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  1. Article ; Online: Western Blotting of the Endocannabinoid System.

    Wager-Miller, Jim / Mackie, Ken

    Methods in molecular biology (Clifton, N.J.)

    2022  Volume 2576, Page(s) 385–393

    Abstract: Measuring protein levels of receptors and enzymes involved in endocannabinoid metabolism is an important step for understanding the distribution, function, and regulation of these components of the endocannabinoid system. A common approach for detecting ... ...

    Abstract Measuring protein levels of receptors and enzymes involved in endocannabinoid metabolism is an important step for understanding the distribution, function, and regulation of these components of the endocannabinoid system. A common approach for detecting proteins from complex biological systems is western blotting. In this chapter, we describe a general approach to western blotting protein components of the endocannabinoid system using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and nitrocellulose membranes with a focus on detecting type 1 cannabinoid (CB
    MeSH term(s) Blotting, Western ; Cannabinoids ; Collodion ; Endocannabinoids/metabolism ; Sodium Dodecyl Sulfate
    Chemical Substances Cannabinoids ; Endocannabinoids ; Sodium Dodecyl Sulfate (368GB5141J) ; Collodion (9004-70-0)
    Language English
    Publishing date 2022-09-24
    Publishing country United States
    Document type Journal Article
    ISSN 1940-6029
    ISSN (online) 1940-6029
    DOI 10.1007/978-1-0716-2728-0_32
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  2. Article ; Online: Quantitation of Plasma Membrane (G Protein-Coupled) Receptor Trafficking in Cultured Cells.

    Wager-Miller, Jim / Mackie, Ken

    Methods in molecular biology (Clifton, N.J.)

    2022  Volume 2576, Page(s) 395–406

    Abstract: Measuring the functional behavior of G protein-coupled receptors (GPCRs) has been a major focus of academic and pharmaceutical research for many decades. These efforts have led to the development of many assays to measure the downstream effects of ligand ...

    Abstract Measuring the functional behavior of G protein-coupled receptors (GPCRs) has been a major focus of academic and pharmaceutical research for many decades. These efforts have led to the development of many assays to measure the downstream effects of ligand binding on receptor activity. In this chapter, we describe an internalization/recycling assay that can be used to track changes in receptor number at the plasma membrane. Used in concert with other assays, this antibody-based technique can provide dynamic information on GPCR activation by receptor-specific ligands.
    MeSH term(s) Cell Membrane/metabolism ; Cells, Cultured ; GTP-Binding Proteins/metabolism ; Ligands ; Receptors, G-Protein-Coupled/metabolism
    Chemical Substances Ligands ; Receptors, G-Protein-Coupled ; GTP-Binding Proteins (EC 3.6.1.-)
    Language English
    Publishing date 2022-09-24
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural
    ISSN 1940-6029
    ISSN (online) 1940-6029
    DOI 10.1007/978-1-0716-2728-0_33
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  3. Article ; Online: Cannabinoid Receptor Type 1 Activation Causes a Water Diuresis by Inducing an Acute Central Diabetes Insipidus in Mice.

    Rein, Joshua L / Mackie, Ken / Kleyman, Thomas R / Satlin, Lisa M

    American journal of physiology. Renal physiology

    2024  

    Abstract: Cannabis and synthetic cannabinoid consumption is increasing worldwide. Cannabis contains numerous phytocannabinoids that act on the G-protein-coupled cannabinoid receptors type 1 (CB1R) and type 2 (CB2R) expressed throughout the body, including the ... ...

    Abstract Cannabis and synthetic cannabinoid consumption is increasing worldwide. Cannabis contains numerous phytocannabinoids that act on the G-protein-coupled cannabinoid receptors type 1 (CB1R) and type 2 (CB2R) expressed throughout the body, including the kidney. Essentially every organ, including the kidney, produces endocannabinoids (ECs), endogenous ligands to these receptors. Cannabinoids acutely increase urine output in rodents and humans, thus potentially influencing total-body water and electrolyte homeostasis. As the kidney collecting duct (CD) regulates total body water, acid/base, and electrolyte balance through specific functions of principal cells (PCs) and intercalated cells (ICs), we examined the cell-specific immunolocalization of CB1R in the mouse CD. Antibodies against either the C-terminus or N-terminus of CB1R consistently labeled AQP2(-) cells in the cortical and medullary CD, and thus presumably ICs. Given the well-established role of ICs in urinary acidification, we utilized a clearance approach in mice that were acid-loaded with 280 mM NH
    Language English
    Publishing date 2024-04-18
    Publishing country United States
    Document type Journal Article
    ZDB-ID 603837-2
    ISSN 1522-1466 ; 0363-6127
    ISSN (online) 1522-1466
    ISSN 0363-6127
    DOI 10.1152/ajprenal.00320.2022
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  4. Article: SGIP1 in axons prevents internalization of desensitized CB1R and modifies its function.

    Durydivka, Oleh / Mackie, Ken / Blahos, Jaroslav

    Frontiers in neuroscience

    2023  Volume 17, Page(s) 1213094

    Abstract: In the central nervous system (CNS), cannabinoid receptor 1 (CB1R) is preferentially expressed in axons where it has a unique property, namely resistance to agonist-driven endocytosis. This review aims to summarize what we know about molecular mechanisms ...

    Abstract In the central nervous system (CNS), cannabinoid receptor 1 (CB1R) is preferentially expressed in axons where it has a unique property, namely resistance to agonist-driven endocytosis. This review aims to summarize what we know about molecular mechanisms of CB1R cell surface stability in axonal compartments, how these impact CB1R signaling, and to consider their physiological consequences. This review then focuses on a potential candidate for maintaining axonal CB1R at the cell surface, Src homology 3-domain growth factor receptor-bound 2-like endophilin interacting protein 1 (SGIP1). SGIP1 may contribute to the polarized distribution of CB1R and modify its signaling in axons. In addition, deletion of SGIP1 results in discrete behavioral changes in modalities controlled by the endocannabinoid system
    Language English
    Publishing date 2023-07-20
    Publishing country Switzerland
    Document type Journal Article ; Review
    ZDB-ID 2411902-7
    ISSN 1662-453X ; 1662-4548
    ISSN (online) 1662-453X
    ISSN 1662-4548
    DOI 10.3389/fnins.2023.1213094
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  5. Article ; Online: Hexahydrocannabinol (HHC) and Δ

    Durydivka, Oleh / Palivec, Petr / Gazdarica, Matej / Mackie, Ken / Blahos, Jaroslav / Kuchar, Martin

    Scientific reports

    2024  Volume 14, Issue 1, Page(s) 9181

    Abstract: The Cannabis sativa plant has been used for centuries as a recreational drug and more recently in the treatment of patients with neurological or psychiatric disorders. In many instances, treatment goals include relief from posttraumatic disorders, ... ...

    Abstract The Cannabis sativa plant has been used for centuries as a recreational drug and more recently in the treatment of patients with neurological or psychiatric disorders. In many instances, treatment goals include relief from posttraumatic disorders, anxiety, or to support treatment of chronic pain. Ligands acting on cannabinoid receptor 1 (CB1R) are also potential targets for the treatment of other health conditions. Using an evidence-based approach, pharmacological investigation of CB1R agonists is timely, with the aim to provide chronically ill patients relief using well-defined and characterized compounds from cannabis. Hexahydrocannabinol (HHC), currently available over the counter in many countries to adults and even children, is of great interests to policy makers, legal administrators, and healthcare regulators, as well as pharmacologists. Herein, we studied the pharmacodynamics of HHC epimers, which activate CB1R. We compared their key CB1R-mediated signaling pathway activities and compared them to the pathways activated by Δ
    MeSH term(s) Dronabinol/pharmacology ; Receptor, Cannabinoid, CB1/metabolism ; Receptor, Cannabinoid, CB1/agonists ; Signal Transduction/drug effects ; Humans ; Cannabinol/pharmacology ; Animals ; Cannabinoid Receptor Agonists/pharmacology ; HEK293 Cells ; Mice
    Chemical Substances Dronabinol (7J8897W37S) ; Receptor, Cannabinoid, CB1 ; Cannabinol (7UYP6MC9GH) ; Cannabinoid Receptor Agonists
    Language English
    Publishing date 2024-04-22
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, N.I.H., Extramural
    ZDB-ID 2615211-3
    ISSN 2045-2322 ; 2045-2322
    ISSN (online) 2045-2322
    ISSN 2045-2322
    DOI 10.1038/s41598-024-58845-7
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  6. Article ; Online: Alternative pain management via endocannabinoids in the time of the opioid epidemic: Peripheral neuromodulation and pharmacological interventions.

    Lee, Ming Tatt / Mackie, Ken / Chiou, Lih-Chu

    British journal of pharmacology

    2022  Volume 180, Issue 7, Page(s) 894–909

    Abstract: The use of opioids in pain management is hampered by the emergence of analgesic tolerance, which leads to increased dosing and side effects, both of which have contributed to the opioid epidemic. One promising potential approach to limit opioid analgesic ...

    Abstract The use of opioids in pain management is hampered by the emergence of analgesic tolerance, which leads to increased dosing and side effects, both of which have contributed to the opioid epidemic. One promising potential approach to limit opioid analgesic tolerance is activating the endocannabinoid system in the CNS, via activation of CB
    MeSH term(s) Humans ; Endocannabinoids/metabolism ; Analgesics, Opioid/adverse effects ; Pain Management ; Opioid Epidemic ; Drug Tolerance ; Pain/drug therapy ; Pain/metabolism ; Analgesics/pharmacology ; Receptor, Cannabinoid, CB1
    Chemical Substances Endocannabinoids ; Analgesics, Opioid ; Analgesics ; Receptor, Cannabinoid, CB1
    Language English
    Publishing date 2022-01-21
    Publishing country England
    Document type Journal Article ; Review ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 80081-8
    ISSN 1476-5381 ; 0007-1188
    ISSN (online) 1476-5381
    ISSN 0007-1188
    DOI 10.1111/bph.15771
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  7. Article: Adolescent Δ

    Chen, Han-Ting / Mackie, Ken

    Frontiers in psychiatry

    2020  Volume 11, Page(s) 576214

    Abstract: As the frequency of cannabis use by 14-16-year-olds increases, it becomes increasingly important to understand the effect of cannabis on the developing central nervous system. Using mice as a model system, we treated adolescent (28 day old) C57BL6/J mice ...

    Abstract As the frequency of cannabis use by 14-16-year-olds increases, it becomes increasingly important to understand the effect of cannabis on the developing central nervous system. Using mice as a model system, we treated adolescent (28 day old) C57BL6/J mice of both sexes for 3 weeks with 3 mg/kg tetrahydrocannabinol (THC). Starting a week after the last treatment, several cognitive behaviors were analyzed. Mice treated with THC as adolescents acquired proficiency in a working memory task more slowly than vehicle-treated mice. Working memory recall in both sexes of THC-treated mice was also deficient during increasing cognitive load compared to vehicle-treated mice. Our adolescent THC treatment did not strongly affect social preference, anxiety behaviors, or decision-making behaviors on the elevated T maze task. In summary, under the conditions of this study, adolescent THC treatment of mice markedly affected the establishment, and persistence of working memory, while having little effect on decision-making, social preference or anxiety behaviors. This study provides further support that adolescent THC affects specific behavioral domains.
    Language English
    Publishing date 2020-11-12
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2564218-2
    ISSN 1664-0640
    ISSN 1664-0640
    DOI 10.3389/fpsyt.2020.576214
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  8. Article ; Online: Review of the Endocannabinoid System.

    Lu, Hui-Chen / Mackie, Ken

    Biological psychiatry. Cognitive neuroscience and neuroimaging

    2020  Volume 6, Issue 6, Page(s) 607–615

    Abstract: The endocannabinoid system (ECS) is a widespread neuromodulatory network involved in the developing central nervous system as well as playing a major role in tuning many cognitive and physiological processes. The ECS is composed of endogenous ... ...

    Abstract The endocannabinoid system (ECS) is a widespread neuromodulatory network involved in the developing central nervous system as well as playing a major role in tuning many cognitive and physiological processes. The ECS is composed of endogenous cannabinoids, cannabinoid receptors, and the enzymes responsible for the synthesis and degradation of endocannabinoids. In addition to its endogenous roles, cannabinoid receptors are the primary target of Δ
    MeSH term(s) Cannabis ; Endocannabinoids ; Humans ; Neuronal Plasticity ; Receptors, Cannabinoid
    Chemical Substances Endocannabinoids ; Receptors, Cannabinoid
    Language English
    Publishing date 2020-08-01
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Review
    ZDB-ID 2879089-3
    ISSN 2451-9030 ; 2451-9022
    ISSN (online) 2451-9030
    ISSN 2451-9022
    DOI 10.1016/j.bpsc.2020.07.016
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  9. Article ; Online: Prefrontal cortical distribution of muscarinic M2 and cannabinoid-1 (CB1) receptors in adult male mice with or without chronic adolescent exposure to Δ9-tetrahydrocannabinol.

    Garzón, Miguel / Chan, June / Mackie, Ken / Pickel, Virginia M

    Cerebral cortex (New York, N.Y. : 1991)

    2022  Volume 32, Issue 23, Page(s) 5420–5437

    Abstract: Chronic adolescent administration of marijuana's major psychoactive compound, ∆9-tetrahydrocannabinol (Δ9-THC), produces adaptive changes in adult social and cognitive functions sustained by prelimbic prefrontal cortex (PL-PFC). Memory and learning ... ...

    Abstract Chronic adolescent administration of marijuana's major psychoactive compound, ∆9-tetrahydrocannabinol (Δ9-THC), produces adaptive changes in adult social and cognitive functions sustained by prelimbic prefrontal cortex (PL-PFC). Memory and learning processes in PL-PFC neurons can be regulated through cholinergic muscarinic-2 receptors (M2R) and modulated by activation of cannabinoid-1 receptors (CB1Rs) targeted by Δ9-THC. Thus, chronic exposure to Δ9-THC during adolescence may alter the expression and/or distribution of M2Rs in PL-PFC neurons receiving CB1R terminals. We tested this hypothesis by using electron microscopic dual CB1R and M2R immunolabeling in adult C57BL/6 J male mice that had received vehicle or escalating dose of Δ9-THC through adolescence. In vehicle controls, CB1R immunolabeling was mainly localized to axonal profiles virtually devoid of M2R but often apposing M2R-immunoreactive dendrites and dendritic spines. The dendrites received inputs from CB1R-labeled or unlabeled terminals, whereas spines received asymmetric synapses exclusively from axon terminals lacking CB1Rs. Adolescent Δ9-THC significantly increased plasmalemmal M2R-immunogold density exclusively in large dendrites receiving input from CB1R-labeled terminals. In contrast, cytoplasmic M2R-immunogold density decreased in small spines of the Δ9-THC-treated adult mice. We conclude that Δ9-THC engagement of CB1Rs during adolescence increases M2R plasmalemmal accumulation in large proximal dendrites and decreases M2R cytoplasmic expression in small spines of PL-PFC.
    MeSH term(s) Animals ; Male ; Mice ; Dronabinol/pharmacology ; Mice, Inbred C57BL ; Prefrontal Cortex/metabolism ; Receptor, Cannabinoid, CB1/metabolism ; Receptor, Muscarinic M2/metabolism
    Chemical Substances Dronabinol (7J8897W37S) ; Receptor, Cannabinoid, CB1 ; Receptor, Muscarinic M2
    Language English
    Publishing date 2022-02-11
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural
    ZDB-ID 1077450-6
    ISSN 1460-2199 ; 1047-3211
    ISSN (online) 1460-2199
    ISSN 1047-3211
    DOI 10.1093/cercor/bhac024
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    In stock of ZB MED Cologne/Königswinter

    Zs.A 3235: Show issues Location:
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  10. Article ; Online: A collection of cannabinoid-related negative findings from autaptic hippocampal neurons.

    Straiker, Alex / Dvorakova, Michaela / Bosquez-Berger, Taryn / Blahos, Jaroslav / Mackie, Ken

    Scientific reports

    2023  Volume 13, Issue 1, Page(s) 9610

    Abstract: Autaptic hippocampal neurons are an architecturally simple model of neurotransmission that express several forms of cannabinoid signaling. Over the past twenty years this model has proven valuable for studies ranging from enzymatic control of ... ...

    Abstract Autaptic hippocampal neurons are an architecturally simple model of neurotransmission that express several forms of cannabinoid signaling. Over the past twenty years this model has proven valuable for studies ranging from enzymatic control of endocannabinoid production and breakdown, to CB
    MeSH term(s) Cannabinoids/pharmacology ; Neurons ; Endocannabinoids ; Synaptic Transmission ; Hippocampus
    Chemical Substances Cannabinoids ; Endocannabinoids
    Language English
    Publishing date 2023-06-13
    Publishing country England
    Document type Journal Article
    ZDB-ID 2615211-3
    ISSN 2045-2322 ; 2045-2322
    ISSN (online) 2045-2322
    ISSN 2045-2322
    DOI 10.1038/s41598-023-36710-3
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