38 results on '"Herkenham M"'
Search Results
2. Environmental enrichment requires adult neurogenesis to facilitate the recovery from psychosocial stress
- Author
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Schloesser, R J, Lehmann, M, Martinowich, K, Manji, H K, and Herkenham, M
- Published
- 2010
- Full Text
- View/download PDF
3. Lymphocytes from Chronically Stressed Mice Confer Antidepressant-Like Effects to Naive Mice
- Author
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Brachman, R. A., primary, Lehmann, M. L., additional, Maric, D., additional, and Herkenham, M., additional
- Published
- 2015
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4. Glucocorticoids Orchestrate Divergent Effects on Mood through Adult Neurogenesis
- Author
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Lehmann, M. L., primary, Brachman, R. A., additional, Martinowich, K., additional, Schloesser, R. J., additional, and Herkenham, M., additional
- Published
- 2013
- Full Text
- View/download PDF
5. Environmental Enrichment Confers Stress Resiliency to Social Defeat through an Infralimbic Cortex-Dependent Neuroanatomical Pathway
- Author
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Lehmann, M. L., primary and Herkenham, M., additional
- Published
- 2011
- Full Text
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6. Repeated electroconvulsive shock produces long-lasting increases in messenger RNA expression of corticotropin-releasing hormone and tyrosine hydroxylase in rat brain. Therapeutic implications.
- Author
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Brady, L S, primary, Lynn, A B, additional, Glowa, J R, additional, Le, D Q, additional, and Herkenham, M, additional
- Published
- 1994
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7. Systemic interleukin-1 induces early and late patterns of c-fos mRNA expression in brain
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Brady, LS, primary, Lynn, AB, additional, Herkenham, M, additional, and Gottesfeld, Z, additional
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- 1994
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8. Long-term antidepressant administration alters corticotropin-releasing hormone, tyrosine hydroxylase, and mineralocorticoid receptor gene expression in rat brain. Therapeutic implications.
- Author
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Brady, L S, primary, Whitfield, H J, additional, Fox, R J, additional, Gold, P W, additional, and Herkenham, M, additional
- Published
- 1991
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9. Characterization and localization of cannabinoid receptors in rat brain: a quantitative in vitro autoradiographic study
- Author
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Herkenham, M, primary, Lynn, AB, additional, Johnson, MR, additional, Melvin, LS, additional, de Costa, BR, additional, and Rice, KC, additional
- Published
- 1991
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10. Cannabinoid receptor localization in brain.
- Author
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Herkenham, M, primary, Lynn, A B, additional, Little, M D, additional, Johnson, M R, additional, Melvin, L S, additional, de Costa, B R, additional, and Rice, K C, additional
- Published
- 1990
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11. Intraneuronal generation of a pyridinium metabolite may cause drug-induced parkinsonism.
- Author
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Markey, S. P., Johannessen, J. N., Chiueh, C. C., Burns, R. S., and Herkenham, M. A.
- Published
- 1984
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12. In vitro autoradiography of opiate receptors in rat brain suggests loci of "opiatergic" pathways.
- Author
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Herkenham, M and Pert, C B
- Abstract
Slide-mounted sections of unfixed frozen rat brain can be labeled in vitro with [3H]naloxone to show the mu-like ligand selectivity characterized in previous studies. We have developed an autoradiographic technique using hot paraformaldehyde vapors to prevent diffusion of ligands with reversible binding. Resolution at the light level is sufficient to detect concordance between receptor patterns and terminal fields of axonal projections marked by tract-tracing techniques. The opiate receptor distribution suggests the existence of widespread intrinsic and several longer multisynaptic "opiatergic pathways within sensory and limbic circuits. One multisynaptic pathway may link olfactory structures with limbic circuits in the amygdala and habenula. Another may lie in limbic cortical structures. Opiate receptors are numerous also in sensory systems, and within primary sensory nuclei (visual, auditory, olfactory, somatic) they are found superficially in laminated structures. Together, the opiate receptors are well placed to control incoming sensory and subsequent limbic information processing.
- Published
- 1980
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13. Interconverting mu and delta forms of the opiate receptor in rat striatal patches.
- Author
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Bowen, W D, Gentleman, S, Herkenham, M, and Pert, C B
- Abstract
The binding of a radiolabeled "mu receptor" prototype opiate, dihydromorphine (H2morphine), and the binding of a "delta receptor" prototype, [D-Ala2,D-Leu5]enkephalin (D-Enk), to slide-mounted rat caudate slices were simultaneously compared quantitatively and visualized by autoradiography. Generally, D-Enk bound to opiate receptors distributed evenly throughout the entire striatum (type 2 pattern), whereas H2morphine labeled discrete islands or patches of receptors (type 1 pattern). In the presence of Mn2+ (3 mM) or other divalent cations, however, Na+ and GTP at 25 degrees C caused an increase in D-Enk binding at the expense of H2morphine binding at striatal opiate receptor patches. Thus, these conditions shifted D-Enk binding from an even pattern to one that included both an even and patchy distribution. These incubation conditions not only promoted D-Enk binding to striatal patches but also enabled the opiate receptor to regulate adenylate cyclase with the same (P less than 0.01) ligand selectivity pattern as that obtained by the displacement of D-Enk binding. The relative affinity of opiate receptors in striatal patches for opiate peptides, naloxone, and morphine appears to be a function of incubation conditions and coupling to adenylate cyclase and is not indicative of distinctly different opiate receptors. We postulate a three-state allosteric model consisting of mu agonist-, mu antagonists-, and adenylate cyclase-coupled delta-agonist-preferring states, whose equilibrium may be regulated by a sulfhydryl group mechanism.
- Published
- 1981
- Full Text
- View/download PDF
14. Correlation of regional brain metabolism with receptor localization during ketamine anesthesia: combined autoradiographic 2-[3H]deoxy-D-glucose receptor binding technique.
- Author
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Hammer, R P, Herkenham, M, Pert, C B, and Quirion, R
- Abstract
LKB film autoradiography of 2-]3H]deoxy-D-glucose uptake shows that ketamine, administered in anesthetic doses, alters the pattern of metabolic activity in rat hippocampus. The labeled metabolic marker can be washed out of the slide-mounted tissue sections by preincubation to permit in vitro autoradiography of drug and neurotransmitter receptors in the same animal. In this way, opiate and phencyclidine receptor distributions may be correlated with patterns of glucose utilization in adjacent sections. If the observed relative enhancement of 2-deoxy-D-glucose uptake in the stratum moleculare of hippocampus reflects elevated metabolism in nerve terminals there, then the binding of ketamine to phencyclidine receptors on neurons in distant afferent sites, such as entorhinal cortex, may initiate the physiologic and metabolic effects.
- Published
- 1982
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15. Phencyclidine (angel dust)/sigma "opiate" receptor: visualization by tritium-sensitive film.
- Author
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Quirion, R, Hammer, R P, Herkenham, M, and Pert, C B
- Abstract
[3H]Phencyclidine ([3H]PCP) binds specifically to an apparently single class of binding sites on slide-mounted sections of rat olfactory bulb (Kd = 46 nM; Bmax = 10.5 fmol per slice). Bound [3H]PCP can be displaced by nonradioactive PCP and a series of its analogs with relative potencies that correlate closely (P less than 0.001) with values determined in a rat discrimination test that utilized PCP as a cue. Although morphine, naloxone, and opiate peptides do not displace bound [3H]PCP, psychotomimetic benzomorphans, classed as "sigma opiates," are quite potent displacers in vitro and have PCP-like behavioral properties in vivo. These results suggest that phencyclidine and the sigma opiates act at the same sites. [3H]PCP binding sites were visualized by using tritium-sensitive LKB film analyzed by computerized densitometry and color coding. The [3H]PCP binds most densely to cortical areas, diffusely in neocortex, and somewhat heterogeneously in the laminae of the hippocampal formation and dentate gyrus. Most of the brainstem and spinal cord show low specific [3H]PCP binding, with gray matter generally showing more binding than white.
- Published
- 1981
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16. The neostriatal mosaic: II. Patch- and matrix-directed mesostriatal dopaminergic and non-dopaminergic systems
- Author
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Gerfen, CR, primary, Herkenham, M, additional, and Thibault, J, additional
- Published
- 1987
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17. Light microscopic localization of brain opiate receptors: a general autoradiographic method which preserves tissue quality
- Author
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Herkenham, M, primary and Pert, CB, additional
- Published
- 1982
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18. Cautionary notes on the use of NF-κB p65 and p50 antibodies for CNS studies
- Author
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Listwak Samuel J, Brown Pierre, Rathore Priyanka, and Herkenham Miles
- Subjects
NF-κB ,transcription factor ,immunohistochemistry ,antibody specificity ,Neurology. Diseases of the nervous system ,RC346-429 - Abstract
Abstract Background The characterization and cellular localization of transcription factors like NF-κB requires the use of antibodies for western blots and immunohistochemistry. However, if target protein levels are low and the antibodies not well characterized, false positive data can result. In studies of NF-κB activity in the CNS, antibodies detecting NF-κB proteins have been used to support the finding that NF-κB is constitutively active in neurons, and activity levels are further increased by neurotoxic treatments, glutamate stimulation, or elevated synaptic activity. The specificity of the antibodies used was analyzed in this study. Methods Selectivity and nonselectivity of commonly used commercial and non-commercial p50 and p65 antibodies were demonstrated in western blot assays conducted in tissues from mutant gene knockout mice lacking the target proteins. Results A few antibodies for p50 and p65 each mark a single band at the appropriate molecular weight in gels containing proteins from wildtype tissue, and this band is absent in proteins from knockout tissues. Several antibodies mark proteins that are present in knockout tissues, indicating that they are nonspecific. These include antibodies raised against the peptide sequence containing the nuclear localization signals of p65 (MAB3026; Chemicon) and p50 (sc-114; Santa Cruz). Some antibodies that recognize target proteins at the correct molecular weight still fail in western blot analysis because they also mark additional proteins and inconsistently so. We show that the criterion for validation by use of blocking peptides can still fail the test of specificity, as demonstrated for several antibodies raised against p65 phosphorylated at serine 276. Finally, even antibodies that show specificity in western blots produce nonspecific neuronal staining by immunohistochemistry. Conclusions We note that many of the findings in the literature about neuronal NF-κB are based on data garnered with antibodies that are not selective for the NF-κB subunit proteins p65 and p50. The data urge caution in interpreting studies of neuronal NF-κB activity in the brain.
- Published
- 2011
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19. Chronic social defeat stress induces meningeal neutrophilia via type I interferon signaling.
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Kigar SL, Lynall ME, DePuyt AE, Atkinson R, Sun VH, Samuels JD, Eassa NE, Poffenberger CN, Lehmann ML, Listwak SJ, Livak F, Elkahloun AG, Clatworthy MR, Bullmore ET, and Herkenham M
- Abstract
Animal models of stress and stress-related disorders are also associated with blood neutrophilia. The mechanistic relevance of this to symptoms or behavior is unclear. We used cytometry, immunohistochemistry, whole tissue clearing, and single-cell sequencing to characterize the meningeal immune response to chronic social defeat (CSD) stress in mice. We find that chronic, but not acute, stress causes meningeal neutrophil accumulation, and CSD increases neutrophil trafficking in vascular channels emanating from skull bone marrow (BM). Transcriptional analysis suggested CSD increases type I interferon (IFN-I) signaling in meningeal neutrophils. Blocking this pathway via the IFN-I receptor (IFNAR) protected against the anhedonic and anxiogenic effects of CSD stress, potentially through reduced infiltration of IFNAR
+ neutrophils into the meninges from skull BM. Our identification of IFN-I signaling as a putative mediator of meningeal neutrophil recruitment may facilitate development of new therapies for stress-related disorders., Competing Interests: Competing interests E.T.B. is a consultant for Sosei Heptares. M.L.L. is currently employed at AstraZeneca but was an employee at NIH at the time this work was conducted. The other authors have no conflicts to declare.- Published
- 2024
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20. Chronic social defeat alters brain vascular-associated cell gene expression patterns leading to vascular dysfunction and immune system activation.
- Author
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Samuels JD, Lotstein ML, Lehmann ML, Elkahloun AG, Banerjee S, and Herkenham M
- Subjects
- Animals, Mice, Immune System, Blood-Brain Barrier, Gene Expression, Social Defeat, Brain
- Abstract
Brain vascular integrity is critical for brain health, and its disruption is implicated in many brain pathologies, including psychiatric disorders. Brain-vascular barriers are a complex cellular landscape composed of endothelial, glial, mural, and immune cells. Yet currently, little is known about these brain vascular-associated cells (BVACs) in health and disease. Previously, we demonstrated that 14 days of chronic social defeat (CSD), a mouse paradigm that produces anxiety and depressive-like behaviors, causes cerebrovascular damage in the form of scattered microbleeds. Here, we developed a technique to isolate barrier-related cells from the mouse brain and subjected the isolated cells to single-cell RNA sequencing. Using this isolation technique, we found an enrichment in BVAC populations, including distinct subsets of endothelial and microglial cells. In CSD compared to non-stress, home-cage control, differential gene expression patterns disclosed biological pathways involving vascular dysfunction, vascular healing, and immune system activation. Overall, our work demonstrates a unique technique to study BVAC populations from fresh brain tissue and suggests that neurovascular dysfunction is a key driver of psychosocial stress-induced brain pathology., (© 2023. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)
- Published
- 2023
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21. The Behavioral Sequelae of Social Defeat Require Microglia and Are Driven by Oxidative Stress in Mice.
- Author
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Lehmann ML, Weigel TK, Poffenberger CN, and Herkenham M
- Subjects
- Acetylcysteine pharmacology, Animals, Antioxidants pharmacology, Male, Mice, Mice, Inbred C57BL, Microglia drug effects, Organic Chemicals toxicity, Receptors, Granulocyte-Macrophage Colony-Stimulating Factor antagonists & inhibitors, Microglia metabolism, Oxidative Stress, Social Behavior, Stress, Psychological metabolism
- Abstract
Chronic social defeat (CSD) in male mice can produce anxiety and aberrant socialization. Animals susceptible to CSD show activation of microglia, which have elevated levels of oxidative stress markers. We hypothesized that microglia and reactive oxygen species (ROS) production contribute to the CSD stress-induced changes in affective behavior. First, we selectively depleted microglia (99%) by administering the CSF1R (colony-stimulating factor 1 receptor) antagonist PLX5622 before and during the 14 d CSD procedure. Microglia-depleted mice in contrast to nondepleted mice were protected from the stress effects measured by light/dark and social interaction tests. ROS production, measured histochemically following dihydroethidium administration, was elevated by CSD, and the production was reduced to basal levels in mice lacking microglia. The deleterious stress effects were also blocked in nondepleted mice by continuous intracerebral administration of N -acetylcysteine (NAC), a ROS inhibitor. In a second experiment, at the end of the CSD period, PLX5622 was discontinued to allow microglial repopulation. After 14 d, the brain had a full complement of newly generated microglia. At this time, the mice that had previously been protected now showed behavioral deficits, and their brain ROS production was elevated, both in all brain cells and in repopulated microglia. NAC administration during repopulation prevented the behavioral decline in the repopulated mice, and it supported behavioral recovery in nondepleted stressed mice. The data suggest that microglia drive elevated ROS production during and after stress exposure. This elevated ROS activity generates a central state supporting dysregulated affect, and it hinders the restoration of behavioral and neurochemical homeostasis after stress cessation. SIGNIFICANCE STATEMENT Chronic psychosocial stress is associated with psychiatric disorders such as depression and anxiety. Understanding the details of CNS cellular contributions to stress effects could lead to the development of intervention strategies. Inflammation and oxidative stress are positively linked to depression severity, but the cellular nature of these processes is not clear. The chronic social defeat (CSD) paradigm in mice produces mood alterations and microglial activation characterized by elevated reactive oxygen species (ROS) production. The depletion of microglia or ROS inhibition prevented adverse stress effects. Microglial repopulation of the brain post-CSD reintroduced adverse stress effects, and ROS inhibition in this phase protected against the effects. The results suggest that stress-induced microglial ROS production drives a central state that supports dysregulated affective behavior., (Copyright © 2019 the authors.)
- Published
- 2019
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22. Decoding microglia responses to psychosocial stress reveals blood-brain barrier breakdown that may drive stress susceptibility.
- Author
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Lehmann ML, Weigel TK, Cooper HA, Elkahloun AG, Kigar SL, and Herkenham M
- Subjects
- Animals, Behavior, Animal, Gene Expression Profiling, Immunohistochemistry, Mice, Microarray Analysis, Blood-Brain Barrier physiopathology, Microglia immunology, Microglia pathology, Stress, Psychological physiopathology
- Abstract
An animal's ability to cope with or succumb to deleterious effects of chronic psychological stress may be rooted in the brain's immune responses manifested in microglial activity. Mice subjected to chronic social defeat (CSD) were categorized as susceptible (CSD-S) or resilient (CSD-R) based on behavioral phenotyping, and their microglia were isolated and analyzed by microarray. Microglia transcriptomes from CSD-S mice were enriched for pathways associated with inflammation, phagocytosis, oxidative stress, and extracellular matrix remodeling. Histochemical experiments confirmed the array predictions: CSD-S microglia showed elevated phagocytosis and oxidative stress, and the brains of CSD-S but not CSD-R or non-stressed control mice showed vascular leakage of intravenously injected fluorescent tracers. The results suggest that the inflammatory profile of CSD-S microglia may be precipitated by extracellular matrix degradation, oxidative stress, microbleeds, and entry and phagocytosis of blood-borne substances into brain parenchyma. We hypothesize that these CNS-centric responses contribute to the stress-susceptible behavioral phenotype.
- Published
- 2018
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23. Chronic social defeat reduces myelination in the mouse medial prefrontal cortex.
- Author
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Lehmann ML, Weigel TK, Elkahloun AG, and Herkenham M
- Subjects
- Animals, Male, Mice, Myelin Sheath metabolism, Myelin Sheath pathology, Prefrontal Cortex metabolism, Prefrontal Cortex pathology, Stress, Psychological metabolism, Stress, Psychological pathology
- Abstract
The medial prefrontal cortex (mPFC) plays a key role in top-down control of the brain's stress axis, and its structure and function are particularly vulnerable to stress effects, which can lead to depression in humans and depressive-like states in animals. We tested whether chronic social defeat produces structural alterations in the mPFC in mice. We first performed a microarray analysis of mPFC gene expression changes induced by defeat, and biological pathway analysis revealed a dominant pattern of down-regulation of myelin-associated genes. Indeed, 69% of the most significantly down-regulated genes were myelin-related. The down regulation was confirmed by in situ hybridization histochemistry for two strongly down-regulated genes, myelin oligodendrocyte glycoprotein (Mog) and ermin (Ermn), and by immunohistochemistry for myelin basic protein. To test for stress-induced changes in myelin integrity, aurophosphate (Black Gold) myelin staining was performed on mPFC sections. Quantitative stereologic analysis showed reduced myelinated fiber length and density. Behavioral analysis confirmed that the 14-day social defeat sessions resulted in induction of depressive-like states measured in social interaction and light/dark tests. The combined data suggest that chronic social defeat induces molecular changes that reduce myelination of the prefrontal cortex, which may be an underlying basis for stress-induced depressive states.
- Published
- 2017
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24. Social defeat induces depressive-like states and microglial activation without involvement of peripheral macrophages.
- Author
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Lehmann ML, Cooper HA, Maric D, and Herkenham M
- Subjects
- Animals, Depression immunology, Depression metabolism, Female, Macrophages immunology, Macrophages metabolism, Male, Mice, Mice, Inbred C57BL, Mice, Transgenic, Microglia immunology, Microglia metabolism, Stress, Psychological metabolism, Stress, Psychological psychology, Depression pathology, Macrophages pathology, Microglia pathology, Social Behavior, Stress, Psychological pathology
- Abstract
Background: We are interested in the causal interactions between psychological stress and activity within different compartments of the immune system. Psychosocial stress has been reported to not only alter microglia morphology but also produce anxiety-like and depressive-like effects by triggering CNS infiltration of macrophages from the periphery. We sought to test these phenomena in a somewhat different but standardized model of chronic social defeat (SD) stress., Methods: We used a paradigm of dyadic home pairing of dominant and subordinate mice that has been validated to induce powerful anxiety-like and depressive-like effects manifested by behavior assessed in social tasks. We administered the SD stress for 3 days (acute SD) or 14 days (chronic SD) and looked for monocyte entry into the brain by three independent means, including CD45 activation states assessed by flow cytometry and tracking fluorescently tagged peripheral cells from Ccr2 (wt/rfp) and Ubc (gfp/gfp) reporter mice. We further characterized the effects of SD stress on microglia using quantitative morphometric analysis, ex vivo phagocytosis assays, flow cytometry, and immunochemistry., Results: We saw no evidence of stress-induced macrophage entry after acute or chronic defeat stress. In comparison, brain infiltration of peripheral cells did occur after endotoxin administration. Furthermore, mutant mice lacking infiltrating macrophages due to CCR2 knockout developed the same degree of chronic SD-induced depressive behavior as wildtype mice. We therefore focused more closely on the intrinsic immune cells, the microglia. Using Cx3cr1 (wt/gpf) microglial reporter mice, we saw by quantitative methods that microglial morphology was not altered by stress at either time point. However, chronic SD mice had elevated numbers of CD68(hi) microglia examined by flow cytometry. CD68 is a marker for phagocytic activity. Indeed, these cells ex vivo showed elevated phagocytosis, confirming the increased activation status of chronic SD microglia. Finally, acute SD but not chronic SD increased microglial proliferation, which occurred selectively in telencephalic stress-related brain areas., Conclusions: In the SD paradigm, changes in CNS-resident microglia numbers and activation states might represent the main immunological component of the psychosocial stress-induced depressive state.
- Published
- 2016
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25. Urine scent marking (USM): a novel test for depressive-like behavior and a predictor of stress resiliency in mice.
- Author
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Lehmann ML, Geddes CE, Lee JL, and Herkenham M
- Subjects
- Animals, Female, Male, Mice, Mice, Inbred C57BL, Territoriality, Behavior, Animal physiology, Depression urine, Stress, Psychological urine
- Abstract
Decreased interest in pleasurable stimuli including social withdrawal and reduced libido are some of the key symptomatic criteria for major depression, and thus assays that measure social and sexual behavior in rodents may be highly appropriate for modeling depressive states. Here we present a novel approach for validating rodent models of depression by assessing male urine scent marking (USM) made in consequence to a spot of urine from a proestrous female. USM is an ethologically important form of sexual communication expressed by males to attract females. The expression of this behavior is highly sensitive and adaptive to environmental cues and social status. We hypothesized that male USM behavior offers a naturalistic measure of social motivation that can be used to evaluate hedonic behaviors relevant to the study of mood disorders. We demonstrated that 1) adult male mice displayed a strong preference for marking proestrous female urine with a high degree of specificity, 2) exposure to chronic social defeat profoundly decreased USM whereas exposure to environmental enrichment increased USM, 3) the standard antidepressant fluoxetine reversed declines in USM induced by social defeat, 4) USM behavior closely correlated with other hedonic measures, and 5) USM scores in non-stressed mice predicted behavioral outcomes after defeat exposure such that mice displaying high preference for marking female urine prior to social defeat showed behavioral resiliency after social defeat. The findings indicate that the USM test is a sensitive, validated measure of psychosocial stress effects that has high predictive value for examination of stress resiliency and vulnerability and their neurobiological substrates.
- Published
- 2013
- Full Text
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26. Cautionary notes on the use of NF-κB p65 and p50 antibodies for CNS studies.
- Author
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Herkenham M, Rathore P, Brown P, and Listwak SJ
- Subjects
- Animals, Hippocampus cytology, Hippocampus metabolism, Mice, Mice, Inbred C57BL, Mice, Knockout, NF-kappa B p50 Subunit genetics, Neurons cytology, Neurons metabolism, Transcription Factor RelA genetics, Antibodies metabolism, Antibody Specificity, Immunohistochemistry methods, NF-kappa B p50 Subunit metabolism, Transcription Factor RelA metabolism
- Abstract
Background: The characterization and cellular localization of transcription factors like NF-κB requires the use of antibodies for western blots and immunohistochemistry. However, if target protein levels are low and the antibodies not well characterized, false positive data can result. In studies of NF-κB activity in the CNS, antibodies detecting NF-κB proteins have been used to support the finding that NF-κB is constitutively active in neurons, and activity levels are further increased by neurotoxic treatments, glutamate stimulation, or elevated synaptic activity. The specificity of the antibodies used was analyzed in this study., Methods: Selectivity and nonselectivity of commonly used commercial and non-commercial p50 and p65 antibodies were demonstrated in western blot assays conducted in tissues from mutant gene knockout mice lacking the target proteins., Results: A few antibodies for p50 and p65 each mark a single band at the appropriate molecular weight in gels containing proteins from wildtype tissue, and this band is absent in proteins from knockout tissues. Several antibodies mark proteins that are present in knockout tissues, indicating that they are nonspecific. These include antibodies raised against the peptide sequence containing the nuclear localization signals of p65 (MAB3026; Chemicon) and p50 (sc-114; Santa Cruz). Some antibodies that recognize target proteins at the correct molecular weight still fail in western blot analysis because they also mark additional proteins and inconsistently so. We show that the criterion for validation by use of blocking peptides can still fail the test of specificity, as demonstrated for several antibodies raised against p65 phosphorylated at serine 276. Finally, even antibodies that show specificity in western blots produce nonspecific neuronal staining by immunohistochemistry., Conclusions: We note that many of the findings in the literature about neuronal NF-κB are based on data garnered with antibodies that are not selective for the NF-κB subunit proteins p65 and p50. The data urge caution in interpreting studies of neuronal NF-κB activity in the brain.
- Published
- 2011
- Full Text
- View/download PDF
27. Three Promoters Regulate Tissue- and Cell Type-specific Expression of Murine Interleukin-1 Receptor Type I.
- Author
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Chen Q, Zhang H, Li Q, An Y, Herkenham M, Lai W, Popovich P, Agarwal S, and Quan N
- Subjects
- Animals, DNA, Complementary genetics, Gene Library, Glucocorticoids genetics, Glucocorticoids metabolism, Interleukin-1 genetics, Mice, Organ Specificity physiology, RNA, Messenger biosynthesis, RNA, Messenger genetics, Receptors, Interleukin-1 genetics, Gene Expression Regulation physiology, Interleukin-1 metabolism, Promoter Regions, Genetic physiology, Receptors, Interleukin-1 biosynthesis
- Abstract
The type 1 interleukin-1 receptor (IL-1R1) mediates diverse functions of interleukin-1 (IL-1) in the nervous, immune, and neuroendocrine systems. It has been suggested previously that the versatile functions of IL-1 may in part be conferred by the multiple promoters of IL-1R1 that have been identified for the human IL-1R1 gene. Promoters for murine IL-1R1 (mIL-1R1) gene have not been studied in detail. We performed 5'-rapid amplification of cDNA ends to determine the transcription start sites (TSS) in mIL-1R1, using mRNAs derived from 24 different tissues. The results revealed three putative TSSs of mIL-1R1. Three full-length cDNAs containing these distinct TSSs were recovered in screens of cloned cDNA libraries. Translation of these cDNAs produced IL-1R1 proteins that were verified by Western blot analysis. IL-1 stimulation of the individual IL-1R1 proteins resulted in the activation of NF-kappaB. Promoter-reporter assay for genomic DNA sequences immediately upstream of the three TSSs validated that the sequences possess promoter activity in a cell type-specific manner. These promoters are termed P1, P2, and P3 of the mIL-1R1, in 5' to 3' order. Quantitative PCR analysis of P1-, P2-, and P3-specific mIL-1R1 mRNAs showed that there is tissue-specific distribution of these mRNAs in vivo, and there are distinct patterns of P1, P2, and P3 mRNA expression in different cell lines. In the brain, P3 mRNA is expressed preferentially in the dentate gyrus. Further, glucocorticoids differentially regulate these promoters in a cell type-specific manner. Together, these results suggest that the different IL-1R1 promoters contribute to the discrete and diverse actions of IL-1.
- Published
- 2009
- Full Text
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28. Bacterial lipopolysaccharide fever is initiated via Toll-like receptor 4 on hematopoietic cells.
- Author
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Steiner AA, Chakravarty S, Rudaya AY, Herkenham M, and Romanovsky AA
- Subjects
- Animals, Bone Marrow Cells cytology, Bone Marrow Cells physiology, Hematopoietic Stem Cells drug effects, Hematopoietic Stem Cells immunology, Mice, Signal Transduction, Fever physiopathology, Hematopoietic Stem Cells physiology, Lipopolysaccharides toxicity, Toll-Like Receptor 4 physiology
- Abstract
Lipopolysaccharide (LPS), a well-known bacterial pyrogen, is recognized by several receptors, including the Toll-like receptor 4 (TLR4), on various cells. Which of these receptors and cells are linked to fever production is unknown. By constructing 4 mouse chimeras and studying their thermoregulatory responses, we found that all 3 phases of the typical LPS fever depend on TLR4 signaling. The first phase is triggered via the TLR4 on hematopoietic cells. The second and third phases involve TLR4 signaling in both hematopoietic and nonhematopoietic cells.
- Published
- 2006
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29. Thermoregulatory responses of rats to conventional preparations of lipopolysaccharide are caused by lipopolysaccharide per se-- not by lipoprotein contaminants.
- Author
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Steiner AA, Chakravarty S, Robbins JR, Dragic AS, Pan J, Herkenham M, and Romanovsky AA
- Subjects
- Animals, Body Temperature drug effects, Drug Contamination, Escherichia coli, Interleukin-1 blood, Interleukin-1 metabolism, Lipoproteins pharmacology, Male, Mice, Mice, Inbred C3H, Rats, Rats, Wistar, Receptors, Immunologic metabolism, Spleen drug effects, Spleen metabolism, Toll-Like Receptor 4, Tumor Necrosis Factor-alpha metabolism, Body Temperature Regulation drug effects, Drug Compounding, Lipopolysaccharides chemistry, Lipopolysaccharides pharmacology
- Abstract
LPS preparations cause a variety of body temperature (T(b)) responses: monophasic fever, different phases of polyphasic fever, and hypothermia. Conventional (c) LPS preparations contain highly active lipoprotein contaminants (endotoxin proteins). Whereas LPS signals predominantly via the Toll-like receptor (TLR) 4, endotoxin proteins signal via TLR2. Several TLR2-dependent responses of immunocytes to cLPS in vitro are triggered by endotoxin proteins and not by LPS itself. We tested whether any T(b) response to cLPS from Escherichia coli 055:B5 is triggered by non-TLR4-signaling contaminants. A decontaminated (d) LPS preparation (free of endotoxin proteins) was produced by subjecting cLPS to phenol-water reextraction. The presence of non-TLR4-signaling contaminants in cLPS (and their absence in dLPS) was confirmed by showing that cLPS (but not dLPS) induced IL-1beta expression in the spleen and increased serum levels of TNF-alpha and IL-1beta of C3H/HeJ mice; these mice bear a nonfunctional TLR4. Yet, both cLPS and dLPS caused cytokine responses in C3H/HeOuJ mice; these mice bear a fully functional TLR4. We then studied the T(b) responses to cLPS and dLPS in Wistar rats preimplanted with jugular catheters. At a neutral ambient temperature (30 degrees C), a low (0.1 microg/kg iv) dose of cLPS caused a monophasic fever, whereas a moderate (10 microg/kg iv) dose produced a polyphasic fever. In the cold (20 degrees C), a high (500 microg/kg iv) dose of cLPS caused hypothermia. All T(b) responses to dLPS were identical to those of cLPS. We conclude that all known T(b) responses to LPS preparations are triggered by LPS per se and not by non-TLR4-signaling contaminants of such preparations.
- Published
- 2005
- Full Text
- View/download PDF
30. Toll-like receptor 4 on nonhematopoietic cells sustains CNS inflammation during endotoxemia, independent of systemic cytokines.
- Author
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Chakravarty S and Herkenham M
- Subjects
- Animals, Bone Marrow Transplantation, CX3C Chemokine Receptor 1, Cell Lineage, Choroid Plexus metabolism, Corticosterone blood, Cytokines physiology, Encephalitis metabolism, Encephalitis pathology, Endotoxemia metabolism, Endotoxemia pathology, Gene Expression Regulation, Hypothalamo-Hypophyseal System physiopathology, I-kappa B Proteins biosynthesis, I-kappa B Proteins genetics, Lipopolysaccharides toxicity, Male, Meninges metabolism, Mice, Mice, Inbred C3H, Mice, Inbred C57BL, NF-KappaB Inhibitor alpha, Pituitary-Adrenal System physiopathology, RNA, Messenger biosynthesis, RNA, Messenger genetics, Radiation Chimera, Receptors, Chemokine biosynthesis, Receptors, Chemokine genetics, Receptors, Immunologic biosynthesis, Receptors, Immunologic deficiency, Receptors, Immunologic genetics, Spleen metabolism, Toll-Like Receptor 4, Brain metabolism, Encephalitis etiology, Endotoxemia complications, Receptors, Immunologic physiology
- Abstract
Inflammatory agonists such as lipopolysaccharide (LPS) induce robust systemic as well as CNS responses after peripheral administration. Responses in the innate immune system require triggering of toll-like receptor 4 (TLR4), but the origin of CNS sequelas has been controversial. We demonstrate expression of TLR4 transcripts in mouse brain in the meninges, ventricular ependyma, circumventricular organs, along the vasculature, and in parenchymal microglia. The contribution of TLR4 expressed in CNS resident versus hematopoietic cells to the development of CNS inflammation was examined using chimeric mice. Reciprocal bone marrow chimeras between wild-type and TLR4 mutant mice show that TLR4 on CNS resident cells is critically required for sustained inflammation in the brain after systemic LPS administration. Hematopoietic TLR4 alone supported the systemic release of acute phase cytokines, but transcription of proinflammatory genes in the CNS was reduced in duration. In contrast, TLR4 function in radiation-resistant cells was sufficient for inflammatory progression in the brains of chimeric mice, despite the striking absence of cytokine elevations in serum. Surprisingly, a temporal rise in serum corticosterone was also dependent on TLR4 signaling in nonhematopoietic cells. Our findings demonstrate a requirement for TLR4 function in CNS-resident cells, independent of systemic cytokine effects, for sustained CNS-specific inflammation and corticosterone rise during endotoxemia.
- Published
- 2005
- Full Text
- View/download PDF
31. Folliculo-stellate (FS) cells of the anterior pituitary mediate interactions between the endocrine and immune systems.
- Author
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Herkenham M
- Subjects
- Animals, Macrophage Migration-Inhibitory Factors physiology, Pituitary Gland, Anterior cytology, Endocrine Glands physiology, Immune System physiology, Pituitary Gland, Anterior physiology
- Published
- 2005
- Full Text
- View/download PDF
32. Hyperforin-containing extracts of St John's wort fail to alter gene transcription in brain areas involved in HPA axis control in a long-term treatment regimen in rats.
- Author
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Butterweck V, Winterhoff H, and Herkenham M
- Subjects
- Adrenal Glands drug effects, Adrenocorticotropic Hormone blood, Animals, Body Weight drug effects, Brain anatomy & histology, Brain metabolism, Bridged Bicyclo Compounds, Corticosterone blood, Drug Interactions, Hypothalamo-Hypophyseal System metabolism, Immunohistochemistry, In Situ Hybridization methods, Male, Organ Size drug effects, Phloroglucinol analogs & derivatives, Phosphoproteins genetics, Phosphoproteins metabolism, Pituitary-Adrenal System metabolism, Plant Extracts pharmacology, Pro-Opiomelanocortin genetics, Pro-Opiomelanocortin metabolism, RNA, Messenger metabolism, Radioimmunoassay methods, Rats, Receptor, Serotonin, 5-HT1A metabolism, Receptors, Glucocorticoid metabolism, Receptors, Mineralocorticoid metabolism, Time Factors, Tyrosine 3-Monooxygenase metabolism, Anti-Bacterial Agents pharmacology, Antidepressive Agents pharmacology, Brain drug effects, Hypericum chemistry, Hypothalamo-Hypophyseal System drug effects, Pituitary-Adrenal System drug effects, Terpenes pharmacology, Transcription, Genetic drug effects
- Abstract
We previously showed that a methanolic extract of St John's wort (SJW) (Hypericum) and hypericin, one of its active constituents, both have delayed regulation of genes that are involved in the control of the hypothalamic-pituitary-adrenal (HPA) axis. Hyperforin, another constituent of SJW, is active in vitro and has been proposed to be the active constituent for therapeutic efficacy in depression. We therefore examined if hyperforin has delayed effects on HPA axis control centers similar to those of Hypericum and hypericin. We used in situ hybridization histochemistry to examine in rats the effects of short-term (2 weeks) and long-term (8 weeks) oral administration of two hyperforin preparations, fluoxetine (positive control), and haloperidol (negative control) on the expression of genes involved in the regulation of the HPA axis. Fluoxetine (10 mg/kg) given daily for 8 weeks, but not 2 weeks, significantly decreased levels of corticotropin-releasing hormone (CRH) mRNA by 22% in the paraventricular nucleus (PVN) of the hypothalamus and tyrosine hydroxylase (TH) mRNA by 23% in the locus coeruleus. Fluoxetine increased levels of mineralocorticoid (MR) (17%), glucocorticoid (GR) (18%), and 5-HT(1A) receptor (21%) mRNAs in the hippocampus at 8, but not 2, weeks. Comparable to haloperidol (1 mg/kg), neither the hyperforin-rich CO(2) extract (27 mg/kg) nor hyperforin-trimethoxybenzoate (8 mg/kg) altered mRNA levels in brain structures relevant for HPA axis control at either time point. These data suggest that hyperforin and hyperforin derivatives are not involved in the regulation of genes that control HPA axis function.
- Published
- 2003
- Full Text
- View/download PDF
33. Induction of IkappaBalpha mRNA expression in the brain by glucocorticoids: a negative feedback mechanism for immune-to-brain signaling.
- Author
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Quan N, He L, Lai W, Shen T, and Herkenham M
- Subjects
- Animals, Brain drug effects, Brain immunology, Feedback, Gene Expression Regulation physiology, Glucocorticoids pharmacology, Immune System physiology, Lipopolysaccharides pharmacology, Male, NF-KappaB Inhibitor alpha, NF-kappa B antagonists & inhibitors, RNA, Messenger genetics, Rats, Rats, Sprague-Dawley, Signal Transduction, Transcription, Genetic physiology, Brain physiology, DNA-Binding Proteins genetics, Dexamethasone pharmacology, Gene Expression Regulation drug effects, I-kappa B Proteins, Interleukin-1 genetics, Transcription, Genetic drug effects
- Abstract
Peripheral injection of bacterial endotoxin lipopolysaccharide (LPS) induces brain mRNA expression of the proinflammatory cytokines interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha and the cytokine-responsive immediate-early gene IkappaBalpha. Peripheral LPS also increases levels of plasma glucocorticoids. Whether the induction of IkappaBalpha mRNA in the brain after peripheral LPS injection is caused by the feedback action of glucocorticoids has not been determined. In this study, we examined the mRNA expression of IkappaBalpha and IL-1beta in the rat brain by in situ hybridization histochemistry. Injection of the glucocorticoid agonist dexamethasone induced IkappaBalpha mRNA expression in the brain in a pattern identical to that of LPS injection. LPS but not dexamethasone also induced IL-1beta mRNA expression. Pretreatment with dexamethasone 30 min before LPS injection enhanced the expression of IkappaBalpha mRNA in the brain in a dose-dependent manner. Immobilization of rats for 2 hr (which raises glucocorticoid levels) also induced IkappaBalpha mRNA expression without inducing the expression of IL-1beta. Brain IkappaBalpha expression induced by peripheral LPS injection was attenuated by pretreatment of rats with the glucocorticoid antagonist RU-486. Finally, increased expression of IL-1beta mRNA in the brain was observed at 4 hr after peripheral LPS injection in adrenalectomized rats compared with sham-operated rats. These results reveal that in the brain glucocorticoids selectively induce IkappaBalpha mRNA expression, which serves as a negative feedback mechanism for peripheral LPS-induced synthesis of proinflammatory cytokines. Such an inhibitory control mechanism may be important for preventing prolonged expression of proinflammatory cytokines in the brain after peripheral immune challenge.
- Published
- 2000
34. Increased mortality, hypoactivity, and hypoalgesia in cannabinoid CB1 receptor knockout mice.
- Author
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Zimmer A, Zimmer AM, Hohmann AG, Herkenham M, and Bonner TI
- Subjects
- Animals, Body Temperature drug effects, Brain metabolism, Central Nervous System drug effects, Cyclohexanols metabolism, Dronabinol analogs & derivatives, Gene Targeting methods, Mice, Mice, Knockout, Molecular Structure, Motor Activity drug effects, Receptors, Cannabinoid, Behavior, Animal drug effects, Dronabinol pharmacology, Receptors, Drug genetics
- Abstract
Delta9-Tetrahydrocannabinol (Delta9-THC), the major psychoactive ingredient in preparations of Cannabis sativa (marijuana, hashish), elicits central nervous system (CNS) responses, including cognitive alterations and euphoria. These responses account for the abuse potential of cannabis, while other effects such as analgesia suggest potential medicinal applications. To study the role of the major known target of cannabinoids in the CNS, the CB1 cannabinoid receptor, we have produced a mouse strain with a disrupted CB1 gene. CB1 knockout mice appeared healthy and fertile, but they had a significantly increased mortality rate. They also displayed reduced locomotor activity, increased ring catalepsy, and hypoalgesia in hotplate and formalin tests. Delta9-THC-induced ring-catalepsy, hypomobility, and hypothermia were completely absent in CB1 mutant mice. In contrast, we still found Delta9-THC-induced analgesia in the tail-flick test and other behavioral (licking of the abdomen) and physiological (diarrhea) responses after Delta9-THC administration. Thus, most, but not all, CNS effects of Delta9-THC are mediated by the CB1 receptor.
- Published
- 1999
- Full Text
- View/download PDF
35. Induction of inhibitory factor kappaBalpha mRNA in the central nervous system after peripheral lipopolysaccharide administration: an in situ hybridization histochemistry study in the rat.
- Author
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Quan N, Whiteside M, Kim L, and Herkenham M
- Subjects
- Animals, Central Nervous System metabolism, DNA-Binding Proteins genetics, In Situ Hybridization, Lipopolysaccharides administration & dosage, Male, NF-KappaB Inhibitor alpha, RNA, Messenger genetics, RNA, Messenger metabolism, Rats, Rats, Sprague-Dawley, Central Nervous System drug effects, DNA-Binding Proteins biosynthesis, I-kappa B Proteins, Lipopolysaccharides pharmacology
- Abstract
In this study we investigate the mRNA expression of inhibitory factor kappaBalpha (IkappaBalpha) in cells of the rat brain induced by an intraperitoneal (i.p.) injection of lipopolysaccharide (LPS). IkappaB controls the activity of nuclear factor kappaB, which regulates the transcription of many immune signal molecules. The detection of IkappaB induction, therefore, would reveal the extent and the cellular location of brain-derived immune molecules in response to peripheral immune challenges. Low levels of IkappaBalpha mRNA were found in the large blood vessels and in circumventricular organs (CVOs) of saline-injected control animals. After an i.p. LPS injection (2.5 mg/kg), dramatic induction of IkappaBalpha mRNA occurred in four spatio-temporal patterns. Induced signals were first detected at 0.5 hr in the lumen of large blood vessels and in blood vessels of the choroid plexus and CVOs. Second, at 1-2 hr, labeling dramatically increased in the CVOs and choroid plexus and spread to small vascular and glial cells throughout the entire brain; these responses peaked at 2 hr and declined thereafter. Third, cells of the meninges became activated at 2 hr and persisted until 12 hr after the LPS injection. Finally, only at 12 hr, induced signals were present in ventricular ependyma. Thus, IkappaBalpha mRNA is induced in brain after peripheral LPS injection, beginning in cells lining the blood side of the blood-brain barrier and progressing to cells inside brain. The spatiotemporal patterns suggest that cells of the blood-brain barrier synthesize immune signal molecules to activate cells inside the central nervous system in response to peripheral LPS. The cerebrospinal fluid appears to be a conduit for these signal molecules.
- Published
- 1997
- Full Text
- View/download PDF
36. Opiate receptor distribution in the cerebral cortex of the Rhesus monkey.
- Author
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Wise SP and Herkenham M
- Subjects
- Animals, Autoradiography, Brain Mapping, Cerebral Cortex cytology, Macaca mulatta, Naloxone metabolism, Cerebral Cortex metabolism, Receptors, Opioid metabolism
- Abstract
The distribution of opiate receptors in the cerebral cortex of the rhesus monkey (Macaca mulatta) was determined by autoradiographic visualization of [3H]naloxone binding to tissue sections. Naloxone was bound in relatively large amounts to the cortical laminae containing the cell bodies of output neurons, to a varying set of additional laminae in different cortical fields, to fields closer to more primitive types of cortex, and to polysensory cortical fields. From these laminar and areal variations in distribution, it appears that opiate receptors play a role in specific aspects of cortical function.
- Published
- 1982
- Full Text
- View/download PDF
37. Laminar organization of thalamic projections to the rat neocortex.
- Author
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Herkenham M
- Subjects
- Animals, Autoradiography, Brain Mapping, Neural Pathways cytology, Phylogeny, Rats, Cerebral Cortex cytology, Thalamic Nuclei cytology
- Abstract
Nerve fibers transmitting information from the thalamus to the cerebral cortex may be classified according to their major cortical layers of termination. (i) One class consists of inputs from thalamic relay nuclei for vision, audition, and somesthesis to layer IV, layer III, or both. In contrast, autoradiographic studies of projections from other thalamic nuclei reveal strikingly different patterns of termination: (ii) layer VI (or layer V, or both) is the target of fibers from the intralaminar nuclei, and (iii) layer I is the target for fibers from the ventromedial and magnocellular medial geniculate nuclei. (iv) The remaining class is typified by termination both in layer I and in additional layers that depend on the cortical area in which the terminations are found. The data demonstrate that convergent thalamic inputs to a given cortical area are usually not confluent within a layer and provide a new frame-work for categorizing thalamic nuclei.
- Published
- 1980
- Full Text
- View/download PDF
38. Mosaic distribution of opiate receptors, parafascicular projections and acetylcholinesterase in rat striatum.
- Author
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Herkenham M and Pert CB
- Subjects
- Animals, Enkephalin, Leucine-2-Alanine, Enkephalins metabolism, Etorphine metabolism, Leucine metabolism, Naloxone metabolism, Proline metabolism, Rats, Acetylcholinesterase metabolism, Axonal Transport, Caudate Nucleus metabolism, Corpus Striatum metabolism, Receptors, Opioid metabolism
- Published
- 1981
- Full Text
- View/download PDF
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