Altered neurochemical parameters in the brains of mice (Frings) susceptible to audiogenic seizures

In this study, several biochemical aspects of serotonergic neurons were compared in mice susceptible to audiogenic seizures (Frings) and in mice not susceptible to audiogenic seizures (CF 1). Tryptophan hydroxylase (TPH) activity was significantly lower in brains of Frings mice compared to CF1 mice; however, brain levels of 5-HT were similar in both strains. The significantly lower TPH activity in Frings mice was due to the altered kinetic characteristics of TPH activity in this strain. TPH from Frings mice had a significantly lower apparent maximal velocity and a significantly higher affinity for the substrate, as indicated by a lower apparent Km for tryptophan. The uptake of tryptophan and accumulation of 5-HT (following pargyline) were similar in Frings and CF 1 mice. The levels of 5-HT in selected brain regions of Frings and CF 1 mice were also similar, but NE levels were higher in the cerebral cortex and DA levels were higher in the neostriatum of Frings mice. It is unclear at this point what role, if any, 5-HT plays in susceptibility to audiogenic seizures in brains of Frings mice.     

B and T cells in the brains of autoimmune mice

Systemic lupus erythematosus (SLE) is an autoimmune disorder that can involve the central nervous system (CNS). Recently, we reported the presence of autoantibodies bound to the brain tissue of murine models of lupus; MRL/lpr and BXSB. We postulated that the source of these autoantibodies was in part due to in situ production, caused by the entry of B and T cells. Frozen brain sections of MRL/lpr and BXSB at 1 and 4 months of age were stained for CD3 (T cells) and CD19 (B cells) markers using an immunofluorescent antibody binding assay. Confocal fluorescence microscopy showed both CD3(+) and CD19(+) cells at 4 months of age only in MRL/lpr mice. There were no lymphocytes seen in the other autoimmune model, BXSB. Results suggest a difference in the mechanisms by which autoantibodies access the brain in these two autoimmune models of lupus.     

Increased TUNEL staining in brains of autoimmune Fas-deficient mice

Profound changes in brain morphology and behavior coincide with the spontaneous development of systemic autoimmune/inflammatory disease in Fas-deficient MRL-lpr mice. The dendrites atrophy, the density of hippocampal and cortical neurons decreases, and an anxious/depressive-like behavior emerges while lymphoid cells infiltrate into the choroid plexus of MRL-lpr mice. We hypothesized that the inherited lack of the Fas-dependent anti-inflammatory mechanism would lead to unsuppressed immune activity, characterized by reduced apoptosis in the MRL-lpr brain. Using the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeled (TUNEL) method as an indicator of apoptosis, a surprisingly high incidence of TUNEL-positive cells was observed in the hippocampus, choroid plexus and periventricular regions of MRL-lpr mice, 5-10-fold higher than that found in the MRL +/+ control brain. Immunostaining with anti-CD3, CD4 and CD8 monoclonal antibodies showed limited overlap between CD-positive and TUNEL-positive cells, suggesting that the dying cells are for the most part (approximately 70%) not T-lymphocytes. Although further characterization of the phenotype of the dying cells and the mechanism of cell death are required, the present results suggest the involvement of a Fas-independent apoptotic process in neurodegeneration induced by systemic autoimmune disease.     

Altered GABAergic neurotransmission in mice lacking dopamine D2 receptors

The levels of glutamic acid decarboxylase (GAD) were strongly increased in the cortex and the striatum in dopamine D2 receptors null (D2R-/-) mice, which show a significant locomotor impairment. In this study, the effects of different GABAergic drugs on locomotor activity were analyzed in D2R-/- mice. After administering muscimol (1 mg/kg), a GABA(A) receptor agonist, the D2R-/- mice showed increased locomotor activity up to 200%. When the muscimol dose was increased (4-6 mg/kg), the D2R-/- mice exhibited seizure-like behavior, and the electroencephalographic (EEG) recordings during these behaviors showed a high amplitude rhythmic epileptiform activity in these mice. In situ hybridization showed that after injecting muscimol in the D2R-/- mice, the expression of enkephalin and immediate early gene, NGFI-A, was closely regulated with the locomotor activity regulated by GABAergic stimulation. These results suggest that the absence of D2R alters the GABAergic neurotransmission, specifically on GABA(A)-receptor mediated signaling, and stimulating the GABA(A) receptor can reverse the dysfunction of GABAergic inhibition in the motor circuits in the basal ganglia.     

Modeling socially anhedonic syndromes: genetic and pharmacological manipulation of opioid neurotransmission in mice

Social anhedonia, or the diminished capacity to experience pleasure and reward from social affiliation, is a major symptom of different psychiatric disorders, including some forms of infantile autism and schizophrenia spectrum disorders. The brain opioid hypothesis of social attachment is a promising model for achieving insights into how neurobiological and developmental factors contribute to the regulation of social reward. In this study, genetic knocking-out and naltrexone (NTRX) treatment during the first 4 days of life were used to disrupt opioid neurotransmission in mouse pups and their attachment relationships with the mother. Both permanent (genetic) and transient (pharmacological) manipulations of opioid neurotransmission exerted long-term effects on social affiliation. When juveniles, both μ-opioid receptor knockout mice and NTRX-treated pups showed reduced interest in peers and no preference for socially rewarding environment. These results demonstrate that sociability in juvenile mice is highly dependent on the establishment during infancy of a positive affective relationship with their mothers and that opioid neurotransmission has a major role in the regulation of social hedonic capacity. If the validity of this animal model will be confirmed by future research, translational studies focusing on the interaction between early experience and opioid neurotransmission could provide useful insights for identifying endophenotypes of human psychiatric disorders associated with social anhedonia.     

Altered neurochemical markers in Rett's syndrome.

Rett's syndrome (RS) is a neurologic disorder associated with severe mental deficiency and neurologic manifestations of cortical and extrapyramidal origin. The present report is a preliminary postmortem brain study that compares the levels of endogenous biogenic amines and selected neurotransmitter receptors in five cases with RS and six normal controls of similar age. The level of choline acetyltransferase activity was reduced in several cortical and subcortical regions. Endogenous levels of dopamine in the superior frontal and superior temporal gyri, occipital cortex, and putamen were reduced. The changes in specific neurotransmitter markers, particularly those associated with the basal ganglia and cortex, may underlie the progressive deterioration in motor and cognitive function characteristic of this progressive disorder.     

Altered metabolic and neurochemical responses to chronic unpredictable stressors in ghrelin receptor‐deficient mice

Ghrelin, a hormone produced by the stomach, is generally associated with feeding responses and the regulation of food intake. Recent evidence, however, suggests that ghrelin is also a stress hormone, given that it is released following acute and chronic stressors. The present study examined the role of ghrelin in producing normal metabolic and neurochemical responses to chronic stress. This was achieved by examining these responses in mice with targeted deletions of the ghrelin receptor gene (GHSR KO mice), and comparing them with the same responses in their wild‐type (WT) littermates. As expected, WT stressed mice decreased their caloric intake, body weight gain and caloric efficiency while maintaining adiposity. GHSR KO mice, however, did not show these alterations despite having normal glucocorticoid responses to stress. In parallel with these changes, chronic unpredictable stress caused changes in norepinephrine, dopamine and serotonin in a number of brain regions. Of these, norepinephrine neurotransmission in the arcuate nucleus and prefrontal cortex was differentially altered in GHSR KO mice. Within the nucleus acumbens, dopamine utilization was increased in WT mice but not in GHSR KO mice. Finally, there were strain differences in serotonin neurotransmission that may explain interstrain body weight and adiposity differences. These results suggest that the metabolic changes necessary to deal with the energetic challenge presented by repeated exposure to stressors do not occur in GHSR KO mice, and they are discussed within the context of the potential vulnerability to stress‐induced pathology.     

Altered GABAergic neurotransmission is associated with increased kainate-induced seizure in prostaglandin-endoperoxide synthase-2 deficient mice

Excitotoxicity involves over activation of brain excitatory glutamate receptors and has been implicated in neurological, neurodegenerative and neuropsychiatric diseases. Metabolism of arachidonic acid (AA) through the phospholipase A(2) (PLA(2))/prostaglandin-endoperoxide synthase (PTGS) pathway is increased after excitotoxic stimulation. However, the individual roles of the PTGS isoforms in this process are not well established. We assessed the role of the PTGS isoforms in the process of excitotoxicity by exposing mice deficient in either PTGS-1 (PTGS-1(-/-)) or PTGS-2 (PTGS-2(-/-)) to the prototypic excitotoxin, kainic acid (KA). Seizure intensity and neuronal damage were significantly elevated in KA-exposed PTGS-2(-/-), but not in PTGS-1(-/-), mice. The increased susceptibility was not associated with an alteration in KA receptor binding activity or mediated through the CB1 endocannabinoid receptor. The frequency of spontaneous inhibitory postsynaptic currents (sIPSCs) was decreased in the CA1 pyramidal neurons of PTGS-2(-/-) mice, suggesting an alteration of GABAergic function. In wild-type mice, six weeks treatment with the PTGS-2 selective inhibitor celecoxib recapitulated the increased susceptibility to KA-induced excitotoxicity observed in PTGS-2(-/-) mice, further supporting the role of PTGS-2 in the excitotoxic process. The increased susceptibility to KA was also associated with decreased brain levels of PGE(2), a biomarker of PTGS-2 activity. Our results suggest that PTGS-2 activity and its specific products may modulate neuronal excitability by affecting GABAergic neurotransmission. Further, inhibition of PTGS-2, but not PTGS-1, may increase the susceptibility to seizures.     

Altered depression-related behavior and neurochemical changes in serotonergic neurons in mutant R406W human tau transgenic mice

Mutant R406W human tau was originally identified in frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17) and causes a hereditary tauopathy that clinically resembles Alzheimer's disease (AD). In the current study, we examined the performance of R406W transgenic (Tg) mice in the forced swimming test, a test with high predictivity of antidepressant efficacy in human depression, and found an enhancement of the immobility time. In contrast, the motor function and anxiety-related emotional response of R406W Tg mice were normal. Furthermore, a selective serotonin reuptake inhibitor (SSRI), fluvoxamine (100 mg/kg, p.o.), significantly reduced this enhancement of the immobility time, whereas a noradrenaline reuptake inhibitor, desipramine, had no effect. In an in vivo microdialysis study, R406W Tg mice exhibited a significantly decreased extracellular 5-hydroxyindoleacetic acid (5-HIAA) level in the frontal cortex and also exhibited a tendency toward a decreased extracellular 5-hydroxytryptamine (5-HT) level. Moreover, fluvoxamine, which reduced the enhancement of the immobility time, significantly increased the extracellular 5-HT level in R406W Tg mice. These results suggest that R406W Tg mice exhibit changes in depression-related behavior involving serotonergic neurons and provide an animal model for investigating AD with depression.     

Presynaptic receptors and modulation of neurotransmission: pharmacological implications and therapeutic relevance

During noradrenaline (NA) release elicited by the arrival of nerve impulses, the neurotransmitter interacts with specific receptors (α, β₁ or β₂-adrenoceptors) located in the membrane of the postsynaptic cell to trigger responses in the effector organ. Until a few years ago, the role o f noradrenergic nerve terminals in neurotransmission was considered to be concerned exclusively with the synthesis, storage, release, and inactivation of NA and there were no indications that receptors might also be present in the Presynaptic membrane. However, it is now clear that a number of Presynaptic receptors exist which modulate NA release. Here S. Z. Langer reviews their separate pharmacological properties and discusses the therapeutic relevance of these new findings.     

Altered behavioral and neurochemical response to stress in benzodiazepine-withdrawn rats

Rats chronically treated with diazepam (DZM) (2 mg/kg/day i.p.) for 21 days, were subsequently tested at 24, 48, 72 and 96 h after the last injection in the elevated plus-maze. When compared with their respective controls, withdrawn animals showed an anxiogenic response since they exhibited a significant decrease in the % of time spent in the open arms, and a reduction in open and total arms entries at 24 and 48 h following the last benzodiazepine administration. No other behavioral differences were observed in the remaining groups. Ninety-six hours following DZM withdrawal, when the anxiogenic response was no longer evident, withdrawn rats showed a significant decrease in immobility time when exposed to a forced-swim test (FST). GABA stimulation of chloride uptake in cortical tissue was measured in BDZ-withdrawn rats or vehicle-treated animals with or without exposure to the FST. An enhanced chloride uptake following GABA stimulation was observed in vehicle-treated rats following the swimming trial. However, similar values of chloride uptake were found among rats withdrawn from DZM at 96 h either exposed or not to the FST and vehicle-treated animals without prior stress exposure. These findings show that BDZ withdrawal alters the neurochemical and behavioral response to a subsequent stressful experience. These lines of evidence may indicate that BZD withdrawal reduced the ability to develop an adaptive response to stress.     

Altered GABA neurotransmission and prefrontal cortical dysfunction in schizophrenia.

Dysfunction of the dorsolateral prefrontal cortex appears to be a central feature of the pathophysiology of schizophrenia, and this dysfunction may be related to alterations in gamma aminobutyric acid (GABA) neurotransmission. Determining the causes and consequences of altered GABA neurotransmission in schizophrenia, and the relationship of these changes to other abnormalities in prefrontal cortical circuitry, requires an understanding of which of the multiple subpopulations of cortical GABA neurons are affected. The chandelier class of GABA neurons, especially those located in the middle layers of the prefrontal cortex (PFC), have been hypothesized to be preferentially involved in schizophrenia because they 1) receive direct synaptic input from dopamine axons, 2) exert powerful inhibitory control over the excitatory output of layer 3 pyramidal neurons, and 3) undergo substantial developmental changes during late adolescence, the typical age of onset of schizophrenia. Consistent with this hypothesis, the axon terminals of chandelier neurons, as revealed by immunoreactivity for the GABA membrane transporter, are reduced substantially in the middle layers of the PFC in schizophrenic subjects. This alteration appears to be selective for the chandelier class of GABA neurons and for the disease process of schizophrenia. These findings provide insight into the pathophysiologic mechanisms underlying prefrontal cortical dysfunction in schizophrenia, and they reveal new targets for therapeutic intervention in this illness.     

Behavioral and neurochemical consequences of lipopolysaccharide in mice: anxiogenic-like effects

Systemic administration of lipopolysaccharide (LPS) induces sickness behaviors, as well as alterations of hypothalamic-pituitary-adrenal functioning commonly associated with stressors. In the present investigation, it was demonstrated that systemic LPS treatment induced a sickness-like behavioral profile (reduced active behaviors, soporific effects, piloerection, ptosis), which appeared to be dependent upon the novelty of the environmental context in which animals were tested. As well, LPS induced anxiogenic-like responses, including decreased time spent in the illuminated portion of a light-dark box, reduced open-arm entries in a plus-maze test, and decreased contact with a novel stimulus object in an open-field situation. The behavioral changes were accompanied by increased plasma ACTH and corticosterone levels. As well, LPS induced increased turnover of norepinephrine (NE), dopamine (DA) and serotonin (5-HT) in the paraventricular nucleus (PVN), median eminence plus arcuate nucleus, hippocampus, as well as NE turnover within the locus coeruleus and DA turnover within the nucleus accumbens. Although these neurochemical variations were reminiscent of those elicited by stressors, LPS was not particularly effective in modifying DA activity within the prefrontal cortex or NE within the amygdala, variations readily induced by stressors. Whether the LPS-induced anxiogenic-like responses were secondary to the illness engendered by the endotoxin remains to be determined. Nevertheless, it ought to be considered that bacterial endotoxin challenge, and the ensuing cytokine changes, may contribute to emotionality and perhaps even anxiety-related behavioral disturbances.     

Altered angioarchitecture in selected areas of brains with Alzheimer's disease

Summary It was the aim of this study to determine, qualitatively and quantitatively, alterations in the blood vessels of brains removed postmortem from patients with Alzheimer's disease (AD), and to compare these findings with the appearance of cerebral blood vessels in a group of individuals without brain disorders. Celloidin sections of brain tissue from four cerebral areas, pre-frontal (Brodmann's area 9), basal forebrain, sensorimotor, and hippocampus, were subjected to an alkaline phosphatase reaction to facilitate the evaluation of the vascular distribution. The vascular density in five sections was determined by counting the number of vascular intersections with a microscopic test grid of 100 squares; ten fields per section were examined in this manner. Analysis of 16 AD and 6 control brains, showed that there was a striking and statistically significant reduction in the vascular net density specifically in the basal forebrain region and the hippocampus of AD brains. In addition, vessels in the AD brains exhibited extensive topographical changes, such as kinking and looping. These results indicate that modifications in vascular density are present in AD brains with a marked regional specificity.Supported in part by the St. Louis Alzheimer's Disease and Related Disorders Association and the Department of Community Medicine. St. Louis University Medical Center     

Altered behavioural and neurochemical profile of L-DOPA following deuterium substitutions in the molecule

In Parkinson's disease patients, a prolonged half-life of dopamine formed from l-DOPA may reduce the risk of developing l-DOPA-induced side-effects. Deuterium substitutions in the l-DOPA molecule are expected to yield dopamine with an altered half-life because C–D bonds are more stable than C–H bonds. Therefore we tested, in the rat, the neurochemical and behavioral effects of different types of l-DOPA with deuterium substitutions at the α-carbon and/or the β-carbon. By means of microdialysis, we found that l-DOPA with 3 deuterium substitutions (D3-l-DOPA) enhanced dopamine output in the striatum more effectively than l-DOPA and all the other deuterium variants. Moreover, D3-l-DOPA produced a more pronounced stimulation of locomotor activity in reserpinized rats compared to conventional l-DOPA. In contrast β,β-D2-l-DOPA was less effective than l-DOPA in raising striatal dopamine levels and was ineffective at restoring locomotor activity in reserpinized rats. These results demonstrate that the introduction of deuterium at different positions in the l-DOPA molecule dramatically changes its behavioral and neurochemical profile and suggest that l-DOPA treatment of Parkinson's disease may be improved in this way.     

Development of obesity and neurochemical backing in aurothioglucose-treated mice

To clarify the neurochemical backing of aurothioglucose (ATG)-induced obesity in mice, we investigated lesion sites, hypothalamic neurotransmitters and c-Fos-like immunoreactivity (Fos-IR). At day 2 after ATG, tissue loss or cells death was observed in several parts of the ventral area of the ventromedial hypothalamic nucleus (VMH), and the dorsal area of arcuate nucleus and in the nucleus of the solitary tract (NTS). However, the greater part of the VMH was retained. Body weight began to increase in week 1. Hypothalamic serotonin (5-HT) and the metabolites were increased at day 2. The contents of acetylcholine, norepinephrine and dopamine in the hypothalamus showed no significant change. In week 1, the area shown tissue loss was compacted and plugged up. In the control group, most obvious c-Fos-like immunoreactive region was paraventricular nucleus (PVN). At day 2, Fos-IR was observed around destroyed regions in the hypothalamus and NTS, but few Fos-IR was found in the other regions including PVN. The Fos-IR around destroyed regions diminished after week 1. In week 3, Fos-IR in the PVN increased. These results suggest that the development of ATG-induced obesity cannot be attributed to solely VMH destruction. The restoration processes of the neuronal dysfunction involving PVN seem to play an important role in the development of obesity. NTS lesion and 5-HT system might contribute to decrease in food intake for several days after ATG.     

Altered inflammatory response and increased neurodegeneration in metallothionein I+II deficient mice during experimental autoimmune encephalomyelitis

Metallothionein-I+II (MT-I+II) are antioxidant, neuroprotective proteins, and in this report we have examined their roles during experimental autoimmune encephalomyelitis (EAE) by comparing MT-I+II-knock-out (MTKO) and wild-type mice. We herewith show that EAE susceptibility is higher in MTKO mice relatively to wild-type mice, and that the inflammatory responses elicited by EAE in the central nervous system (CNS) are significantly altered by MT-I+II deficiency. Thus, during EAE the MTKO mice showed increased macrophage and T-lymphocytes infiltration in the CNS, while their reactive astrogliosis was significantly decreased. In addition, the expression of the proinflammatory cytokines interleukin-1beta, interleukin-6, and tumor necrosis factor-alpha elicited by EAE was further increased in the MTKO mice, and oxidative stress and apoptosis were also significantly increased in MTKO mice compared to normal mice. The present results strongly suggest that MT-I+II are major factors involved in the inflammatory response of the CNS during EAE and that they play a neuroprotective role in this scenario.     

Presynaptic parkinsonism in olivopontocerebellar atrophy: clinical, pathological, and neurochemical evidence

The substrate for olivopontocerebellar atrophy parkinsonism is obscure due to the lack of clinical and pathological reports and the absence of studies on dopamine receptors in this entity. We describe a patient with olivopontocerebellar atrophy whose clinical presentation was levodopa-responsive parkinsonism in whom pathological examination disclosed pronounced nigral cell loss with no striatal damage. Autoradiographic labeling with 3H-spiperone showed normal densities of D2 dopamine striatal receptors. These data show that indistinguishable nigral, presynaptic parkinsonism occurs in patients with idiopathic Parkinson's disease and in patients with olivopontocerebellar atrophy, and also how a favorable response to levodopa is neither synonymous with idiopathic Parkinson's disease, nor does it exclude multiple-system, atrophy-related parkinsonism.     

Altered norepinephrine regulation in bulimia: effects of pharmacological challenge with isoproterenol

While abnormalities in central norepinephrine regulation may contribute to abnormal eating patterns in bulimia nervosa, alterations in function of the peripheral sympathetic nervous system could contribute to the decreased metabolic rate and increased anxiety responses previously reported in these patients. To assess beta-adrenergic receptor sensitivity in bulimic patients, we studied cardiovascular and hormonal responses to acute pharmacological challenge with intravenously administered isoproterenol. In comparison to healthy controls, binge-abstinent bulimic patients had significantly reduced mean baseline plasma norepinephrine level, pulse rate, and systolic blood pressure, and significantly increased chronotropic responses to isoproterenol infusion. Decreased sympathoneural activity may contribute to a tendency for bulimic patients to maintain body weight despite low caloric intake.