Clozapine N-oxide
Chemical Name: 8-Chloro-11-(4-methyl-4-oxido-1-piperazinyl)-5H-dibenzo[b,e][1,4]diazepine
Purity: ≥99%
Biological Activity
Clozapine N-oxide is a synthetic ligand for human muscarinic engineered receptors, designer receptors activated by designer drugs (DREADD). Binds and activates hM3Dq and hM4Di DREADDs in vitro and in vivo. Metabolite of Clozapine (Cat. No. 0444). Shown to be a P-glycoprotein (P-gp) efflux pump substrate.Example Applications of Clozapine N-oxide (CNO):
- Silences hippocampal neurons expressing hM4Di DREADDs in vitro
- Inhibits locomotor activity of mice expressing hM3Dq in GABAergic VTA neurons
- Inhibits short term memory retrieval in mice expressing hM4Di in hippocampal neurons
- Induces heat and mechanical hypersensitivity in mice expressing hM4Di in enkephalinergic neurons in the right lumbar dorsal horn
- Inhibits markers of social aggression in mice expressing hM4Di in hippocampal CA2 neurons
Alternative potent and selective hM3Dq and hM4Di DREADD agonists available: Deschloroclozapine and DREADD agonist 21
Water soluble salt also available.
This product contains 1 molar equivalent of ethanol.
Technical Data
The technical data provided above is for guidance only.
For batch specific data refer to the Certificate of Analysis.
Tocris products are intended for laboratory research use only, unless stated otherwise.
Background References
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Gs-coupled GPCR signalling in AgRP neurons triggers sustained increase in food intake.
Nakajima et al.
Nat. Commun., 2016;8:10268 -
Evolving the lock to fit the key to create a family of G protein-coupled receptors potently activated by an inert ligand.
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LMO4 is essential for paraventricular hypothalamic neuronal activity and calcium channel expression to prevent hyperphagia.
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DREADDs for Neuroscientists.
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Neuron, 2016;89:683 -
The involvement of CYP1A2 and CYP3A4 in the metabolism of cloz.
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Br.J.Clin.Pharmacol., 1997;44:439 -
Chemogenetics revealed: DREADD occupancy and activation via converted cloz.
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Science, 2017;357:503 -
Adenosine triphosphate is co-secreted with glucagon-like peptide-1 to modulate intestinal enterocytes and afferent neurons
VB Lu, J Rievaj, EA O'Flaherty, CA Smith, R Pais, LA Pattison, G Tolhurst, AB Leiter, DC Bulmer, FM Gribble, F Reimann
Nat Commun, 2019;10(1):1029.
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Citations for Clozapine N-oxide
The citations listed below are publications that use Tocris products. Selected citations for Clozapine N-oxide include:
139 Citations: Showing 1 - 10
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Arc-Expressing Accessory Olfactory Bulb Interneurons Support Chemosensory Social Behavioral Plasticity.
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J Neurosci 2023;43:1178-1190
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Transient Receptor Potential Ankyrin-1-expressing vagus nerve fibers mediate IL-1� induced hypothermia and reflex anti-inflammatory responses
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Mol Med 2023;29:4
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Presynaptic Gq-coupled receptors drive biphasic dopamine transporter trafficking that modulates dopamine clearance and motor function.
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J Biol Chem 2023;299:102900
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Norepinephrine links astrocytic activity to regulation of cortical state.
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Nat Neurosci 2023;26:579-593
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Serotonergic neurons control cortical neuronal intracellular energy dynamics by modulating astrocyte-neuron lactate shuttle.
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iScience 2023;26:105830
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DDC-Promoter-Driven Chemogenetic Activation of SNpc Dopaminergic Neurons Alleviates Parkinsonian Motor Symptoms.
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Int J Mol Sci 2023;24
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Denervated mouse CA1 pyramidal neurons express homeostatic synaptic plasticity following entorhinal cortex lesion.
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Front Mol Neurosci 2023;16:1148219
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Orexinergic neurons modulate stress coping responses in mice.
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Auts2 deletion involves in DG hypoplasia and social recognition deficit: The developmental and neural circuit mechanisms.
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Sci Adv 2022;8:eabk1238
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Mapping and targeted viral activation of pancreatic nerves in mice reveal their roles in the regulation of glucose metabolism.
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Nat Biomed Eng 2022;6:1298-1316
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High fat diet blunts stress-induced hypophagia and activation of Glp1r dorsal lateral septum neurons in male but not in female mice.
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Mol Metab 2022;64:101571
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Distinct Contribution of Granular and Agranular Subdivisions of the Retrosplenial Cortex to Remote Contextual Fear Memory Retrieval.
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J Neurosci 2022;42:877-893
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Dynamic interplay between thalamic activity and Cajal-Retzius cells regulates the wiring of cortical layer 1.
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Cell Rep 2022;39:110667
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Hypothalamic pregnenolone mediates recognition memory in the context of metabolic disorders.
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Cell Metab 2022;34:269-284.e9
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Hypothalamic AgRP neurons exert top-down control on systemic TNF-α release during endotoxemia.
Authors: Simon M Et al.
Curr Biol 2022;32:4699-4706.e4
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Median raphe region GABAergic neurons contribute to social interest in mouse.
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Life Sci 2022;289:120223
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High-fat diet induces depression-like phenotype via astrocyte-mediated hyperactivation of ventral hippocampal glutamatergic afferents to the nucleus accumbens.
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Mol Psychiatry 2022;27:4372-4384
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Why do mice squeak? Toward a better understanding of defensive vocalization.
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TRAPing Ghrelin-Activated Circuits: A Novel Tool to Identify, Target and Control Hormone-Responsive Populations in TRAP2 Mice.
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Int J Mol Sci 2022;23
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Microglia modulate blood flow, neurovascular coupling, and hypoperfusion via purinergic actions.
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J Exp Med 2022;219
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Control of mammalian locomotion by ventral spinocerebellar tract neurons.
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Cell 2022;185:328-344.e26
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Parvalbumin Neurons in Zona Incerta Regulate Itch in Mice.
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Front Mol Neurosci 2022;15:843754
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Targeted activation of midbrain neurons restores locomotor function in mouse models of parkinsonism.
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Nat Commun 2022;13:504
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Chemogenetic stimulation of the Gi pathway in astrocytes suppresses neuroinflammation.
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Pharmacol Res Perspect 2021;9:e00822
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A neural substrate of compulsive alcohol use.
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Sci Adv 2021;7
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Presymptomatic training mitigates functional deficits in a mouse model of Rett syndrome.
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Nature 2021;592:596-600
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A functional map for diverse forelimb actions within brainstem circuitry.
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Nature 2021;590:445-450
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Somatostatin neurons of the bed nucleus of stria terminalis enhance associative fear memory consolidation in mice.
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J Neurosci 2021;41:1982
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Projections from Infralimbic Cortex to Paraventricular Thalamus Mediate Fear Extinction Retrieval.
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Neurosci Bull 2021;37:229-241
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Lateral hypothalamic LEPR neurons drive appetitive but not consummatory behaviors.
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Cell Rep 2021;36:109615
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Identification of a population of peripheral sensory neurons that regulates blood pressure.
Authors: Morelli Et al.
Cell Rep 2021;35
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Top-down control of conditioned overconsumption is mediated by insular cortex Nos1 neurons.
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Cell Metab 2021;33:1418-1432.e6
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Thalamic input to motor cortex facilitates goal-directed action initiation.
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Curr Biol 2021;31:4148-4155.e4
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Hypothalamic control of interoceptive hunger.
Authors: Sarah Et al.
Curr Biol 2021;31:3797-3809.e5
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Oxytocin shapes spontaneous activity patterns in the developing visual cortex by activating somatostatin interneurons.
Authors: Maldonado Et al.
Curr Biol 2021;31:322
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Ventro-dorsal Hippocampal Pathway Gates Novelty-Induced Contextual Memory Formation.
Authors: Ryuichi Et al.
Curr Biol 2021;31:25-38.e5
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Enhancing mask activity in dopaminergic neurons extends lifespan in flies.
Authors: Xiaolin Et al.
Aging Cell 2021;20:e13493
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Neurochemically distinct populations of the bed nucleus of stria terminalis modulate innate fear response to weak threat evoked by predator odor stimuli.
Authors: Máté Et al.
Neurobiol Stress 2021;15:100415
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Chronic chemogenetic stimulation of the anterior olfactory nucleus reduces newborn neuron survival in the adult mouse olfactory bulb.
Authors: Chris Et al.
J Neurochem 2021;158:1186-1198
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Gut-innervating TRPV1+ Neurons Drive Chronic Visceral Pain via Microglial P2Y12 Receptor.
Authors: Gerald W Et al.
Cell Mol Gastroenterol Hepatol 2021;13:977-999
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Hunger-promoting AgRP neurons trigger an astrocyte-mediated feed-forward autoactivation loop in mice.
Authors: Luis Et al.
J Clin Invest 2021;131
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PDX1LOW MAFALOW β-cells contribute to islet function and insulin release.
Authors: Qiao Et al.
Nat Commun 2021;12:674
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NTS Prlh overcomes orexigenic stimuli and ameliorates dietary and genetic forms of obesity.
Authors: David P Et al.
Nat Commun 2021;12:5175
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Locomotor deficits in a mouse model of ALS are paralleled by loss of V1-interneuron connections onto fast motor neurons.
Authors: Peter Et al.
Nat Commun 2021;12:3251
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Efferent feedback controls bilateral auditory spontaneous activity.
Authors: Lei Et al.
Nat Commun 2021;12:2449
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Specific and behaviorally consequential astrocyte Gq GPCR signaling attenuation in vivo with iβARK.
Authors: Giovanni Et al.
Neuron 2021;109:2256-2274.e9
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Loss of nigral excitation of cholinergic interneurons contributes to parkinsonian motor impairments.
Authors: Jason Et al.
Neuron 2021;109:1137-1149.e5
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An excitatory lateral hypothalamic circuit orchestrating pain behaviors in mice.
Authors: Alfred J Et al.
Elife 2021;10
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A bidirectional corticoamygdala circuit for the encoding and retrieval of detailed reward memories.
Authors: Carlos Et al.
Elife 2021;10
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Loss of cortical control over the descending pain modulatory system determines the development of the neuropathic pain state in rats.
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Elife 2021;10
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ARCGHR Neurons Regulate Muscle Glucose Uptake.
Authors: Alan C Et al.
Cells 2021;10
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tTARGIT AAVs mediate the sensitive and flexible manipulation of intersectional neuronal populations in mice.
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Elife 2021;10
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Corticothalamic gating of population auditory thalamocortical transmission in mouse.
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Elife 2021;10
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Chronic postnatal chemogenetic activation of forebrain excitatory neurons evokes persistent changes in mood behavior
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ELife 2020;9
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Defective memory engram reactivation underlies impaired fear memory recall in Fragile X syndrome.
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Elife 2020;9
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Probabilistic, spinally-gated control of bladder pressure and autonomous micturition by Barrington's nucleus CRH neurons.
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Elife 2020;9
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A limbic circuit selectively links active escape to food suppression.
Authors: Lisa E Et al.
Elife 2020;9
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Chronic hM3Dq signaling in microglia ameliorates neuroinflammation in male mice.
Authors: Wei-Yang Et al.
Brain Behav Immun 2020;88:791-801
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Modular, Circuit-Based Interventions Rescue Hippocampal-Dependent Social and Spatial Memory in a 22q11.2 Deletion Syndrome Mouse Model.
Authors: Stewart A Et al.
Biol Psychiatry 2020;88:710-718
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Value-guided remapping of sensory cortex by lateral orbitofrontal cortex.
Authors: Jasper Et al.
Nature 2020;585:245-250
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In vivo electrophysiological validation of DREADD-based inhibition of pallidal neurons in the non-human primate.
Authors: Deffains Et al.
Eur J Neurosci 2020;53:2192
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Leptin receptor-expressing nucleus tractus solitarius neurons suppress food intake independently of GLP1 in mice.
Authors: Martin G Et al.
JCI Insight 2020;5
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Dynamic Changes in the Bridging Collaterals of the Basal Ganglia Circuitry Control Stress-Related Behaviors in Mice.
Authors: Se-Young Et al.
Mol Cells 2020;43:360-372
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Developmental Phase Transitions in Spatial Organization of Spontaneous Activity in Postnatal Barrel Cortex Layer 4.
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J Neurosci 2020;40:7637-7650
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Chemogenetic Modulation and Single-Photon Calcium Imaging in Anterior Cingulate Cortex Reveal a Mechanism for Effort-Based Decisions
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J Neurosci 2020;40:5628
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Specific Hippocampal Interneurons Shape Consolidation of Recognition Memory.
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Cell Rep 2020;32:108046
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Social Behavior Is Modulated by Valence-Encoding mPFC-Amygdala Sub-circuitry.
Authors: Wen-Chin Et al.
Cell Rep 2020;32:107899
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Calcitonin Receptor Neurons in the Mouse Nucleus Tractus Solitarius Control Energy Balance via the Non-aversive Suppression of Feeding.
Authors: Martin G Et al.
Cell Metab 2020;31:301-312.e5
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Brain Circuit of Claustrophobia-like Behavior in Mice Identified by Upstream Tracing of Sighing.
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Cell Rep 2020;31:107779
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A Sparse, Spatially Biased Subtype of Mature Granule Cell Dominates Recruitment in Hippocampal-Associated Behaviors.
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Cell Rep 2020;31:107551
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Gαs directly drives PDZ-RhoGEF signaling to Cdc42.
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J Biol Chem 2020;295:16920-16928
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Brainstem neurons that command mammalian locomotor asymmetries.
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Nat Neurosci 2020;23:730
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Enhancement of Drug Seeking Following Drug Taking in a Sexual Context Requires Anterior Cingulate Cortex Activity in Male Rats.
Authors: Lique M Et al.
Front Behav Neurosci 2020;14:87
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Neuronal activity modulates alpha-synuclein aggregation and spreading in organotypic brain slice cultures and in vivo.
Authors: John Q Et al.
Acta Neuropathol 2020;140:831-849
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Activation of G-Protein Coupled Receptor-Gαi Signaling Increases Keratinocyte Proliferation and Reduces Differentiation, Leading to Epidermal Hyperplasia.
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J Invest Dermatol 2020;140:1195-1203.e3
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Gabapentinoid treatment promotes corticospinal plasticity and regeneration following murine spinal cord injury.
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J Clin Invest 2020;130:345-358
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Anatomically and functionally distinct locus coeruleus efferents mediate opposing effects on anxiety-like behavior
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Neurobiology of Stress 2020;13
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Distinct CCK-positive SFO neurons are involved in persistent or transient suppression of water intake.
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Nat Commun 2020;11:5692
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Polysynaptic inhibition between striatal cholinergic interneurons shapes their network activity patterns in a dopamine-dependent manner.
Authors: Dorst Et al.
Nat Commun 2020;11:5113
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Dissociable mesolimbic dopamine circuits control responding triggered by alcohol-predictive discrete cues and contexts.
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Nat Commun 2020;11:3764
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Context-Specific Striatal Astrocyte Molecular Responses Are Phenotypically Exploitable.
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Neuron 2020;108:1146-1162.e10
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Region-Specific Transcriptional Control of Astrocyte Function Oversees Local Circuit Activities.
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Neuron 2020;106:992-1008.e9
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Astroglia in lateral habenula is essential for antidepressant efficacy of light
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BioRXiv - not yet peer reviewed 2020;
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Sodium regulates clock time and output via an excitatory GABAergic pathway.
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Nature 2020;
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Distinct roles of striatal direct and indirect pathways in value-based decision making.
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Elife 2019;8
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Anterior cingulate cortex modulates the affective-motivative dimension of hyperosmolality-induced thirst.
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J Physiol 2019;597:4851
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Prefrontal Theta Oscillations Promote Selective Encoding of Behaviorally Relevant Events.
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Eneuro 2019;5
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Galanin Neurons Unite Sleep Homeostasis and α2-Adrenergic Sedation
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Curr Biol 2019;29:3315
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Oxytocin Signaling in the Central Amygdala Modulates Emotion Discrimination in Mice.
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Curr Biol 2019;29:1938-1953.e6
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Gβγ signaling to the chemotactic effector P-REX1 and mammalian cell migration is directly regulated by Gαq and Gα13 proteins.
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J Biol Chem 2019;294:531-546
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Maladaptive Downregulation of Autonomous Subthalamic Nucleus Activity following the Loss of Midbrain Dopamine Neurons
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Cell Rep 2019;28:992
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Recruitment of Spinoparabrachial Neurons by Dorsal Horn Calretinin Neurons.
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Cell Rep 2019;28:1429-1438.e4
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Activation of Astrocytic μ-Opioid Receptor Causes Conditioned Place Preference
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Cell Rep 2019;28:1154
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Routing Hippocampal Information Flow through Parvalbumin Interneuron Plasticity in Area CA2.
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Cell Rep 2019;27:86
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Illuminating G-Protein-Coupling Selectivity of GPCRs.
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Cell 2019;177:1933-1947.e25
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Hyperactivity with Disrupted Attention by Activation of an Astrocyte Synaptogenic Cue.
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Cell 2019;177:1280-1292.e20
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Remotely controlled chemomagnetic modulation of targeted neural circuits.
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Nat Nanotechnol 2019;14:967-973
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Chemogenetic Activation of Adrenocortical Gq Signaling Causes Hyperaldosteronism and Disrupts Functional Zonation.
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J Clin Invest 2019;130:83
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Cholinergic neural activity directs retinal layer-specific angiogenesis and blood retinal barrier formation.
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Nat Commun 2019;10:2477
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Adenosine triphosphate is co-secreted with glucagon-like peptide-1 to modulate intestinal enterocytes and afferent neurons.
Authors: Lu Et al.
Nat.commun. 2019;10:1029
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Natural and Drug Rewards Engage Distinct Pathways that Converge on Coordinated Hypothalamic and Reward Circuits.
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Neuron 2019;103:891-908.e6
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A hippocampal circuit linking dorsal CA2 to ventral CA1 critical for social memory dynamics.
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Nat.Commun. 2018;9:4163
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Reducing Astrocyte Calcium Signaling In Vivo Alters Striatal Microcircuits and Causes Repetitive Behavior.
Authors: Christopher J Et al.
Neuron 2018;99:1170-1187.e9
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The role of calretinin-expressing granule cells in olfactory bulb functions and odor behavior.
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Sci Rep 2018;8:9385
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Dynamic interaction of local and transhemispheric networks is necessary for progressive intensification of hippocampal seizures.
Authors: Berglind Et al.
Sci Rep 2018;8:5669
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Cholinergic neurons in medial septum maintain anxiety-like behaviors induced by chronic inflammatory pain.
Authors: Jiang Et al.
Neurosci Letts 2018;671:7
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A circuit from hippocampal CA2 to lateral septum disinhibits social aggression.
Authors: Leroy Et al.
Nature 2018;564:213
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Parallel Arousal Pathways in the Lateral Hypothalamus.
Authors: Heiss Et al.
Eneuro 2018;5
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Kisspeptin neurones in the posterodorsal medial amygdala modulate sexual partner preference and anxiety in male mice.
Authors: Adekunbi Et al.
J Neuroendocrinol 2018;30:e12572
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Excitatory Pathways from the Lateral Habenula Enable Propofol-Induced Sedation.
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Curr Biol 2018;28:580
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A Neuronal Hub Binding Sleep Initiation and Body Cooling in Response to a Warm External Stimulus.
Authors: Harding Et al.
Curr Biol 2018;28:2263
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Activity-Dependent Netrin-1 Secretion Drives Synaptic Insertion of GluA1-Containing AMPA Receptors in the Hippocampus.
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Cell Rep 2018;25:168
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The Basolateral Amygdala Is Essential for Rapid Escape: A Human and Rodent Study.
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Cell 2018;175:723
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Astrocytic Activation Generates De Novo Neuronal Potentiation and Memory Enhancement.
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Cell 2018;174:59
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A Neural Circuit for the Suppression of Pain by a Competing Need State.
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Cell 2018;173:140
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Dissociable Structural and Functional Hippocampal Outputs via Distinct Subiculum Cell Classes.
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Cell 2018;173:1280
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Medial Amygdala Kiss1 Neurons Mediate Female Pheromone Stimulation of Luteinizing Hormone in Male Mice.
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Neuroendocrinology 2018;108:172
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The Anterior Insular Cortex?Central Amygdala Glutamatergic Pathway Is Critical to Relapse after Contingency Management.
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Neuron 2017;96:414-427.e8
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Neural Circuit-Specialized Astrocytes: Transcriptomic, Proteomic, Morphological, and Functional Evidence.
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Neuron 2017;95:531
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Loss of Hyperdirect Pathway Cortico-Subthalamic Inputs Following Degeneration of Midbrain Dopamine Neurons.
Authors: Mark D Et al.
Neuron 2017;95:1306-1318.e5
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A Brainstem-Spinal Cord Inhibitory Circuit for Mechanical Pain Modulation by GABA and Enkephalins.
Authors: Francois Et al.
Neuron 2017;93:822
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Glutamatergic projections from the entorhinal cortex to dorsal dentate gyrus mediate context-induced reinstatement of heroin seeking.
Authors: Ge Et al.
Neuropsychopharmacology 2017;42:1860
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Loss of Plasticity in the D2-Accumbens Pallidal Pathway Promotes Cocaine Seeking.
Authors: Heinsbroek Et al.
J Neurosci 2017;37:757
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A tale of two circuits: CCKNTS neuron stimulation controls appetite and induces opposing motivational states by projections to distinct brain regions.
Authors: Roman Et al.
Neuroscience 2017;358:316
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CaMKIIα Expression Defines Two Functionally Distinct Populations of Granule Cells Involved in Different Types of Odor Behavior.
Authors: Martin Et al.
Curr Biol 2017;27:3315-3329.e6
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Cortico-accumbens regulation of approach-avoidance behavior Is modified by experience and chronic pain.
Authors: Schwartz Et al.
Cell Rep 2017;19:1522
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Inhibiting medial septal cholinergic neurons with DREADD alleviated anxiety-like behaviors in mice.
Authors: Zhang Et al.
Neurosci Letts 2017;18:139
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Visuomotor coupling shapes the functional development of mouse visual cortex.
Authors: Attinger
Cell 2017;169(7):1291
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Ethanol-seeking behavior is expressed directly through an extended amygdala to midbrain neural circuit.
Authors: Pina and Cunningham Et al.
Neurobiol Learn Mem 2017;137:83
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Using c-kit to genetically target cerebellar molecular layer interneurons in adult mice.
Authors: Amat
PLoS One 2017;12(6):e0179347
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Orexin-driven GAD65 network of the lateral hypothalamus sets physical activity in mice.
Authors: Cornelia Et al.
Proc Natl Acad Sci U S A 2017;114:4525-4530
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Histamine modulation of the basal ganglia circuitry in the development of pathological grooming.
Authors: Rapanelli
Proc Natl Acad Sci U S A. 2017;114(25):6599
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Appetite controlled by a cholecystokinin nucleus of the solitary tract to hypothalamus neurocircuit.
Authors: D'Agostino Et al.
PLoS Biol 2016;5
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Attenuation of β-Amyloid Deposition and Neurotoxicity by Chemogenetic Modulation of Neural Activity.
Authors: Yuan and Grutzendler
J Neurosci 2016;36:632
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Changes in Female Drosophila Sleep following Mating Are Mediated by SPSN-SAG Neurons.
Authors: Amita Et al.
J Biol Rhythms 2016;31:551-567
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Neural activity promotes long-distance, target-specific regeneration of adult retinal axons.
Authors: Lim Et al.
Nat Neurosci 2016;19:1073
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Warm-Sensitive Neurons that Control Body Temperature.
Authors: Tan Et al.
Cell 2016;167:47
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Ethanol-seeking behavior is expressed directly through an extended amygdala to midbrain neural circuit.
Authors: Pina and Cunningham
Neurobiol Learn Mem 2016;137:83
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DREADDS: Use and application in behavioral neuroscience.
Authors: Smith Et al.
Behav Neurosci 2016;130:137
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CNO was used in pair with DREADD mice for a pain related experiment to determine whether inhibiting PBN neurons will attenuate pain reponses. Publication under second revision for Cell.
Product provides publishable work, no issues arose when dealing with the substance.
I injected clozapine N-oxide in rats (IP, 1 mg/ml/kg). The rats had previously underwent stereotaxic surgery in which a virus carrying a gene for a Gi DREADD was infused unilaterally into the dorsal striatum.
In vivo chemogenetic experiments - CNO was added to the drinking water. While there are issues with the back conversion of CNO to clozapine, this is not the manufacturer's problem. CNO experiments still generate important data as long as they utilize proper controls
Added saccharine to counteract bitter taste of DMSO used to dissolve CNO
We use it as both injectable pass the blood brain barrier ICV, and IP injections. We are looking at both behavioral concern and brain regional efficacy in regard to DREADD. So far, this product has been selectively turning on/off our target cells.
bleed assay
neuron silencer