20 results on '"Low MJ"'
Search Results
2. The Homeodomain Transcription Factor NKX2.1 Is Essential for the Early Specification of Melanocortin Neuron Identity and Activates Pomc Expression in the Developing Hypothalamus.
- Author
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Orquera DP, Tavella MB, de Souza FSJ, Nasif S, Low MJ, and Rubinstein M
- Subjects
- Animals, Body Weight physiology, Female, Gene Expression Regulation, Developmental physiology, Hypothalamus embryology, Hypothalamus metabolism, Male, Mice, Mice, Inbred C57BL, Mice, Transgenic, Melanocortins metabolism, Neurogenesis physiology, Neurons metabolism, Pro-Opiomelanocortin metabolism, Thyroid Nuclear Factor 1 metabolism
- Abstract
Food intake is tightly regulated by a group of neurons present in the arcuate nucleus of the hypothalamus, which release Pomc -encoded melanocortins, the absence of which induces marked hyperphagia and early-onset obesity. Although the relevance of hypothalamic POMC neurons in the regulation of body weight and energy balance is well appreciated, little is known about the transcription factors that establish the melanocortin neuron identity during brain development and its phenotypic maintenance in postnatal life. Here, we report that the transcription factor NKX2.1 is present in mouse hypothalamic POMC neurons from early development to adulthood. Electromobility shift assays showed that NKX2.1 binds in vitro to NKX binding motifs present in the neuronal Pomc enhancers nPE1 and nPE2 and chromatin immunoprecipitation assays detected in vivo binding of NKX2.1 to nPE1 and nPE2 in mouse hypothalamic extracts. Transgenic and mutant studies performed in mouse embryos of either sex and adult males showed that the NKX motifs present in nPE1 and nPE2 are essential for their transcriptional enhancer activity. The conditional early inactivation of Nkx2.1 in the ventral hypothalamus prevented the onset of Pomc expression. Selective Nkx2.1 ablation from POMC neurons decreased Pomc expression in adult males and mildly increased their body weight and adiposity. Our results demonstrate that NKX2.1 is necessary to activate Pomc expression by binding to conserved canonical NKX motifs present in nPE1 and nPE2. Therefore, NKX2.1 plays a critical role in the early establishment of hypothalamic melanocortin neuron identity and participates in the maintenance of Pomc expression levels during adulthood. SIGNIFICANCE STATEMENT Food intake and body weight regulation depend on hypothalamic neurons that release satiety-inducing neuropeptides, known as melanocortins. Central melanocortins are encoded by Pomc , and Pomc mutations may lead to hyperphagia and severe obesity. Although the importance of central melanocortins is well appreciated, the genetic program that establishes and maintains fully functional POMC neurons remains to be explored. Here, we combined molecular, genetic, developmental, and functional studies that led to the discovery of NKX2.1, a transcription factor that participates in the early morphogenesis of the developing hypothalamus, as a key player in establishing the early identity of melanocortin neurons by activating Pomc expression. Thus, Nkx2.1 adds to the growing list of genes that participate in body weight regulation and adiposity., (Copyright © 2019 the authors.)
- Published
- 2019
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3. Selective Restoration of Pomc Expression in Glutamatergic POMC Neurons: Evidence for a Dynamic Hypothalamic Neurotransmitter Network.
- Author
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Jones GL, Wittmann G, Yokosawa EB, Yu H, Mercer AJ, Lechan RM, and Low MJ
- Subjects
- Animals, Female, Glutamic Acid metabolism, Male, Mice, Mice, Inbred C57BL, Obesity metabolism, Arcuate Nucleus of Hypothalamus metabolism, Energy Metabolism physiology, Neurons metabolism, Neurotransmitter Agents metabolism, Pro-Opiomelanocortin metabolism
- Abstract
Hypothalamic POMC deficiency leads to obesity and metabolic deficiencies, largely due to the loss of melanocortin peptides. However, POMC neurons in the arcuate nucleus (ARC) are comprised of glutamatergic and GABAergic subpopulations. The developmental program, relative proportion and function of these two subpopulations are unresolved. To test whether glutamatergic POMC neurons serve a distinct role in maintaining energy homeostasis, we activated Pomc expression Cre- dependently in Vglut2 -expressing neurons of mice with conditionally silenced Pomc alleles. The Vglut2 - Pomc restored mice had normal ARC Pomc mRNA levels, POMC immunoreactivity, as well as body weight and body composition at age 12 weeks. Unexpectedly, the cumulative total of Vglut2
+ glutamatergic- and Gad67+ GABAergic - Pomc neurons detected by in situ hybridization (ISH) exceeded 100% in both Vglut2 - Pomc restored and control mice, indicating that a subpopulation of Pomc neurons must express both neuronal markers. Consistent with this hypothesis, triple ISH of C57BL/6J hypothalami revealed that 35% of ARC Pomc neurons were selectively Gad67+ , 21% were selectively Vglut2+ , and 38% expressed both Gad67 and Vglut2 . The single Gad67+ and Vglut2+ Pomc neurons were most prevalent in the rostral ARC, while the Vglut2/Gad67+ dual-phenotype cells predominated in the caudal ARC. A lineage trace using Ai9-tdTomato reporter mice to label fluorescently all Vglut2 -expressing neurons showed equal numbers of tdTomato+ and tdTomato- POMC immunoreactive neurons. Together, these data suggest that POMC neurons exhibit developmental plasticity in their expression of glutamatergic and GABAergic markers, enabling re-establishment of normal energy homeostasis in the Vglut2 - Pomc restored mice.- Published
- 2019
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4. Central dopamine D2 receptors regulate growth-hormone-dependent body growth and pheromone signaling to conspecific males.
- Author
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Noaín D, Pérez-Millán MI, Bello EP, Luque GM, Casas Cordero R, Gelman DM, Peper M, Tornadu IG, Low MJ, Becú-Villalobos D, and Rubinstein M
- Subjects
- Analysis of Variance, Animals, Benzamides pharmacokinetics, Body Size drug effects, Body Size genetics, Body Weight drug effects, Body Weight genetics, Case-Control Studies, Catatonia chemically induced, Catatonia metabolism, Dopamine Antagonists pharmacology, Eating drug effects, Eating genetics, Eating physiology, Female, Haloperidol pharmacology, Insulin-Like Growth Factor I metabolism, Intermediate Filament Proteins genetics, Male, Mice, Mice, Inbred C57BL, Mice, Transgenic, Nerve Tissue Proteins genetics, Nestin, Oligodeoxyribonucleotides, Antisense pharmacology, Pheromones urine, Pituitary Gland drug effects, Prolactin genetics, Protein Binding drug effects, Protein Binding genetics, Proteins metabolism, Radioimmunoassay, Receptors, Dopamine D2 deficiency, Receptors, Dopamine D2 genetics, Social Dominance, Territoriality, Tritium pharmacokinetics, Body Size physiology, Growth Hormone metabolism, Pituitary Gland metabolism, Prolactin metabolism, Receptors, Dopamine D2 metabolism
- Abstract
Competition between adult males for limited resources such as food and receptive females is shaped by the male pattern of pituitary growth hormone (GH) secretion that determines body size and the production of urinary pheromones involved in male-to-male aggression. In the brain, dopamine (DA) provides incentive salience to stimuli that predict the availability of food and sexual partners. Although the importance of the GH axis and central DA neurotransmission in social dominance and fitness is clearly appreciated, the two systems have always been studied unconnectedly. Here we conducted a cell-specific genetic dissection study in conditional mutant mice that selectively lack DA D2 receptors (D2R) from pituitary lactotropes (lacDrd2KO) or neurons (neuroDrd2KO). Whereas lacDrd2KO mice developed a normal GH axis, neuroDrd2KO mice displayed fewer somatotropes; reduced hypothalamic Ghrh expression, pituitary GH content, and serum IGF-I levels; and exhibited reduced body size and weight. As a consequence of a GH axis deficit, neuroDrd2KO adult males excreted low levels of major urinary proteins and their urine failed to promote aggression and territorial behavior in control male challengers, in contrast to the urine taken from control adult males. These findings reveal that central D2Rs mediate a neuroendocrine-exocrine cascade that controls the maturation of the GH axis and downstream signals that are critical for fitness, social dominance, and competition between adult males.
- Published
- 2013
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5. Proopiomelanocortin expression in both GABA and glutamate neurons.
- Author
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Hentges ST, Otero-Corchon V, Pennock RL, King CM, and Low MJ
- Subjects
- Adrenocorticotropic Hormone metabolism, Animals, Cells, Cultured, Female, Glutamate Decarboxylase genetics, Glutamate Decarboxylase metabolism, Green Fluorescent Proteins genetics, Green Fluorescent Proteins metabolism, In Vitro Techniques, Luminescent Proteins genetics, Luminescent Proteins metabolism, Male, Mice, Mice, Inbred C57BL, Mice, Transgenic, Patch-Clamp Techniques, Pro-Opiomelanocortin genetics, Synapses physiology, Red Fluorescent Protein, Arcuate Nucleus of Hypothalamus physiology, Glutamic Acid metabolism, Neurons physiology, Pro-Opiomelanocortin metabolism, gamma-Aminobutyric Acid metabolism
- Abstract
Proopiomelanocortin (POMC) neurons have been intensively studied because of their essential role in regulating energy balance and body weight. Many effects of POMC neurons can be attributed to their release of cognate neuropeptides from secretory granules in axon terminals. However, these neurons also synaptically release non-peptide neurotransmitters. The aim of this study was to settle the controversy whether there are separate populations of POMC neurons that release GABA or glutamate. Transgenic mice expressing a red fluorescent protein [Discosoma red (DsRed)] driven by Pomc neuronal regulatory elements (POMC-DsRed) were crossed to mice that expressed green fluorescent protein (gfp) in GABAergic neurons (GAD67-gfp). Approximately 40% of POMC neurons in the arcuate nucleus of the double-transgenic mice expressed the GAD67-gfp transgene. In vitro neurotransmitter release was detected using whole-cell electrophysiologic recordings in cultured GAD67-gfp-positive and GAD67-gfp-negative POMC neurons that had formed recurrent synapses (autapses). Autapses from GAD67-gfp-positive neurons were uniformly GABAergic. In contrast, autapses from the GAD67-gfp-negative POMC neurons exclusively exhibited postsynaptic currents mediated by glutamate. Together, these results indicate that there are two subpopulations of POMC neurons in the arcuate nucleus differentiated by their amino acid neurotransmitter phenotype. Whole-cell voltage-clamp recordings from POMC neurons in live brain slices indicated that GABAergic and glutamatergic POMC neurons are under similar presynaptic and postsynaptic regulation, although the GABAergic POMC neurons are smaller and have higher input resistance. GABAergic and glutamatergic POMC neurons may mediate distinct aspects of POMC neuron function, including the regulation of energy homeostasis.
- Published
- 2009
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6. Visceral afferents directly activate catecholamine neurons in the solitary tract nucleus.
- Author
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Appleyard SM, Marks D, Kobayashi K, Okano H, Low MJ, and Andresen MC
- Subjects
- 4-Aminopyridine pharmacology, Analysis of Variance, Animals, Cholecystokinin pharmacology, Dose-Response Relationship, Radiation, Electric Stimulation methods, Excitatory Amino Acid Antagonists pharmacology, Green Fluorescent Proteins genetics, Green Fluorescent Proteins metabolism, In Vitro Techniques, Membrane Potentials radiation effects, Mice, Mice, Transgenic, Patch-Clamp Techniques methods, Potassium Channel Blockers pharmacology, Pyridazines pharmacology, Quinoxalines pharmacology, Tyrosine 3-Monooxygenase genetics, Tyrosine 3-Monooxygenase metabolism, Afferent Pathways physiology, Catecholamines metabolism, Membrane Potentials physiology, Neurons physiology, Solitary Nucleus cytology
- Abstract
Brainstem A2/C2 neurons are catecholamine (CA) neurons within the solitary tract nucleus (NTS) that influence many homeostatic functions, including cardiovascular reflexes, food intake, and stress. Because NTS is a major interface between sensory visceral afferents and the CNS, NTS CA neurons are ideally suited to coordinate complex responses by their projections to multiple brain regions. To test how NTS CA neurons process visceral afferent information carried by solitary tract (ST) afferents, we identified CA neurons using transgenic mice expressing TH-EGFP (enhanced green fluorescent protein under the control of the tyrosine hydroxylase promoter) and recorded synaptic responses to ST activation in horizontal slices. ST shocks evoked large-amplitude, short-latency, glutamatergic EPSCs (ST-EPSCs) in 90% of NTS CA neurons. Within neurons, ST-EPSCs had constant latency, rarely failed, and depressed substantially at high ST frequencies, indicating that NTS CA neurons receive direct monosynaptic connections from afferent terminals. NTS CA neurons received direct ST inputs from only one or two afferent fibers, with one-half also receiving smaller amplitude indirect inputs. Up to 90% of ST shocks evoked action potentials in NTS CA neurons. However, transmission of sensory afferent information through NTS CA neurons critically depended on the expression of an A-type potassium current (I(KA)), which when active attenuated ST-activated action potentials to a 37% success rate. The satiety peptide, cholecystokinin, presynaptically facilitated glutamate transmission in one-half of NTS CA neurons. Thus, NTS CA neurons are directly driven by visceral afferents with output being modulated by presynaptic peptide receptors and postsynaptic potassium channels.
- Published
- 2007
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7. Differential regulation of synaptic inputs by constitutively released endocannabinoids and exogenous cannabinoids.
- Author
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Hentges ST, Low MJ, and Williams JT
- Subjects
- Animals, Benzoxazines, Hypothalamus drug effects, Hypothalamus metabolism, Male, Mice, Mice, Inbred C57BL, Mice, Transgenic, Morpholines pharmacology, Naphthalenes pharmacology, Pro-Opiomelanocortin biosynthesis, Pro-Opiomelanocortin genetics, Receptor, Cannabinoid, CB1 antagonists & inhibitors, Receptor, Cannabinoid, CB1 metabolism, Cannabinoid Receptor Modulators metabolism, Cannabinoids pharmacology, Endocannabinoids, Synaptic Transmission drug effects, Synaptic Transmission physiology
- Abstract
Endocannabinoid release from a single neuron has been shown to cause presynaptic inhibition of transmitter release at many different sites. Here, we demonstrate that hypothalamic proopiomelanocortin (POMC) neurons release endocannabinoids continuously under basal conditions, unlike other release sites at which endocannabinoid production must be stimulated. The basal endocannabinoid release selectively inhibited GABA release onto POMC neurons, although exogenous administration of cannabinoid agonists also inhibited glutamate release. The CB1 cannabinoid receptor antagonist AM 251 [N-(piperidin-1-yl)-5-(4-iodophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide] blocked endocannabinoid-mediated inhibition of GABA release without affecting excitatory synaptic currents, whereas the CB1 receptor agonist WIN 55,212-2 [R-(+)-(2,3-dihydro-5-methyl-3-[(4-morpholinyl)methyl]pyrol [1,2,3-de]-1,4-benzoxazin-6-yl)(1-naphthalenyl) methanone monomethanesulfonate] inhibited both inhibitory and excitatory synaptic currents in POMC neurons. These data demonstrate that endogenously released cannabinoids and exogenously applied CB1 receptor agonists can have markedly different effects on synaptic inputs. Furthermore, the data suggest a novel form of endocannabinoid-mediated retrograde inhibition, whereby the regulation of a subset of inputs requires either the removal of tonic presynaptic inhibition caused by endocannabinoids or the engagement of a mechanism that actively inhibits endocannabinoid production.
- Published
- 2005
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8. Proopiomelanocortin neurons in nucleus tractus solitarius are activated by visceral afferents: regulation by cholecystokinin and opioids.
- Author
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Appleyard SM, Bailey TW, Doyle MW, Jin YH, Smart JL, Low MJ, and Andresen MC
- Subjects
- Animals, Cell Count methods, Dose-Response Relationship, Drug, Drug Interactions, Electric Stimulation methods, Enkephalin, Ala(2)-MePhe(4)-Gly(5)- pharmacology, Enkephalin, Methionine pharmacology, Excitatory Amino Acid Antagonists pharmacology, Excitatory Postsynaptic Potentials drug effects, Excitatory Postsynaptic Potentials physiology, Excitatory Postsynaptic Potentials radiation effects, Gene Expression Regulation drug effects, Green Fluorescent Proteins genetics, Hormone Antagonists pharmacology, Immunohistochemistry methods, In Vitro Techniques, Membrane Potentials drug effects, Membrane Potentials physiology, Membrane Potentials radiation effects, Mice, Mice, Transgenic, Neurons metabolism, Patch-Clamp Techniques methods, Pro-Opiomelanocortin genetics, Proglumide analogs & derivatives, Proglumide pharmacology, Proto-Oncogene Proteins c-fos metabolism, Quinoxalines pharmacology, Time Factors, Cholecystokinin pharmacology, Narcotics pharmacology, Neurons drug effects, Pro-Opiomelanocortin metabolism, Solitary Nucleus cytology, Visceral Afferents physiology
- Abstract
The nucleus tractus solitarius (NTS) receives dense terminations from cranial visceral afferents, including those from the gastrointestinal (GI) system. Although the NTS integrates peripheral satiety signals and relays this signal to central feeding centers, little is known about which NTS neurons are involved or what mechanisms are responsible. Proopiomelanocortin (POMC) neurons are good candidates for GI integration, because disruption of the POMC gene leads to severe obesity and hyperphagia. Here, we used POMC-enhanced green fluorescent protein (EGFP) transgenic mice to identify NTS POMC neurons. Intraperitoneal administration of cholecystokinin (CCK) induced c-fos gene expression in NTS POMC-EGFP neurons, suggesting that they are activated by afferents stimulated by the satiety hormone. We tested the synaptic relationship of these neurons to visceral afferents and their modulation by CCK and opioids using patch recordings in horizontal brain slices. Electrical activation of the solitary tract (ST) evoked EPSCs in NTS POMC-EGFP neurons. The invariant latencies, low failure rates, and substantial paired-pulse depression of the ST-evoked EPSCs indicate that NTS POMC-EGFP neurons are second-order neurons directly contacted by afferent terminals. The EPSCs were blocked by the glutamate antagonist 2,3-dihydroxy-6-nitro-7-sulfonyl-benzo[f]quinoxaline. CCK increased the amplitude of the ST-stimulated EPSCs and the frequency of miniature EPSCs, effects attenuated by the CCK1 receptor antagonist lorglumide. In contrast, the orexigenic opioid agonists [D-Ala(2), N-Me-Phe(4), Gly-ol(5)]-enkephalin and met-enkephalin inhibited both ST-stimulated EPSCs and the frequency of miniature EPSCs. These findings identify a potential satiety pathway in which visceral afferents directly activate NTS POMC-EGFP neurons with excitatory inputs that are appropriately modulated by appetite regulators.
- Published
- 2005
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9. A transgenic marker for newly born granule cells in dentate gyrus.
- Author
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Overstreet LS, Hentges ST, Bumaschny VF, de Souza FS, Smart JL, Santangelo AM, Low MJ, Westbrook GL, and Rubinstein M
- Subjects
- Action Potentials physiology, Aging metabolism, Animals, Biomarkers analysis, Bromodeoxyuridine, Cell Count, Cell Division physiology, Cell Movement, Dentate Gyrus cytology, Genes, Reporter, Green Fluorescent Proteins, Luminescent Proteins biosynthesis, Luminescent Proteins genetics, Mice, Mice, Inbred C57BL, Mice, Transgenic, Neural Cell Adhesion Molecule L1 biosynthesis, Physical Exertion physiology, Pro-Opiomelanocortin genetics, Promoter Regions, Genetic physiology, Sialic Acids biosynthesis, Dentate Gyrus metabolism, Neurons metabolism, Transgenes physiology
- Abstract
Neurogenesis in the dentate gyrus continues into adulthood, yet little is known about the function of newly born neurons or how they integrate into an existing network of mature neurons. We made transgenic mice that selectively and transiently express enhanced green fluorescent protein (EGFP) in newly born granule cells of the dentate gyrus under the transcriptional control of proopiomelanocortin (POMC) genomic sequences. Analysis of transgenic pedigrees with truncation or deletion mutations indicated that EGFP expression in the dentate gyrus required cryptic POMC promoter regions dispensable for arcuate hypothalamic or pituitary expression. Unlike arcuate neurons, dentate granule cells did not express the endogenous POMC gene. EGFP-positive neurons had immature properties, including short spineless dendrites and small action potentials. Colocalization with bromodeoxyuridine indicated that EGFP-labeled granule cells were approximately 2 weeks postmitotic. EGFP-labeled cells expressed markers for immature granule cells but not the glial marker GFAP. The number of EGFP-labeled neurons declined with age and increased with exercise, paralleling neurogenesis. Our results indicate that POMC-EGFP marks immature granule cells and that adult-generated granule cells integrate quite slowly into the hippocampal circuitry.
- Published
- 2004
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10. GABA release from proopiomelanocortin neurons.
- Author
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Hentges ST, Nishiyama M, Overstreet LS, Stenzel-Poore M, Williams JT, and Low MJ
- Subjects
- Animals, Arcuate Nucleus of Hypothalamus, Carrier Proteins metabolism, Cells, Cultured, GABA Plasma Membrane Transport Proteins, Green Fluorescent Proteins, Hypothalamus cytology, Luminescent Proteins genetics, Membrane Proteins metabolism, Mice, Mice, Inbred C57BL, Mice, Transgenic, Narcotics pharmacology, Neurons cytology, Neurons drug effects, Patch-Clamp Techniques, Presynaptic Terminals metabolism, Pro-Opiomelanocortin genetics, Recombinant Fusion Proteins genetics, Recombinant Fusion Proteins metabolism, Membrane Transport Proteins, Neurons metabolism, Organic Anion Transporters, Pro-Opiomelanocortin metabolism, gamma-Aminobutyric Acid metabolism
- Abstract
Neural networks controlling food intake and energy homeostasis clearly involve proopiomelanocortin (POMC) neurons and their peptide transmitters. alpha-melanocyte-stimulating hormone from arcuate POMC neurons potently reduces food intake, whereas arcuate neuropeptide Y (NPY) neurons act in opposition to stimulate food intake. In addition to orexigenic peptides, NPY neurons also release the inhibitory neurotransmitter GABA, which can act in a local circuit to inhibit POMC neuron activity. Whether or not reciprocal inhibition could occur has not yet been determined, because the presence of a rapid neurotransmitter in POMC neurons has not been demonstrated previously. Here, we used primary cultures of fluorescently labeled POMC neurons that had formed recurrent synapses (autapses) to detect the release of neurotransmitter. When an action potential was evoked in the axon of a POMC neuron with autapses, a short-latency synaptic current was recorded in the same cell. The autaptic current was abolished by GABA(A) receptor antagonists and substantially inhibited by opioids. Double-label in situ RNA hybridization for POMC and glutamic acid decarboxylase, the GABA synthetic enzyme, revealed colocalization of mRNAs in approximately one-third of POMC neurons in vivo. Our results suggest that these neurons can exert rapid inhibitory effects via the release of GABA, in addition to the more sustained actions provided by POMC peptides. However, this rapid inhibition may not play a major role within local hypothalamic circuits, but rather is likely to be important in more distant projection areas as indicated by the colocalization of vesicular GABA transporter immunoreactivity predominantly in extrahypothalamic POMC terminals.
- Published
- 2004
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11. Differential effects of direct and indirect dopamine agonists on prepulse inhibition: a study in D1 and D2 receptor knock-out mice.
- Author
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Ralph-Williams RJ, Lehmann-Masten V, Otero-Corchon V, Low MJ, and Geyer MA
- Subjects
- Amphetamine pharmacology, Animals, Apomorphine pharmacology, Behavior, Animal drug effects, Behavior, Animal physiology, Benzazepines pharmacology, Dizocilpine Maleate pharmacology, Electric Stimulation, Excitatory Amino Acid Antagonists pharmacology, Female, Genotype, Heterozygote, Homozygote, Male, Mice, Mice, Inbred C57BL, Mice, Knockout, Neural Inhibition physiology, Receptors, Dopamine D1 genetics, Receptors, Dopamine D2 genetics, Receptors, N-Methyl-D-Aspartate antagonists & inhibitors, Reflex, Startle drug effects, Reflex, Startle genetics, Dopamine Agonists pharmacology, Neural Inhibition drug effects, Receptors, Dopamine D1 agonists, Receptors, Dopamine D1 deficiency, Receptors, Dopamine D2 agonists, Receptors, Dopamine D2 deficiency
- Abstract
Stimulation of the dopamine (DA) system disrupts prepulse inhibition (PPI) of the acoustic startle response. On the basis of rat studies, it appeared that DA D2 receptors (D2Rs) rather than D1 receptors (D1Rs) regulate PPI, albeit possibly in synergism with D1Rs. To characterize the DA receptor modulation of PPI in another species, we tested DA D1R and D2R mutant mice with direct and indirect DA agonists and with the glutamate receptor antagonist, dizocilpine (MK-801). Neither the mixed D1/D2 agonist apomorphine (5 mg/kg) nor the more selective D1-like agonist SKF82958 (0.3 mg/kg) altered PPI in D1R knock-out mice, although both compounds disrupted PPI in D2R mutant and wild-type mice, suggesting that the D1R alone might modulate PPI in mice. However, amphetamine (10 mg/kg) significantly lowered PPI in each genotype of D1R mice, suggesting that the D1R is not necessary for the PPI-disruptive effect of the indirect agonist in mice. As reported previously, amphetamine (10 mg/kg) failed to disrupt PPI in D2R knock-out mice, supporting a unique role of the D2R in the modulation of PPI. Dizocilpine (0.3 mg/kg) induced similar PPI deficits in D1R and D2R mutant mice, confirming that the influences of the NMDA receptor on PPI are independent of D1Rs and D2Rs in rodents. Thus, both D1Rs and D2Rs modulate aspects of PPI in mice in a manner that differs from dopaminergic modulation in rats. These findings emphasize that further cross-species comparisons of the pharmacology of PPI are essential to understand the relevance of rodent PPI studies to the deficits in PPI observed in patients with schizophrenia.
- Published
- 2002
12. Selective reward deficit in mice lacking beta-endorphin and enkephalin.
- Author
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Hayward MD, Pintar JE, and Low MJ
- Subjects
- Animals, Choice Behavior, Conditioning, Operant physiology, Eating physiology, Enkephalins genetics, Extinction, Psychological physiology, Feeding Behavior, Food Deprivation, Genotype, Male, Mice, Mice, Knockout, Motivation, Phenotype, beta-Endorphin genetics, Behavior, Animal physiology, Enkephalins deficiency, Reward, beta-Endorphin deficiency
- Abstract
It has been impossible to unequivocally identify which endogenous opioids modulate the incentive value of rewarding stimuli because these peptides are not highly selective for any single opioid receptor subtype. Here, we present evidence based on the measurement of instrumental behavior of beta-endorphin and enkephalin knock-out mice that both opioid peptides play a positive role. A progressive ratio schedule was used to measure how hard an animal would work for food reinforcers. The loss of either opioid reduced responding under this schedule, regardless of the palatability of the three different formulas of reinforcers used. The phenotype of mice lacking both endogenous opioids was nearly identical to the phenotype of mice mutant for either individual opioid. Responses were tested in nondeprived and deprived feeding states but were reduced in beta-endorphin- and enkephalin-deficient mice only when they were maintained under nondeprived conditions. Other operant manipulations ruled out variables that might contribute nonspecifically to this result such as differences in acquisition, early satiation, motor performance deficit, and reduced resistance to extinction. In contrast to the effects on instrumental performance, the loss of either or both endogenous opioids did not influence preference for water flavored with sucrose or saccharin in a two-bottle free-choice drinking paradigm. We conclude that both beta-endorphin and enkephalin positively contribute to the incentive-motivation to acquire food reinforcers. Because the attenuation of operant responding was observed only during a nondeprived motivational state, the hedonics of feeding are likely altered rather than energy homeostasis.
- Published
- 2002
13. Failure of intravenous morphine to serve as an effective instrumental reinforcer in dopamine D2 receptor knock-out mice.
- Author
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Elmer GI, Pieper JO, Rubinstein M, Low MJ, Grandy DK, and Wise RA
- Subjects
- Animals, Behavior, Animal drug effects, Conditioning, Operant drug effects, Dose-Response Relationship, Drug, Drinking genetics, Heterozygote, Homozygote, Injections, Intravenous, Male, Mice, Mice, Knockout, Receptors, Dopamine D2 genetics, Reward, Self Administration, Morphine administration & dosage, Receptors, Dopamine D2 deficiency, Reinforcement, Psychology
- Abstract
The rewarding effects of opiates are thought to be mediated through dopaminergic mechanisms in the ventral tegmental area, dopamine-independent mechanisms in the nucleus accumbens, or both. The purpose of the present study was to explore the contribution of dopamine to opiate-reinforced behavior using D2 receptor knock-out mice. Wild-type, heterozygous, and D2 knock-out mice were first trained to lever press for water reinforcement and then implanted with intravenous catheters. The ability of intravenously delivered morphine to maintain lever pressing in these mice was studied under two schedules of reinforcement: a fixed ratio 4 (FR4) schedule (saline, 0.1, 0.3, or 1.0 mg/kg, per injection) and a progressive ratio (PR) schedule (1.0 mg/kg, per injection). In the wild-type and heterozygous mice, FR4 behavior maintained by morphine injections was significantly greater than behavior maintained by vehicle injections. Response rate was inversely related to injection dose and increased significantly in the wild-type and heterozygous mice when the animals were placed on the PR schedule. In contrast, the knock-out mice did not respond more for morphine than for saline and did not respond more when increased ratios were required by the PR schedule. Thus, morphine served as a positive reinforcer in the wild-type and heterozygous mice but failed to do so in the knock-out mice. Under this range of doses and response requirements, the rewarding effects of morphine appear to depend critically on an intact D2 receptor system.
- Published
- 2002
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14. Dysfunctional light-evoked regulation of cAMP in photoreceptors and abnormal retinal adaptation in mice lacking dopamine D4 receptors.
- Author
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Nir I, Harrison JM, Haque R, Low MJ, Grandy DK, Rubinstein M, and Iuvone PM
- Subjects
- Animals, Cell Survival genetics, Darkness, Dopamine metabolism, Dopamine Agonists pharmacology, Glutamic Acid metabolism, In Vitro Techniques, Light, Mice, Mice, Inbred C57BL, Mice, Knockout, Phagocytosis physiology, Photoreceptor Cells, Vertebrate drug effects, Photoreceptor Cells, Vertebrate radiation effects, Quinpirole pharmacology, Receptors, Dopamine D2 agonists, Receptors, Dopamine D2 genetics, Receptors, Dopamine D4, Retina drug effects, Retina metabolism, Retina radiation effects, Rod Cell Outer Segment ultrastructure, Adaptation, Ocular physiology, Cyclic AMP metabolism, Photoreceptor Cells, Vertebrate metabolism, Receptors, Dopamine D2 deficiency
- Abstract
Dopamine is a retinal neuromodulator that has been implicated in many aspects of retinal physiology. Photoreceptor cells express dopamine D4 receptors that regulate cAMP metabolism. To assess the effects of dopamine on photoreceptor physiology, we examined the morphology, electrophysiology, and regulation of cAMP metabolism in mice with targeted disruption of the dopamine D4 receptor gene. Photoreceptor morphology and outer segment disc shedding after light onset were normal in D4 knock-out (D4KO) mice. Quinpirole, a dopamine D2/D3/D4 receptor agonist, decreased cAMP synthesis in retinas of wild-type (WT) mice but not in retinas of D4KO mice. In WT retinas, the photoreceptors of which were functionally isolated by incubation in the presence of exogenous glutamate, light also suppressed cAMP synthesis. Despite the similar inhibition of cAMP synthesis, the effect of light is directly on the photoreceptors and independent of dopamine modulation, because it was unaffected by application of the D4 receptor antagonist l-745,870. Nevertheless, compared with WT retinas, basal cAMP formation was reduced in the photoreceptors of D4KO retinas, and light had no additional inhibitory effect. The results suggest that dopamine, via D4 receptors, normally modulates the cascade that couples light responses to adenylyl cyclase activity in photoreceptor cells, and the absence of this modulation results in dysfunction of the cascade. Dark-adapted electroretinogram (ERG) responses were normal in D4KO mice. However, ERG b-wave responses were greatly suppressed during both light adaptation and early stages of dark adaptation. Thus, the absence of D4 receptors affects adaptation, altering transmission of light responses from photoreceptors to inner retinal neurons. These findings indicate that dopamine D4 receptors normally play a major role in regulating photoreceptor cAMP metabolism and adaptive retinal responses to changing environmental illumination.
- Published
- 2002
15. Dopamine D4 receptor-deficient mice display cortical hyperexcitability.
- Author
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Rubinstein M, Cepeda C, Hurst RS, Flores-Hernandez J, Ariano MA, Falzone TL, Kozell LB, Meshul CK, Bunzow JR, Low MJ, Levine MS, and Grandy DK
- Subjects
- 4-Aminopyridine pharmacology, Animals, Bicuculline pharmacology, Cerebral Cortex drug effects, Convulsants pharmacology, Dopamine metabolism, Dopamine D2 Receptor Antagonists, Dose-Response Relationship, Drug, Glutamic Acid metabolism, Immunohistochemistry, In Vitro Techniques, Membrane Potentials drug effects, Mice, Mice, Neurologic Mutants, Motor Cortex drug effects, Motor Cortex metabolism, Motor Cortex physiopathology, Neural Inhibition drug effects, Neural Inhibition genetics, Patch-Clamp Techniques, Piperazines pharmacology, Presynaptic Terminals metabolism, Pyramidal Cells drug effects, Pyramidal Cells metabolism, Receptors, Dopamine D2 genetics, Receptors, Dopamine D4, Seizures chemically induced, Sulfonamides pharmacology, Cerebral Cortex physiopathology, Receptors, Dopamine D2 deficiency, Seizures physiopathology
- Abstract
The dopamine D(4) receptor (D(4)R) is predominantly expressed in the frontal cortex (FC), a brain region that receives dense input from midbrain dopamine (DA) neurons and is associated with cognitive and emotional processes. However, the physiological significance of this dopamine receptor subtype has been difficult to explore because of the slow development of D(4)R agonists and antagonists the selectivity and efficacy of which have been rigorously demonstrated in vivo. We have attempted to overcome this limitation by taking a multidimensional approach to the characterization of mice completely deficient in this receptor subtype. Electrophysiological current and voltage-clamp recordings were performed in cortical pyramidal neurons from wild-type and D(4)R-deficient mice. The frequency of spontaneous synaptic activity and the frequency and duration of paroxysmal discharges induced by epileptogenic agents were increased in mutant mice. Enhanced synaptic activity was also observed in brain slices of wild-type mice incubated in the presence of the selective D(4)R antagonist PNU-101387G. Consistent with greater electrophysiological activity, nerve terminal glutamate density associated with asymmetrical synaptic contacts within layer VI of the motor cortex was reduced in mutant neurons. Taken together, these results suggest that the D(4)R can function as an inhibitory modulator of glutamate activity in the FC.
- Published
- 2001
16. Functional uncoupling of adenosine A(2A) receptors and reduced responseto caffeine in mice lacking dopamine D2 receptors.
- Author
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Zahniser NR, Simosky JK, Mayfield RD, Negri CA, Hanania T, Larson GA, Kelly MA, Grandy DK, Rubinstein M, Low MJ, and Fredholm BB
- Subjects
- Animals, Basal Ganglia Diseases drug therapy, Basal Ganglia Diseases physiopathology, Cyclic AMP metabolism, Female, Globus Pallidus cytology, Globus Pallidus metabolism, In Vitro Techniques, Male, Mice, Mice, Congenic, Mice, Knockout, Motor Activity drug effects, Motor Activity physiology, Neostriatum cytology, Neostriatum metabolism, Neural Pathways cytology, Neural Pathways metabolism, Proto-Oncogene Proteins c-fos drug effects, Proto-Oncogene Proteins c-fos metabolism, RNA, Messenger drug effects, RNA, Messenger metabolism, Receptor, Adenosine A2A, Receptors, Dopamine D2 drug effects, Receptors, Dopamine D2 genetics, Receptors, Purinergic P1 drug effects, Receptors, Purinergic P1 genetics, Caffeine pharmacology, Globus Pallidus drug effects, Neostriatum drug effects, Neural Pathways drug effects, Receptors, Dopamine D2 metabolism, Receptors, Purinergic P1 metabolism, gamma-Aminobutyric Acid metabolism
- Abstract
Dopamine D(2) receptors (Rs) and adenosine A(2A)Rs are coexpressed on striatopallidal neurons, where they mediate opposing actions. In agreement with the idea that D(2)Rs tonically inhibit GABA release from these neurons, stimulation-evoked GABA release was significantly greater from striatal/pallidal slices from D(2)R null mutant (D(2)R(-/-)) than from wild-type (D(2)R(+/+)) mice. Release from heterozygous (D(2)R(+/-)) slices was intermediate. However, contrary to predictions that A(2A)R effects would be enhanced in D(2)R-deficient mice, the A(2A)R agonist CGS 21680 significantly increased GABA release only from D(2)R(+/+) slices. CGS 21680 modulation was observed when D(2)Rs were antagonized by raclopride, suggesting that an acute absence of D(2)Rs cannot explain the results. The lack of CGS 21680 modulation in the D(2)R-deficient mice was also not caused by a compensatory downregulation of A(2A)Rs in the striatum or globus pallidus. However, CGS 21680 significantly stimulated cAMP production only in D(2)R(+/+) striatal/pallidal slices. This functional uncoupling of A(2A)Rs in the D(2)R-deficient mice was not explained by reduced expression of G(s), G(olf), or type VI adenylyl cyclase. Locomotor activity induced by the adenosine receptor antagonist caffeine was significantly less pronounced in D(2)R(-/-) mice than in D(2)R(+/+) and D(2)R(+/-) mice, further supporting the idea that D(2)Rs are required for caffeine activation. Caffeine increased c-fos only in D(2)R(-/-) globus pallidus. The present results show that a targeted disruption of the D(2)R reduces coupling of A(2A)Rs on striatopallidal neurons and thereby responses to drugs that act on adenosine receptors. They also reinforce the ideas that D(2)Rs and A(2A)Rs are functionally opposed and that D(2)R-mediated effects normally predominate.
- Published
- 2000
17. Dopamine D4 receptor-knock-out mice exhibit reduced exploration of novel stimuli.
- Author
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Dulawa SC, Grandy DK, Low MJ, Paulus MP, and Geyer MA
- Subjects
- Analysis of Variance, Animals, Avoidance Learning physiology, Female, Humans, Male, Mice, Mice, Knockout, Motor Activity physiology, Polymorphism, Genetic, Reaction Time, Receptors, Dopamine D2 deficiency, Receptors, Dopamine D2 genetics, Receptors, Dopamine D4, Exploratory Behavior physiology, Receptors, Dopamine D2 physiology
- Abstract
The involvement of dopamine neurotransmission in behavioral responses to novelty is suggested by reports that reward is related to increased dopamine activity, that dopamine modulates exploratory behavior in animals, and that Parkinson's disease patients report diminished responses to novelty. Some studies have reported that polymorphisms of the human dopamine D4 receptor (D4R) gene are associated with personality inventory measures of the trait called "novelty-seeking". To explore a potential role for the D4R in behavioral responses to novelty, we evaluated D4R-knock-out (D4R-/-) and wild-type (D4R+/+) mice in three approach-avoidance paradigms: the open field, emergence, and novel object tests. These three paradigms differ in the degree to which they elicit approach, or exploratory behavior, and avoidance, or anxiety-related behavior. Thus, we used these three tests to determine whether the D4R primarily influences the exploratory or the anxious component of responses to approach-avoidance conflicts. D4R-/- mice were significantly less behaviorally responsive to novelty than D4R+/+ mice in all three tests. The largest phenotypic differences were observed in the novel object test, which maximizes approach behavior, and the smallest phenotypic differences were found in the open field test, which maximizes avoidance behavior. Hence, D4R-/- mice exhibit reductions in behavioral responses to novelty, reflecting a decrease in novelty-related exploration.
- Published
- 1999
18. The dopamine D2, but not D3 or D4, receptor subtype is essential for the disruption of prepulse inhibition produced by amphetamine in mice.
- Author
-
Ralph RJ, Varty GB, Kelly MA, Wang YM, Caron MG, Rubinstein M, Grandy DK, Low MJ, and Geyer MA
- Subjects
- Acoustic Stimulation, Animals, Mice, Mice, Knockout, Mice, Mutant Strains, Receptors, Dopamine D2 drug effects, Receptors, Dopamine D3, Receptors, Dopamine D4, Reflex, Startle drug effects, Amphetamine pharmacology, Dopamine Agents pharmacology, Neural Inhibition drug effects, Receptors, Dopamine drug effects
- Abstract
Brain dopamine (DA) systems are involved in the modulation of the sensorimotor gating phenomenon known as prepulse inhibition (PPI). The class of D2-like receptors, including the D2, D3, and D4 receptor subtypes, have all been implicated in the control of PPI via studies of DA agonists and antagonists in rats. Nevertheless, the functional relevance of each receptor subtype remains unclear because these ligands are not specific. To determine the relevance of each receptor subtype, we used genetically altered strains of "knock-out" mice lacking the DA D2, D3, or D4 receptors. We tested the effects of each knock-out on both the phenotypic expression of PPI and the disruption of PPI produced by the indirect DA agonist d-amphetamine (AMPH). No phenotypic differences in PPI were observed at baseline. AMPH significantly disrupted PPI in the D2 (+/+) mice but had no effect in the D2 (-/-) mice. After AMPH treatment, both DA D3 and D4 receptor (+/+) and (-/-) mice had significant disruptions in PPI. These findings indicate that the AMPH-induced disruption of PPI is mediated via the DA D2 receptor and not the D3 or D4 receptor subtypes. Uncovering the neural mechanisms involved in PPI will further our understanding of the substrates of sensorimotor gating and could lead to better therapeutics to treat gating disorders, such as schizophrenia.
- Published
- 1999
19. Authentic cell-specific and developmentally regulated expression of pro-opiomelanocortin genomic fragments in hypothalamic and hindbrain neurons of transgenic mice.
- Author
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Young JI, Otero V, Cerdán MG, Falzone TL, Chan EC, Low MJ, and Rubinstein M
- Subjects
- Animals, Hypothalamus cytology, Mice, Mice, Transgenic, Neurons metabolism, Organ Specificity, Rhombencephalon cytology, DNA Fragmentation, Gene Expression Regulation, Developmental physiology, Genome, Hypothalamus metabolism, Pro-Opiomelanocortin genetics, Rhombencephalon metabolism
- Abstract
The pro-opiomelanocortin (POMC) gene is expressed in a subset of hypothalamic and hindbrain neurons and in pituitary melanotrophs and corticotrophs. POMC neurons release the potent opioid beta-endorphin and several active melanocortins that control homeostasis and feeding behavior. POMC gene expression in the CNS is believed to be controlled by distinct cis-acting regulatory sequences. To analyze the transcriptional regulation of POMC in neuronal and endocrine cells, we produced transgenic mice carrying POMC27*, a transgene containing the entire 6 kb of the POMC transcriptional unit together with 13 kb of 5' flanking regions and 8 kb of 3' flanking regions. POMC27* was tagged with a heterologous 30 bp oligonucleotide in the third exon. In situ hybridization studies showed an accurate cell-specific pattern of expression of POMC27* in the arcuate nucleus and the pituitary. Hypothalamic mRNA-positive neurons colocalized entirely with beta-endorphin immunoreactivity. No ectopic transgenic expression was detected in the brain. Deletional analyses demonstrated that neuron-specific expression of POMC transgenes required distal 5' sequences localized upstream of the pituitary-responsive proximal cis-acting elements that were identified previously. POMC27* exhibited a spatial and temporal pattern of expression throughout development that exactly paralleled endogenous POMC. RNase protection assays revealed that POMC27* expression mimicked that of POMC in different areas of the CNS and most peripheral organs with no detectable ectopic expression. Hormonal regulation of POMC27* and POMC was identical in the hypothalamus and pituitary. These results show that distal 5' sequences of the POMC gene located between -13 and -2 kb target expression into the CNS of transgenic mice in a precise neuron-specific, developmentally and hormonally regulated manner.
- Published
- 1998
20. Locomotor activity in D2 dopamine receptor-deficient mice is determined by gene dosage, genetic background, and developmental adaptations.
- Author
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Kelly MA, Rubinstein M, Phillips TJ, Lessov CN, Burkhart-Kasch S, Zhang G, Bunzow JR, Fang Y, Gerhardt GA, Grandy DK, and Low MJ
- Subjects
- Animals, Corpus Striatum physiology, Dopamine Antagonists pharmacology, Dopamine D2 Receptor Antagonists, Female, Haloperidol pharmacology, Male, Methyltyrosines pharmacology, Mice, Mice, Inbred C57BL, Movement physiology, Neural Pathways physiology, Psychomotor Performance physiology, Reserpine pharmacology, Substantia Nigra physiology, Adaptation, Physiological, Gene Dosage, Motor Activity genetics, Receptors, Dopamine D2 deficiency
- Abstract
Locomotor activity is a polygenic trait that varies widely among inbred strains of mice (). To characterize the role of D2 dopamine receptors in locomotion, we generated F2 hybrid (129/Sv x C57BL/6) D2 dopamine receptor (D2R)-deficient mice by gene targeting and investigated the contribution of genetic background to open-field activity and rotarod performance. Horizontal activity of D2R-/- mice was approximately half that of drug-naive, strain-matched controls but was significantly greater than haloperidol-treated controls, which were markedly hypokinetic. Wild-type 129/SvEv and C57BL/6 mice with functional D2 receptors had greater interstrain differences in spontaneous activity than those among the F2 hybrid mutants. Incipient congenic strains of D2R-deficient mice demonstrated an orderly gene dosage reduction in locomotion superimposed on both extremes of parental background locomotor activity. In contrast, F2 hybrid D2R-/- mice had impaired motor coordination on the rotarod that was corrected in the congenic C57BL/6 background. Wild-type 129/SvEv mice had the poorest rotarod ability of all groups tested, suggesting that linked substrain 129 alleles, not the absence of D2 receptors per se, were largely responsible for the reduced function of the F2 hybrid D2R-/- and D2R+/- mice. Neurochemical and pharmacological studies revealed unexpectedly normal tissue striatal monoamine levels and no evidence for supersensitive D1, D3, or D4 dopamine receptors in the D2R-/- mice. However, after acute monoamine depletion, akinetic D2R+/- mice had a significantly greater synergistic restoration of locomotion in response to SKF38393 and quinpirole compared with any group of D2R+/+ controls. We conclude that D2R-deficient mice are not a model of Parkinson's disease. Our studies highlight the interaction of multiple genetic factors in the analysis of complex behaviors in gene knock-out mice.
- Published
- 1998
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