortex, amygdala and hippocampus, as were serotonin levels in the frontal cortex. Locomotor activity and exploratory behaviour were enhanced in wild type OB mice, indicative of a depressive-like phenotype, an effect attenuated in NK1-/- mice. Bulbectomy induced a decrease in noradrenaline and 5-HIAA in the frontal cortex and an increase in serotonin in the amygdala, effects attenuated in OB NK1-/- mice. The present studies indicate that alterations in substance P/NK1 receptor system underlie, at least in part, the behavioural and monoaminergic changes in this animal model of depression.

The experimental induction of leukoencephalomyelopathy in cats.

Leukoencephalomyelopathy of undetermined etiology has been described in specific pathogen-free cats. A study was established to assess if the long-term feeding of a gamma-irradiated diet could induce this disease. Cats fed exclusively on diet irradiated at 25.7-38.1 kGy ("typical" dose) and 38.1-53.6 kGy (high-end dose), respectively, developed typical lesions with attendant, progressively severe ataxia between study days 140 and 174. The onset of ataxia at day 140 and the number of animals affected at this time were similar in animals fed each ration. A maximum ataxia "score" was first reached by an animal on the high-end dose diet on day 167 and by 2 cats fed the "typical-end" dose diet 21 days later. Ataxic cats and 1 animal euthanized on day 93 prior to the onset of ataxia exhibited varying degrees of Wallerian degeneration in the spinal cord and brain, similar to the spontaneous disease. The elevated total antioxidant status of spinal cord segments and hepatic superoxide dismutase concentration of cats fed typical and high-end treated diets suggested free-radical involvement in the pathogenesis. The significantly elevated peroxide concentrations of the irradiated diets (1,040% and 6,440% of untreated values) may have resulted in increased oxidative insult, a factor possibly exacerbated by the treated diets' reduced vitamin A content. This study has reproduced leukoencephalomyelopathy in cats similar to spontaneous outbreaks by feeding a gamma-irradiated dry diet with elevated peroxide and reduced vitamin A concentrations.

Effect of level and duration of dietary n-3 polyunsaturated fatty acid supplementation on the transcriptional regulation of Delta9-desaturase in muscle of beef cattle.

The objective of this study was to examine the effect of level and duration of feeding of an n-3 PUFA-enriched fish oil (FO) supplement in combination with soybean oil (SO) on the transcriptional regulation of Delta(9)-desaturase gene expression in bovine muscle. Beef bulls (n = 40) were assigned to 1 of 4 iso-lipid and isonitrogenous concentrate diets fed for ad libitum intake for a 100-d finishing period. Concentrates were supplemented with one of the following: 1) 6% SO (CON); 2) 6% SO + 1% FO (FO1); 3) 6% SO + 2% FO (FO2); or 4) 8% palmitic acid for the first 50 d and 6% SO + 2% FO for the second 50 d [FO2(50)]. Samples of LM were harvested and concentrations of fatty acids were measured. Total RNA was isolated and the gene expression of Delta(9)-desaturase was determined. The mRNA expression of putative regulators of Delta(9)-desaturase gene expression, sterol regulatory element binding protein-1c (SREBP-1c) and peroxisome proliferator activated receptor-alpha (PPAR-alpha), were also measured in the CON and FO2 groups. Expression of mRNA for Delta(9)-desaturase was decreased (P < 0.05) 2.6-, 4.4-, and 4.9-fold in FO1, FO2(50), and FO2 compared with CON, respectively. Expression of Delta(9)-desaturase mRNA tended to be reduced (P = 0.09) by increasing FO from 1 to 2%, but was not affected by duration of supplementation (P > 0.24). Expression of mRNA for SREBP-1c was decreased 2-fold in FO2 compared with CON (P < 0.05), whereas expression of PPAR-alpha was not affected (P > 0.30). There was a positive relationship between Delta(9)-desaturase and SREBP-1c gene expression (P < 0.01), but the expression of both genes was negatively related to tissue concentrations of n-3 PUFA (P < 0.05) and positively related to concentration of n-6 PUFA (P < 0.01). Simultaneous enhancement of tissue concentrations of CLA and n-3 PUFA concentrations in bovine muscle may be hindered by negative interactions between n-3 PUFA and Delta(9)-desaturase gene expression, possibly mediated through reduced expression of SREBP-1c.

Effect of traumatic subarachnoid haemorrhage on neuropsychological profiles and vocational outcome following moderate or severe traumatic brain injury.

OBJECTIVE: Traumatic subarachnoid haemorrhage (tSAH) frequently occurs in moderate or severe traumatic brain injury (TBI) and is related to worse outcome at time of discharge from the acute hospitalization. The current study compared neuropsychological impairment and vocational outcome at 1-year post-injury in patients with and without tSAH. DESIGN: Acute injury, neuroradiological, neuropsychological and vocational data were collected for 100 patients admitted for neurorehabilitation following TBI. RESULTS: Patients with tSAH had significantly worse vocational outcome than patients without tSAH. On neuropsychological measures, patients with tSAH generally performed worse than patients without tSAH across most neurocognitive domains. However, differences in neuropsychological test performance between patients with and without tSAH reached statistical significance on measures of visuospatial processing, verbal reasoning and mood only. CONCLUSION: The presence of tSAH appears to be associated with worse vocational outcome in survivors of moderate or severe TBI. As such, the presence of tSAH appears to have predictive value with respect to outcome following TBI.

Acuity, ophthalmoscopy, and visually evoked potentials in the prediction of visual outcome in infants with bilateral optic nerve hypoplasia.

PURPOSE: To determine whether Teller Acuity Cards, transient visually evoked potentials (VEPs), and optic disc size estimated from ophthalmoscopy were predictive of acuity outcome in infants and young children with bilateral optic nerve hypoplasia (ONH). METHODS: Twenty-eight infants (mean age, 7 months) with bilateral ONH underwent clinical assessment, including ophthalmoscopy. All but a few of these patients underwent neuroimaging studies, analysis of transient VEPs to multiple stimuli, and repeated acuity assessment. Acuity outcome was assessed, on average, 28 months later. RESULTS: Acuity outcome was significantly correlated with the estimated optic disc diameter, initial acuity, and VEP signal-to-noise ratios (r = 0.80, 0.71, 0.69, respectively; all P <.001). Multiple regression analysis showed that the initial acuity and estimated optic disc diameter accounted for 73% of the variation in acuity outcome. VEPs to white-black gratings segregated infants by 6 months of age, whose acuity outcome was better or worse than 5.6 cycles/degree (20/100). CONCLUSIONS: Acuity outcome was predicted in infants with bilateral ONH with a linear equation using initial acuity and estimated optic disc diameter. Additionally, analysis of VEPs may segregate infants with a good visual outcome from those with a poor visual outcome. Longer follow-up will be necessary to determine final Snellen acuity.

Serotonergic mediation of the antidepressant-like effects of nitric oxide synthase inhibitors.

Recent studies indicate that nitric oxide (NO) synthase inhibitors have antidepressant-like potential in various animal models. In the present study the behavioural activity of the NO synthase inhibitors, N(G)-nitro-L-arginine (L-NA) and 7-nitroindazole (7-NI), were assessed in a modified rat forced swimming test (FST). Both L-NA and 7-NI, dose dependently reduced immobility and increased swimming behaviour in the rat FST. This behavioural profile parallels the one previously shown with selective serotonin re-uptake inhibitors and serotonergic agonists. Thus, we examined the role of serotonin mediating the behavioural effects of L-NA and 7-NI in the rat FST. Depletion of endogenous serotonin using para-chlorophenylalanine (pCPA; 3 x 150 mg/kg, i.p.) completely blocked L-NA (20 mg/kg, i.p.) and 7-NI (20 mg/kg, i.p.)-induced reductions in immobility and increases in swimming behaviour during the FST. In conclusion these observations suggest that NO synthase inhibitors elicit their antidepressant-like activity in the modified swimming test through a serotonin dependent mechanism.

Effects of acute and chronic administration of selective monoamine re-uptake inhibitors in the rat forced swim test.

The rat forced swim test (FST) is a model that is used extensively as a screening test for antidepressant activity. It has previously been reported that thorough analysis of behaviour in this model reveals two distinct types of active response - climbing and swimming - and that these are separately evoked by re-uptake inhibitors selective for noradrenaline (NA) and serotonin (5-HT), respectively. In the present study, utilising re-uptake inhibitors selective for NA, talsupram, and 5-HT, 5-chloro-1-(3-dimethylaminopropyl)-1-(4-fluorophenyl)- phthalan (Lu 10-134-C), we examined if this scoring technique could detect the antidepressant potential of a selective serotonin re-uptake inhibitor (SSRI), and whether re-uptake inhibitors selective for distinct monoamine systems induce exclusive behavioural responses. We also analysed if chronic antidepressant administration for three weeks was more effective than acute treatment. We found Lu 10-134-C (40 mg/kg; PO) to be behaviourally active in this paradigm. Although treatment with talsupram (40 mg/kg; PO) resulted solely in climbing behaviour, Lu 10-134-C induced both climbing and swimming behaviour. However, chronic pre-treatment with either re-uptake inhibitor (20 mg/kg; twice daily; PO) failed to augment the response observed with acute treatment. Similarly, chronic administration of either compound was without effect on the basal, or stress-induced, serum corticosterone concentrations or anterior pituitary (AP) preproopiomelanocorticotropin (POMC) mRNA expression. These results suggest that selective monoamine re-uptake inhibition produces distinct, but not necessarily exclusive, behavioural responses in the forced swim test.

The effect of lofepramine and other related agents on the motility of Tetrahymena pyriformis.

Tricyclic antidepressants (TCAs) were introduced almost 50 years ago. Whilst there is no doubt that TCAs are effective in treating depression, they are also more cardiotoxic when taken in overdose than other antidepressant groups. Lofepramine is a more recently introduced modified TCA, which in animals and man has low toxicity when compared to older TCAs. Paradoxically, lofepramine is extensively metabolised to desipramine, which has considerable toxicity, both experimentally and in overdose. The toxicity of such compounds is attributed, in part, to a membrane stabilising effect (MSA) on cell membranes. This MSA causes gross effects to the cell structure and in turn, normal cell activity. The aim of this study was to compare the MSA of lofepramine with that of desipramine and amitriptyline in order to see if this might help to explain the low toxicity of lofepramine. The local anaesthetic agent lignocaine was also studied for comparison. Each compound was enclosed in a beta-cyclodextrin to increase its solubility in aqueous medium. The extent of MSA was determined as a measure of the effect on the swimming speed of the protozoan Tetrahymena pyriformis using a video image analysis system. The IC50s for the various drugs were then correlated with their respective octanol-water partition coefficient values (Pow). Amitriptyline had an IC50 of 1.26+/-0.29 mM, desipramine 75.99+/-14.40 mM, while lofepramine had an IC50 of 357.40+/-25.00 mM. Lignocaine had an IC50 of 85.73+/-18.30 mM. There was also a significant correlation between the IC50 values and the Pow values.

The minimally conscious state: definition and diagnostic criteria.

OBJECTIVE: To establish consensus recommendations among health care specialties for defining and establishing diagnostic criteria for the minimally conscious state (MCS). BACKGROUND: There is a subgroup of patients with severe alteration in consciousness who do not meet diagnostic criteria for coma or the vegetative state (VS). These patients demonstrate inconsistent but discernible evidence of consciousness. It is important to distinguish patients in MCS from those in coma and VS because preliminary findings suggest that there are meaningful differences in outcome. METHODS: An evidence-based literature review of disorders of consciousness was completed to define MCS, develop diagnostic criteria for entry into MCS, and identify markers for emergence to higher levels of cognitive function. RESULTS: There were insufficient data to establish evidence-based guidelines for diagnosis, prognosis, and management of MCS. Therefore, a consensus-based case definition with behaviorally referenced diagnostic criteria was formulated to facilitate future empirical investigation. CONCLUSIONS: MCS is characterized by inconsistent but clearly discernible behavioral evidence of consciousness and can be distinguished from coma and VS by documenting the presence of specific behavioral features not found in either of these conditions. Patients may evolve to MCS from coma or VS after acute brain injury. MCS may also result from degenerative or congenital nervous system disorders. This condition is often transient but may also exist as a permanent outcome. Defining MCS should promote further research on its epidemiology, neuropathology, natural history, and management.

The infant who is visually unresponsive on a cortical basis.

OBJECTIVE: To further explain cortical abnormalities in infants without visually guided behaviors with anatomically normal eyes by using a battery of objective visual tests and neuroimaging studies. DESIGN: A cohort study. PARTICIPANTS: Thirty-one infants with clear ocular media and normal fundi, who were visually unresponsive by clinical examination, and 31 control subjects. METHODS: Full clinical examinations including Teller Acuity Cards (TAC) and developmental assessment. Infants with reduced acuities and/or developmental delay underwent pattern visual evoked potential (VEP) testing and brain neuroimaging studies. Eye movement recordings were done in individual infants. MAIN OUTCOME MEASURES: Visual acuity, VEPs, eye movement recordings. RESULTS: Infants were separated into two groups on the basis of being developmentally normal (DN) or developmentally delayed (DD). Fourteen DN infants had normal acuities for age and three of three infants had normal VEPs. On the basis of having normal visual function, these infants were considered to have visual inattention (VI). Sixteen DD infants had acuities ranging from normal to no visual orienting to the low vision TAC. All 16 DD infants had abnormal VEPs and abnormal neuroimaging studies (brain computed tomography, magnetic resonance imaging, or both) or microcephaly. On the basis of having structural and functional abnormalities of the brain, these infants were diagnosed as having cortical visual impairment (CVI). One additional infant with DD failed to orient to TACs but had a normal VEP and normal magnetic resonance imaging. In this infant and two infants with CVI the inability to "fix and follow" was attributed completely or partially to an oculomotor apraxia (OMA), which was confirmed by eye movement recordings. CONCLUSIONS: The infant who is visually unresponsive on a cortical basis has either VI or CVI. Infants with OMA can also seem to be visually unresponsive. These disorders can be delineated in infancy on the basis of developmental status and a unique set of responses to visual acuity, VEPs, and oculomotor testing.

Methylenedioxymethamphetamine (MDMA; 'Ecstasy') suppresses antigen specific IgG2a and IFN-gamma production.

Methylenedioxymethiamphetamine (MDMA; "Ecstasy") is a widely abused amphetamine derivative. In the present study, we examined the effect of acute MDMA administration on an antigen specific immune response. Responsiveness to an in vivo challenge with the soluble protein antigen keyhole limpet haemocyanin (KLH) was examined in rats following MDMA administration (2.5, 5 or 10 mg/kg; i.p.). KLH-specific serum IgM concentrations were measured 7 days following challenge, and serum IgG concentrations were measured 14 days following the KLH challenge. In addition, antigen-specific IFN-gamma and IL-6 production was measured in KLH-stimulated splenocytes. MDMA did not alter the KLH-specific IgM response. In contrast, MDMA (5 and 10 mg/kg) provoked a significant suppression of KLH-specific IgG production. Thus, MDMA administration did not alter the initial generation of the antibody response but rather inhibited antibody class switching from IgM to IgG. Two pathways for the genetic switch from IgM to IgG production were investigated. One pathway requires the Th(1) type cytokine IFN-gamma to stimulate IgM-secreting cells to switch to IgG(2a)-secreting cells. Another pathway requires the Th(2) type cytokines IL-4 and IL-6 to stimulate IgM-secreting cells to switch to IgG(1)-secreting cells. IgG(1) and IgG(2a) levels were measured to determine if these two pathways were differentially affected. The results indicate that only IgG(2a) levels were decreased following MDMA administration. Furthermore, this decrease in IgG(2a) was accompanied by decreased KLH-specific IFN-gamma production 14 days post KLH administration. In conclusion, these data indicate that MDMA alters the ability to switch from IgM to IgG(2a) production, possibly by reducing IFN-gamma. Potential health consequences for MDMA users are discussed.

The virtual retinal display as a low-vision computer interface: a pilot study.

This pilot study examined the performance of an alternative computer visual interface, the Virtual Retinal Display (VRD), for low-vision use. The VRD scans laser light directly onto the retina, creating a virtual image. Since visually impaired individuals can have difficulty using computer displays, a matched comparison study was done between the VRD and the standard cathode ray tube (CRT) monitor. Reading speed and acuity tests were collected from 13 low-vision volunteers selected to represent the broad range of partially sighted individuals actively involved in the work force. Forty-six percent of subjects had highest visual acuity while viewing the VRD; 30% of subjects had highest acuity viewing the CRT; and 24% of subjects had equal acuity across the two displays. Although mean reading speed across all 13 subjects indicated no significant difference between displays, individual subjects with predominantly optical causes of low vision exhibited clinically important increases in reading speed versus the CRT. However, most subjects with predominantly retinal damage showed a slight disadvantage using the VRD. We give theoretical explanation to the bifurcated results and conclude that for a subset of low-vision users, the VRD technology is very promising as a basis for future low-vision aids.

Beta-amyloid(1-42)-induced cholinergic lesions in rat nucleus basalis bidirectionally modulate serotonergic innervation of the basal forebrain and cerebral cortex.

Ample experimental evidence suggests that beta-amyloid (A beta), when injected into the rat magnocellular nucleus basalis (MBN), impels excitotoxic injury of cholinergic projection neurons. Whereas learning and memory dysfunction is a hallmark of A beta-induced cholinergic deficits, anxiety, or hypoactivity under novel conditions cannot be attributed to the loss of cholinergic MBN neurons. As mood-related behavioral parameters are primarily influenced by the central serotonergic system, in the present study we investigated whether A beta(1-42) toxicity in the rat MBN leads to an altered serotonergic innervation pattern in the rat basal forebrain and cerebral cortex 7 days postsurgery. A beta infusion into the MBN elicited significant anxiety in the elevated plus maze. A beta toxicity on cholinergic MBN neurons, expressed as the loss of acetylcholinesterase-positive cortical projections, was accompanied by sprouting of serotonergic projection fibers in the MBN. In contrast, the loss of serotonin-positive fiber projections, decreased concentrations of both serotonin and 5-hydroxyindoleacetic acid, and decline of cortical 5-HT(1A) receptor binding sites indicated reduced serotonergic activity in the somatosensory cortex. In conclusion, the A beta-induced primary cholinergic deficit in the MBN and subsequent cortical cholinergic denervation bidirectionally modulate serotonergic parameters in the rat basal forebrain and cerebral cortex. We assume that enhanced serotonin immunoreactivity in the damaged MBN indicates intrinsic processes facilitating neuronal recovery and cellular repair mechanisms, while diminished cortical serotonergic activity correlates with the loss of the subcortical cholinergic input, thereby maintaining the balance of neurotransmitter concentrations in the cerebral cortex.

Effect of orthognathic surgery on the temporomandibular joint in patients with anterior open bite.

This study examined the prevalence of temporomandibular joint (TMJ) signs and symptoms in patients with anterior open bite. The influence of orthognathic surgery on the TMJ in these patients and the interaction of occlusal and psychologic variables on the presence and/or persistence of pain was studied. A retrospective survey of 83 patients with an anterior open bite who underwent orthognathic surgery was carried out. Records were examined for the prevalence of abnormal TMJ signs and symptoms, including pain. A survey was mailed to these patients that consisted of: (1) the TMJ Scale, (2) the Symptom Checklist 90 (SCL90), (3) the Spielberger State-Trait Anxiety Inventory (STAI), and (4) a visual analog scale on which patients indicated their degree of satisfaction with the procedure. Thirty-seven (42%) patients responded to the survey, and 13 (15%) also attended a clinical and radiographic examination. Multiple regression analysis was used for statistical analysis of the factors contributing to the presence and/or persistence of pain. In the preoperative group, the prevalence of pain was 32%, dysfunction 40%, and limitation of opening 7%. Age and gender were significantly associated with the presence of pain. The overall prevalence of abnormal TMJ signs and symptoms was not significantly different after orthognathic surgery. An abnormal psychologic profile was the most significant factor associated with the presence and/or persistence of pain. It is concluded that that the prevalence of temporomandibular disorders in anterior open bite patients increases with age, is significantly higher in females, and is not influenced by other occlusal variables. Furthermore, orthognathic surgery does not significantly influence temporomandibular disorders in patients with anterior open bite. Female patients, particularly those with an abnormal psychologic profile, are at a higher risk of persistent postoperative TMJ pain.

Prior exposure to methylenedioxyamphetamine (MDA) induces serotonergic loss and changes in spontaneous exploratory and amphetamine-induced behaviors in rats.

The substituted amphetamine 3,4 methylenedioxyamphetamine (MDA) is a popular recreational drug of abuse. Administration of MDA to experimental animals has been shown to induce damage to serotonergic axons and nerve terminals. However, there is a lack of information on whether these treatments can produce any long-term changes in behavioural performance particularly under stressful conditions. In the present study, MDA (7.5 mg/kg; i.p.) was administered twice daily to adult male Sprague Dawley rats for four days. Four weeks following the last dose, spontaneous behaviors of these animals were tracked and scored in a novel "open field" environment using an automated video registration and computer interpretation system. Changes in behavior were observed in MDA treated animals including reductions in exploratory oriented behaviors (locomotion and rearing) and increases in grooming behavior when compared to vehicle treated controls. MDA-treated animals also displayed an enhanced locomotor and stereotyped response to d-amphetamine (12 mg/kg; i.p.). Significant reductions in 5-HT concentrations (20-30%) were observed in the frontal cortex, amygdala, striatum, and hypothalamus as a result of MDA treatment. In addition, [3H] paroxetine binding was reduced by (40%) in the cortex of MDA treated rats indicating that the decrease in 5-HT concentrations were accompanied by a reduction in intact presynaptic 5-HT nerve terminals. Changes in behavioural performance in a novel "open field" environment and following d-amphetamine challenge might be considered as a behavioural model of serotonergic deficit induced by MDA. The findings of this study also suggest that MDA use may increase both the abuse potential, and the propensity to develop psychosis as a result of abusing other psychostimulants such as d-amphetamine.

Methylenedioxymethamphetamine-induced suppression of interleukin-1beta and tumour necrosis factor-alpha is not mediated by serotonin.

The purpose of the present study was to examine the role of serotonin release in methylenedioxymethamphetamine (MDMA)-induced immunosuppression in rats. We examined the effect of pretreatment with the selective serotonin reuptake inhibitor paroxetine, and the tryptophan hydroxylase inhibitor para-chlorophenylalanine on MDMA-induced suppression of interleukin-1beta and tumour necrosis factor (TNF)-alpha secretion following an in vivo lipopolysaccharide challenge. Although paroxetine blocked MDMA-induced serotonin depletion in the cortex and hypothalamus, it failed to alter the suppressive effect of MDMA on lipopolysaccharide-induced TNF-alpha secretion. Similarly, although para-chlorophenylalanine caused a 90% depletion in cortical and hypothalamic serotonin content, it failed to alter the suppressive effect of MDMA on lipopolysaccharide-induced interleukin-1beta or TNF-alpha secretion. In conclusion, although MDMA is a potent releaser of serotonin, the suppressive effects of MDMA on lipopolysaccharide-induced proinflammatory cytokine secretion cannot be attributed to its serotonin-releasing properties.

Behavioural, hyperthermic and neurotoxic effects of 3,4-methylenedioxymethamphetamine analogues in the Wistar rat.

1. The ability of N-ethyl (MDEA) and N-butyl (MDBA) analogues of 3,4-methylenedioxymethamphetamine (MDMA, 'Ecstasy') to induce acute behavioural changes and increases in body temperature, and to cause serotonergic neurotoxicity, was assessed in young adult male Wistar rats. The in vitro ability of MDMA analogues to evoke presynaptic monoamine release from crude rat forebrain synaptosomal preparations pre-labelled with [3H]5-HT or [3H]DA was also measured. 2. In behavioural experiments, acute MDMA and MDEA (20 mg/kg, i.p.) significantly increased rat open-field locomotion scores, decreased open-field rearing, and induced stereotypy, Straub tail and head weaving. MDBA did not produce any of these behaviours. 3. After repeated dosing (8 x 20 mg/kg, i.p., twice daily for 4 days), MDMA > MDEA >> MDBA > or = saline at decreasing forebrain [3H]paroxetine binding levels and concentrations of 5-HT and 5-HIAA at 14 days post-treatment. None of the analogues caused any long-term changes in dopamine or noradrenaline concentrations in the forebrain. 4. Acute MDMA and MDEA (20 mg/kg, i.p.) produced significant acute increases in rat aural temperature compared with saline-treated animals, while 20 mg/kg MDBA caused no significant effects. 5. MDA, MDMA and MDEA were equipotent at inducing [3H]5-HT release from frontal cortex/hippocampal synaptosomes, while MDBA only evoked a significant release at 100 microM concentrations. The potency order for inducing [3H]DA release from striatal synaptosomes was MDA > MDMA > MDEA = MDBA. 6. This study shows that large N-alkyl substitution decreases the ability of MDMA analogues to evoke presynaptic 5-HT and DA release, induce acute hyperthermia, hyperlocomotion and behavioural changes, and cause long-term serotonergic neurotoxicity. 7. The structure-activity relationship data presented here indicate that the neurotoxic damage caused by substituted amphetamines requires a combination of acute hyperthermia and increased neurotransmitter release. Induction of one of these effects in isolation is not sufficient to cause serotonergic nerve terminal degradation.