Methamphetamine and human immunodeficiency virus protein Tat synergize to destroy dopaminergic terminals in the rat striatum.

Dysfunction of the dopaminergic system accompanied by loss of dopamine in the striatum is a major feature of human immunodeficiency virus-1-associated dementia. Previous studies have shown that human immunodeficiency virus-1-associated dementia patients with a history of drug abuse have rapid neurological progression, prominent psychomotor slowing, more severe encephalitis and more severe dendritic and neuronal damage in the frontal cortex compared with human immunodeficiency virus-1-associated dementia patients without a history of drug abuse. In a previous study, we showed that methamphetamine and human immunodeficiency virus-1 protein Tat interact to produce a synergistic decline in dopamine levels in the rat striatum. The present study was carried out to understand the underlying cause for the loss of dopamine. Male Sprague-Dawley rats were administered saline, methamphetamine, Tat or Tat followed by methamphetamine 24 h later. Two and seven days later the animals were killed and tissue sections from striatum were processed for silver staining to examine terminal degeneration while sections from striatum and substantia nigra were processed for tyrosine hydroxylase immunoreactivity. Striatal tissue was also analyzed by Western blotting for tyrosine hydroxylase protein levels. Compared with controls, methamphetamine+Tat-treated animals showed extensive silver staining and loss of tyrosine hydroxylase immunoreactivity and protein levels in the ipsilateral striatum. There was no apparent loss of tyrosine hydroxylase in the substantia nigra. Markers for oxidative stress were significantly increased in striatal synaptosomes from Tat+methamphetamine group compared with controls. The results indicate that methamphetamine and Tat interact to produce an enhanced injury to dopaminergic nerve terminals in the striatum with sparing of the substantia nigra by a mechanism involving oxidative stress. These findings suggest a possible mode of interaction between methamphetamine and human immunodeficiency virus-1 infection to produce enhanced dopaminergic neurotoxicity in human immunodeficiency virus-1 infected/methamphetamine-abusing patients.

Neuronal injury in hippocampus with human immunodeficiency virus transactivating protein, Tat.

Patients with human immunodeficiency virus infection may develop a dementing illness. Using both in vitro and in vivo models, we investigated the susceptibility of the hippocampal formation to the Tat protein of human immunodeficiency virus. We also determined the pattern of hippocampal injury in patients with human immunodeficiency virus encephalitis. Following exposure of hippocampal slices to Tat, marked susceptibility of CA3 region with relative insensitivity of the CA1/2 region was observed. Injection of Tat into different regions of the rat hippocampus produced similar neuronal loss in both CA3 region and the dentate gyrus. In animals administered Tat, lesions were dose-dependent and immunohistochemical staining showed marked gliosis and loss of microtubule associated protein-2 in the affected areas at 3 days post-injection. Interestingly, synaptophysin staining was relatively preserved. In hippocampal tissue from patients with human immunodeficiency virus encephalitis, loss of microtubule-associated protein-2 staining was reduced in the molecular layer of the dentate gyrus. The results of our experiments demonstrate a unique pattern of hippocampal injury in organotypic culture and rats exposed to Tat. Our observations that patients with human immunodeficiency virus reveal a similar pattern of damage suggests that Tat protein may be pathophysiological relevant in human immunodeficiency virus encephalitis.

Acceleration of HIV dementia with methamphetamine and cocaine.

We report a patient with rapidly accelerating HIV dementia accompanied by seizures and an unusual movement disorder despite highly potent antiretroviral therapy. This clinical constellation was associated with the non-parenteral use of methamphetamine and cocaine. Fractional enhancement time on post contrast magnetic resonance imaging studies revealed a progressive breakdown of the blood brain barrier particularly in the basal ganglia. The movement disorder but not the dementia responded to a combination of dopamine replacement and anticholinergic therapy. While the movement disorder may have been unmasked by concomitant anticonvulsant therapy, we suggest in this instance, that prior drug abuse synergized with HIV to cause a domino effect on cerebral function. Careful attention and analysis to histories of remote non-injecting drug abuse may help substantiate our hypothesis.

Dopamine induces proteasome inhibition in neural PC12 cell line.

The autoxidation and enzymatic catabolism of dopamine results in the generation of reactive oxygen species (ROS), which may possibly contribute to oxidative stress in multiple neurodegenerative disorders. Recent studies indicate that proteasome inhibition occurs in numerous neurodegenerative conditions, possibly as the result of oxidative stress, although the effects of dopamine on proteasome activity have not been determined. In the present study we examined the effects of dopamine on proteasome activity in the neural PC12 cell line. Application of dopamine induced a dose- and time-dependent decrease in proteasome activity, which occurred prior to cell death. Application of an antioxidant (gluthathione monoethyl ester), monoamine oxidase inhibitors (deprenyl, clogyline, paragyline), or an inhibitor of dopamine uptake (nomifensine) attenuated dopamine toxicity and dopamine-induced proteasome impairment. Application of the proteasome inhibitor lactacystin increased the toxicity of dopamine and the levels of protein oxidation following administration of dopamine. Together, these data indicate that dopamine induces proteasome inhibition that is dependent, in part, on ROS and dopamine uptake, and suggest a possible role for proteasome inhibition in dopamine toxicity.

Methamphetamine toxicity is attenuated in mice that overexpress human manganese superoxide dismutase.

We have investigated methamphetamine (MA) toxicity in transgenic mice that overexpress the human form of mitochondrial manganese superoxide dismutase (MnSOD). Our results reveal a significant reduction in the long-term depletion of striatal dopamine and protein oxidation following repeated administration of MA in transgenic vs. non-transgenic littermates. These findings support the notion that ROS contribute to MA-induced brain damage and suggest that mitochondria may play an important role in this form of neurodegeneration.

Neuronal cell death in Huntington's disease: a potential role for dopamine.

Huntington's disease is an inherited neurodegenerative disorder, the cause of which is unknown. Excitotoxicity, mitochondrial dysfunction and oxidative stress are all likely to contribute to the striatal cell death that occurs in this disorder. There are accumulating data indicating that under specific circumstances, dopamine, which occurs in high concentrations in the basal ganglia, might be neurotoxic. In this article, the current models used to study Huntington's disease are reviewed and the recent findings that implicate dopamine in the pathophysiology of this progressive disorder are discussed. Although many questions remain unanswered, the dopaminergic system could contribute to striatal vulnerability in Huntington's disease and provide a novel avenue for the development of new therapies.

Evidence that Par-4 participates in the pathogenesis of HIV encephalitis.

Progressive neuronal degeneration in brain regions involved in learning and memory processes is a common occurrence in patients infected with human immunodeficiency virus type 1 (HIV-1). We now report that levels of Par-4, a protein recently linked to neuronal apoptosis in Alzheimer's disease, are increased in neurons in hippocampus of human patients with HIV encephalitis and in monkeys infected with a chimeric strain of HIV-1 and simian immunodeficiency virus. Par-4 levels increased rapidly in cultured hippocampal neurons following exposure to the neurotoxic HIV-1 protein Tat, and treatment of the cultures with a Par-4 antisense oligonucleotide protected the neurons against Tat-induced apoptosis. Additional findings show that Par-4 participates at an early stage of Tat-induced neuronal apoptosis before caspase activation, oxidative stress, and mitochondrial dysfunction. Our data suggest that Par-4 may be a mediator of neuronal apoptosis in HIV encephalitis and that therapeutic approaches targeting the Par-4 apoptotic cascade may prove beneficial in preventing neuronal degeneration and associated dementia in patients infected with HIV-1.

Clorgyline and deprenyl attenuate striatal malonate and 3-nitropropionic acid lesions.

We have previously shown that dopamine depletion reduces striatal damage elicited by the mitochondrial neurotoxins malonate and 3-nitropropionic acid (3NP). Metabolism of dopamine by monoamine oxidase results in the formation of hydrogen peroxide, which may mediate dopamine toxicity. In this study, administration of the monoamine oxidase inhibitors clorgyline and deprenyl resulted in a 42% and 75% reduction in lesion volumes in malonate- and 3NP-treated animals, respectively, compared to controls.

6-Hydroxydopamine injections into the nigrostriatal pathway attenuate striatal malonate and 3-nitropropionic acid lesions.

The mitochondrial inhibitors malonate and 3-nitropropionic (3NP) acid are potent neurotoxins in vivo. Administration of these compounds results in neuronal loss similar to that seen in Huntington's disease. Although the mechanism of cell death produced by these compounds likely involves activation of N-methyl-D-aspartate receptors, it remains unclear why the striatum demonstrates regional susceptibility to the toxicity of these and other mitochondrial poisons. We hypothesized that dopamine, a weak neurotoxin that occurs in high concentrations in the striatum, may contribute to the neuronal damage caused by mitochondrial inhibition. We investigated whether depletion of striatal dopamine using the catecholaminergic toxin 6-hydroxydopamine would attenuate lesions induced by mitochondrial inhibition. We found that dopamine depletion reduced significantly the extent of histological damage in the striatum elicited by both intraparenchymal injections of 0.8 micromol malonate and 20 mg/kg systemic administration of 3NP. These data suggest that dopamine or one of its metabolites may contribute to mitochondrial toxin-induced cell death.

The mitochondrial inhibitor malonate enhances NMDA toxicity in the neonatal rat striatum.

Intra-striatal injections of the mitochondrial inhibitor malonate elicit age-dependent neuronal damage in rat brain; injury is more extensive in older animals than in young adults. We investigated the neurotoxic potential of malonate in the immature rat brain. We found that 7-day-old (P7) rats were highly resistant to malonate neurotoxicity. Yet, although intra-striatal injections of 1 mumol malonate did not elicit overt tissue injury in P7 rats, co-administration of this dose of malonate with a dose of NMDA close to its toxicity threshold (2.5 nmol) doubled the severity of resulting excitotoxic injury.

Inhibition of nitric oxide synthase activity attenuates striatal malonate lesions in rats.

Mitochondrial inhibitors such as malonate are potent neurotoxins in vivo. Intrastriatal injections of malonate result in neuronal damage reminiscent of "excitotoxic" lesions produced by compounds that activate NMDA receptors. Although the mechanism of cell death produced by malonate is uncertain, overactivation of NMDA receptors may be involved; pretreatment of animals with NMDA antagonists provides neuroprotection against malonate lesions. NMDA receptor activation stimulates the enzyme nitric oxide (NO) synthase (NOS). Elevated tissue levels of NO may generate highly reactive intermediates that impair mitochondrial function. We hypothesized that NO may be a mediator of malonate toxicity. We investigated whether in vivo inhibition of NO production by the NOS inhibitor N omega-nitro-L-arginine (NLA) would attenuate lesions produced by intrastriatal injections of malonate. We found that systemic injections of 3 mg/kg of NLA significantly reduced the extent of histologic damage elicited by intrastriatal injections of 1.5 mumol of malonate in adult rats.

Resistance to nitroprusside neurotoxicity in perinatal rat brain.

Nitric oxide (NO) may mediate some of the toxic effects of the excitatory amino acid (EAA) glutamate when there is overactivation of the N-methyl-D-aspartate (NMDA) receptor. In the developing rodent nervous system, NMDA neurotoxicity peaks at postnatal day 7. To assess whether NO toxicity exhibits a similar developmental profile, we injected the NO generator sodium nitroprusside into the immature and adult rodent hippocampal formation and striatum, using a dose known to damage the adult nervous system. Contrary to our expectations, we found the immature brain highly resistant to the toxic effects of sodium nitroprusside.

Neurotransmitter receptors in Alzheimer disease.

Alzheimer disease (AD) is an exceedingly complex disorder in which numerous populations of neurons and neurotransmitter systems are damaged or destroyed. Effective treatment of the cognitive symptoms of AD does not exist, and new targets for therapeutic intervention are needed desperately. Traditionally, the neurotransmitter receptors have been the focus of new neuropsychopharmacological agents, so it seems reasonable to assess the status of these receptors in the AD brain. In this article, we review the quarter century of receptor research in AD. The limitations of receptor studies, in general, and the particular limitations of studying AD tissue are discussed. The cholinergic and glutamatergic systems have been implicated most directly in normal cognitive function, so the receptors for these neurotransmitters are emphasized in this review. We have attempted to point out the possible neurobiological roles and potential clinical significance of the various receptors in AD. Investigation of neurotransmitter receptors in AD provides a rational approach to the development of therapeutic and diagnostic strategies, and, at a minimum, should lead to a better understanding of the neurobiology of AD.

A study of cortical and hippocampal NMDA and PCP receptors following selective cortical and subcortical lesions.

The neuronal localization of glutamate and phencyclidine (PCP) receptors was evaluated in the cerebral cortex and hippocampal formation of rat CNS using quantitative autoradiography. Scatchard analysis of [3H]glutamate binding in the cortex (layers I and II and V and VI) showed no difference in the total number of binding sites (Bmax) or apparent affinity (Kd) 1 week, 1 month and 2 months following unilateral ibotenate lesions to nucleus basalis of Meynert (nbM) compared to the non-lesioned side. Quisqualic acid displacement of [3H]glutamate in layers I and II, 1 week following nbM destruction, revealed both high- and low-affinity binding sites (representing the quisqualate (QA) and N-methyl-D-aspartate (NMDA) sites, respectively). Compared to the control side, there was no difference in binding parameters for either of the receptor sites. In similarly lesioned animals, the NMDA receptor was specifically labelled with [3H]glutamate and the associated PCP receptor labelled with [3H]N-(1-[2-thienyl]cyclohexyl)3,4-piperidine ([3H]TCP) in adjacent brain sections. For both receptors, there was no change in the total number of binding sites in the cortex following destruction of nbM. On the other hand, virtually all binding to NMDA and PCP receptors was eliminated following chemical destruction of intrinsic cortical neurons. These results suggest that the NMDA/PCP receptor complex does not exist on the terminals of cortical cholinergic afferents. One week after knife cuts of the glutamatergic entorhinal pathway to the hippocampal formation only an approximate 10% reduction of NMDA and PCP receptors was seen in the dentate gyrus. Conversely, selective destruction of the dentate granule cells using colchicine caused a near identical loss of NMDA and PCP receptors (84% vs 92% respectively). It is concluded from these experiments that glutamate and PCP receptors exist almost exclusively on neurons intrinsic to the hippocampal formation and that no more than 10% of NMDA and PCP receptors exist as autoreceptors on glutamatergic terminals.

Excitatory amino acid binding sites in the hippocampal region of Alzheimer's disease and other dementias.

Quantitative receptor autoradiography was used to measure muscarinic cholinergic, benzodiazepine, kainate, phencyclidine (PCP), N-methyl-D-aspartate (NMDA) (measured in Tris acetate), quisqualate-sensitive, non-quisqualate-sensitive and total glutamate (measured in Tris chloride buffer) binding sites in adjacent sections of the hippocampal region of 10 Alzheimer's disease, nine control, and six demented, non-Alzheimer's disease postmortem human brains. The measurements were compared to the number of neurofibrillary tangles as revealed by Congo red staining of adjacent sections. All assays and measurements were done by observers blinded to the clinical diagnoses. Binding was decreased significantly for all ligands except quisqualate in stratum pyramidale of CA1 of the Alzheimer's disease brains. The binding loss was significantly greater for the non-quisqualate and NMDA sites than for the muscarinic, benzodiazepine and kainate sites with the total glutamate and PCP site losses being intermediate. Only the loss of benzodiazepine binding was significantly correlated with the number of neurofibrillary tangles. Lesser binding losses were seen in adjacent areas. This difference in the degree of binding decrease is consistent with the hypothesis that NMDA receptors are located on more distal dendrites of hippocampal neurons. There they may be relatively more vulnerable than the other receptors to the pathological process.

Quantitative autoradiographic localization of NMDA, quisqualate and PCP receptors in the frog tectum.

An organizing role for the N-methyl-D-aspartate (NMDA) receptor/channel has been suggested in the development of the retinotectal projection in Rana pipiens. The regional distributions of NMDA, phencyclidine (PCP) and quisqualic acid (QA) receptors were quantified using in vitro autoradiography in the tectum of normal and surgically produced 3-eyed juvenile frogs. NMDA and QA receptor binding was highest in the pretectum. Of the tectal layers, the superficial retinotectal synaptic zone, layer 9, had the highest amount of NMDA and QA receptor binding. Moderate binding was observed in layer 5, with little binding in the cellular layer 6. No specific [3H]N-(1-[2-thienyl]cyclohexyl) piperidine ([3H]TCP) binding was observed in any of the tectal regions.

Neurons of origin and fiber trajectory of amygdalofugal projections to the medial preoptic area in Syrian hamsters.

The amygdaloid neurons of origin and the trajectory of amygdaloid fibers to the medial preoptic area of the adult male Syrian hamster were identified by using horseradish peroxidase (HRP) histochemistry. After iontophoresis of HRP into the medial preoptic area, retrogradely labeled amygdaloid neurons were located in the dorsal and caudal parts of the medial amygdaloid nucleus and throughout the amygdalohippocampal area. No amygdaloid neurons were labeled after HRP applications confined to the most rostral portion of the medial preoptic area (anterior to the body of the anterior commissure). Following more caudal medial preoptic area injections (body of the anterior commissure to the suprachiasmatic nucleus) the distribution of retrogradely labeled cells in the medial amygdaloid nucleus and the amygdalohippocampal area revealed no topographic organization of the amygdalopreoptic connections. When amygdaloid neurons were labeled, the amygdalohippocampal area contained two to five times as many HRP-filled cells as the medial amygdaloid nucleus. Retrogradely transported HRP could be followed from the medial preoptic area to the amygdala through fibers in the dorsomedial quadrant of the stria terminalis. In addition, electrolytic lesions of the stria terminalis prior to iontophoresis of HRP into the medial preoptic area prevented retrograde transport to neurons in both the dorsocaudal medial amygdaloid nucleus and the amygdalohippocampal area. These results confirm earlier observations describing the location of autoradiographically labeled efferents from the medial amygdaloid nucleus to the medial preoptic area and provide new information about the restricted region within the medial amygdaloid nucleus from which these projections arise. They also suggest that, unlike the projections from the medial amygdaloid nucleus to the bed nucleus of the stria terminalis, the efferents to the medial preoptic area travel entirely in the stria terminalis.

Anatomic correlation of NMDA and 3H-TCP-labeled receptors in rat brain.

Using quantitative autoradiography, we have compared the regional distribution of N-methyl-D-aspartate (NMDA) receptors labeled with 3H-glutamate and dissociative anesthetic binding sites labeled with 3H-N-(1-[2-thienyl]cyclohexyl)3,4-piperidine (3H-TCP). Binding of both ligands was highest in the hippocampal formation, with high concentrations in a number of cortical and olfactory regions. Intermediate amounts of binding for both ligands were measured in several thalamic and basal telencephalic structures. Very little binding was observed in the hypothalamus, some deep forebrain regions, and most brain-stem structures. Linear-regression analysis comparing the binding at both sites revealed a marked concordance (R = 0.95; p less than 0.001; Pearson product-moment). The granule cell layer of the cerebellum was the only region in which this concordance was not observed. Scatchard analysis of 3H-glutamate binding to NMDA receptors in stratum radiatum of hippocampal formation revealed an apparent single binding site with a Bmax of 9.78 +/- 0.84 pmol/mg protein and KD of 158 +/- 37 nM. 3H-TCP also bound to an apparent single site with a Bmax of 2.07 +/- 0.16 pmol/mg protein and KD of 127 +/- 30 nM. Our results are consistent with the hypothesis that the dissociative anesthetic binding site is linked to the NMDA receptor, and the data suggest that there are approximately 4-5 NMDA binding sites for each dissociative anesthetic binding site.