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61 results for “endocannabinoids”
AP-4-mediated axonal transport controls endocannabinoid production in neurons - Imaging data 3
<p>Imaging data associated with Fig. 6c,e,f, 7e-h, S5a,b, S6b-d and S7 from the article "AP-4-mediated axonal transport controls endocannabinoid production in neurons", published in Nature Communications by Davies et al.</p> <p>DAGLB_Axon_iPSC_derived_neurons: High-throughput confocal imaging was used to assay the density of DAGLB puncta in axons of iPSC neurons from a patient with AP-4 deficiency syndrome (patient 1) and their matched control, labelled with anti-DAGLB, the axonal marker antibody cocktail SMI312 and DAPI.</p> <p>DAGLB_Soma_iPSC_derived_neurons: High-throughput confocal imaging was used to assay the distribution of DAGLB in iPSC-derived neurons from two patients with AP-4 deficiency syndrome (LoF/LoF) and their matched unaffected controls (WT/LoF). Neurons in 96-well plates were labelled with antibodies against DAGLB, GOLGA1 (a TGN marker) and TUJ1, and DAPI. The ratio between the area of high intensity (HI; overlaps with TGN) and low intensity (LI) DAGLB labelling was quantified from three differentiations per cell line.</p> <p>iPSC_derived_neurons_neurite_outgrowth: Neurite outgrowth was assayed in iPSC-derived cortical neurons from two patients with AP4B1-associated AP-4 deficiency syndrome (SPG47) and their unaffected same sex heterozygous parents (control), using automated live cell imaging. Neurons were cultured in the presence of DMSO (vehicle control) or the MGLL inhibitor ABX-1431 at 10, 50, 100 or 500 nM (the highest two doses were administered only to the patient neurons). Neurons were monitored from 4 h post-plating, with images captured every 3 h until 25 h post-plating.</p> <p>See article for methods and further detail.</p>
AP-4-mediated axonal transport controls endocannabinoid production in neurons - Imaging data 2
<p>Imaging data from the article "AP-4-mediated axonal transport controls endocannabinoid production in neurons", published in Nature Communications by Davies et al., from Fig. 5 and S4.</p> <p>Super resolution structured illumination microscopy (SR-SIM) was used to image HeLa cells expressing endogenously-tagged SERINC1-Clover or SERINC3-Clover, treated with siRNA to knock down AP-4 or non-targeting control siRNA, and labelled with anti-GFP and either anti-ATG9A or anti-DAGLB.</p> <p>SR-SIM was performed on a Zeiss Elyra PS.1 microscope equipped with a 63x/1.46 oil objective (alpha Plan-Apochromat 63x/1.46 Oil Korr M27) and a PCO pco.edge 4.2 sCMOS Camera, and controlled with Zeiss ZEN 2 software with the SR-SIM module. Imaging was performed sequentially, with SERINC1 or SERINC3 (labelled with Alexa Fluor Plus 488) imaged first and DAGLB or ATG9A (labelled with Alexa Fluor 568) imaged second, using 3 rotations of the grid pattern and 5 phases for each rotation. Experiments were performed in biological duplicate, from separate dishes of cells, and with immunofluorescence and microscopy performed independently for each replicate. 20 cells were imaged per condition per replicate, and cell selection and manual focus were performed on the first acquired channel only, without viewing the second channel. For channel alignment, slides with multi-coloured fluorescent beads were imaged before and after each experiment using the same acquisition settings. The data were processed using the SIM module in Zen 2 software in manual mode, using default settings and a theoretical PSF model, except for the following settings: Noise filter -1; SR frequency weighting 2; Max.Isotrop on; Baseline shifted on; Raw scale on. Channel alignment fit parameters were calculated in Zen 2 software using a multi-coloured bead image as input, in affine mode. The resulting parameters were tested on further bead images and then applied to the experimental images. The channel-aligned images were used for colocalisation analysis (see methods of article for details).</p> <p>This dataset includes the raw unprocessed data and the processed and channel aligned input data used for the colocalisation analysis.</p> <p>See article for methods and further detail.</p>
AP-4-mediated axonal transport controls endocannabinoid production in neurons - Imaging data 1
<p>Imaging data from the article "AP-4-mediated axonal transport controls endocannabinoid production in neurons", published in Nature Communications by Davies et al., from Fig. 2, 3, 4, 6 and S2.</p> <p>Widefield images were captured on a Leica DMi8 inverted microscope equipped with an iTK LMT200 motorised stage, a 63x/1.47 oil objective (HC PL APO 63x/1.47 OIL) and a Leica DFC9000 GTC Camera, and controlled with LAS X (Leica Application Software X).</p> <p>HeLa_HADAGL_RUSC2_Fig_4: Widefield imaging of wild-type HeLa and HeLa cells stably expressing GFP-RUSC2, transiently expressing HA-DAGLB or HA-DAGLA, and labelled with anti-HA (Alexa Fluor 680), anti-ATG9A (Alexa Fluor 568) and DAPI.</p> <p>HeLa_KD_DAGLB_TGN_Fig_S2a,b: Widefield imaging of immunofluorescence double labelling of DAGLB (Alexa Fluor 568) and TGN46 (Alexa Fluor 680) in HeLa cells transfected with a non-targeting siRNA (control) or with siRNA to knock down AP-4. DAPI labelling of the nucleus is also shown. Note, the antibody used to label AP4E1 (Alexa Fluor 488) in replicate 1 had high non-specific background and so was not used in the analysis.</p> <p>HeLa_KO_DAGLB_TGN_Fig_2a,b: Widefield imaging of immunofluorescence double labelling of DAGLB (Alexa Fluor 568) and TGN46 (Alexa Fluor 680) in wild-type, AP4B1 knockout, and AP4B1 knockout HeLa cells stably expressing AP4B1 (functional rescue). DAPI labelling of the nucleus is also shown.</p> <p>HeLa_RUSC2_DAGLB_Fig_3: Widefield imaging of DAGLB (Alexa Fluor 568) labelling in wild-type and AP-4 knockout (AP4B1 KO or AP4E1 KO) HeLa cells, stably expressing GFP-tagged RUSC2. AP4B1 knockout HeLa expressing RUSC2-GFP were also transiently transfected with AP4B1 (rescue) or mock transfected without DNA as a negative control. DAPI labelling of the nucleus is also shown.</p> <p>iPSC_Neurons_DAGLB_TGN_Fig_6a,b: Widefield imaging of immunofluorescence triple labelling of DAGLB (Alexa Fluor 568), TGN46 (Alexa Fluor 680) and TUJ1 (Alexa Fluor 488; a marker to distinguish neurons from co-cultured astrocytes) in iPSC neurons from a patient with AP-4 deficiency syndrome (patient 1) and their matched control.</p> <p>SHSY5Y_DAGLB_TGN_Fig_2c,d_Fig_S2c,d: Widefield imaging of immunofluorescence double labelling of DAGLB (Alexa Fluor 568) and TGN46 (Alexa Fluor 680) in control (parental Cas9-expressing), AP4B1-depleted (BKO) and AP4E1-depleted (EKO) undifferentiated and neuronally-differentiated SH-SY5Y cells. DAPI labelling of the nucleus is also shown.</p> <p>See article for methods and further detail.</p>
Potentiation of endocannabinoids and other lipid amides prevents hyperalgesia and inflammation in a pre-clinical model of migraine
<p>This dataset comprises the findings obtained in the study aimed at investigating the anti-migraine effects of two URB597 analogs with improved solubility and bioavailability – ARN14633 ([4-fluoro-3-[3-(methylcarbamoyl)phenyl]phenyl] N-cyclohexylcarbamate) and ARN14280 ([3-(3-carbamoylphenyl)-4-(difluoromethoxy)phenyl] N-cyclohexylcarbamate), in the migraine rat model based by nitroglycerin (NTG) administration. Two compounds significantly attenuated the nocifensive behavior induced by NTG compared with original URB597 administered in the same conditions. Additionally, both inhibitors reverted the neurobiological changes induced by NTG in specific peripheral and central areas. Although the precise mechanism underlying these effects remains to be elucidated, our results support further investigational studies of FAAH blockade as a potential therapeutic strategy to treat migraine conditions.</p> <p>ARN14633 (1 mg/kg, i.p.), ARN14280 (3 mg/kg, i.p.) and URB597 (2mg/kg, i.p.) were administered to adult male Sprague-Dawley rats 3 hours after NTG injection. One hour after the administration of either compound, rats were subjected to the orofacial formalin test and compared. In addition, the anti-migraine effects of two URB597 analogs were evaluated by neuronal nitric oxide synthase, pain mediators peptides, pro-inflammatory cytokines gene expression and lipid amides levels in specific areas.</p> <p><strong>The in vivo and ex vivo assessments were:</strong></p> <p>1) mRNA expression levels: CGRP, SP, interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha) and IL-1beta in cervical spinal cord, medulla-pons and trigeminal ganglia. mRNA levels were measured by rt-PCR. All samples were assayed in triplicate and gene expression levels were calculated according to 2−∆∆Ct = 2− (∆Ct gene − ∆Ct housekeeping gene) formula by using Ct (cycle threshold) values.</p> <p>2) Central and peripheral Endocannabinoids levels: Lipid amides levels were measured using a Xevo TQ UPLC-MS/MS system equipped with a reversed-phase BEH C18 column (2.1 × 50 mm, 1.7 μm particle size) (Waters, Milford, USA). The mobile phase consisted of 0.1% formic acid in water as solvent A and 0.1% formic acid in acetonitrile as solvent B. A linear gradient was used: 0.0–0.5 min 20% B; 0.5–2.5 min 20 to 100% B; and 2.5–3.0 min maintained at 100% B. The column was reconditioned to 20% B for 1 min. Analysis time was 4 min and the injection volume was 5 μL.</p> <p><strong>Results in brief</strong></p> <p>ARN14633 and ARN14280 attenuated NTG-induced nocifensive behavior compared with URB597. Additionally, the new FAAH inhibitors reduced transcription of genes encoding neuronal nitric oxide synthase, pain mediators peptides (calcitonin gene-related peptide, substance P) and pro-inflammatory cytokines (tumor necrosis factor-alpha, interleukin-1beta and 6) in the trigeminal ganglion, cervical spinal cord and medulla. Finally, both compounds strongly elevated levels of endocannabinoids and/or other FAAH substrates in cervical spinal cord and medulla, and, to a lesser extent, in the trigeminal ganglia.</p>
Effects of Probiotics on Gut Microbiota, Endocannabinoid and Immune Activation and Symptoms of Fatigue in Dancers
ClinicalTrials.gov study NCT05567653. IPD Sharing: NO. Countries: 1. Publications: 46.
Endocannabinoid signalling in stem cells and cerebral organoids drives differentiation to deep layer projection neurons via CB1 receptors
<p>The endocannabinoid (eCB) system, <i>via</i> cannabinoid CB<sub>1</sub> receptor, regulates neurodevelopment by controlling neural progenitor proliferation and neurogenesis. CB<sub>1</sub> receptor signalling <i>in vivo</i> drives corticofugal deep layer projection neuron development through the regulation of <span>BCL11B </span>and <span>Satb2</span> transcription factors. Here, we investigated the role of eCB signalling in mouse pluripotent embryonic stem cell-derived neuronal differentiation. Characterization of the eCB system revealed increased expression of eCB-metabolizing enzymes, eCB ligands and CB<sub>1</sub> receptors along neuronal differentiation. CB<sub>1</sub> receptor knockdown inhibited neuronal differentiation of deep layer neurons and increased upper layer neuron generation, and this phenotype was rescued by CB<sub>1</sub> re-expression. Pharmacological regulation with CB<sub>1</sub> receptor agonists or elevation of eCB tone with a monoacylglycerol lipase inhibitor promoted neuronal differentiation of deep layer neurons at the expense of upper layer neurons. Patch-clamp analyses revealed that enhancing cannabinoid signalling facilitated neuronal differentiation and functionality. Noteworthy, incubation with CB<sub>1</sub> receptor agonists during human iPSC-derived cerebral organoid formation also promoted the expansion of BCL11B<sup>+</sup> neurons. These findings unveil a cell-autonomous role of eCB signalling that, <i>via</i> CB<sub>1</sub> receptor, promotes mouse and human deep layer cortical neuron development.</p>
Rescue of loss-of-function Long QT Syndrome-associated mutations in Kv7.1/KCNE1 by the endocannabinoid ARA-S
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What's GABA got to do with it? A potential link between the microbiome, schizophrenia, and the endocannabinoid system
<p><span>This study is unique in that it explores specific microbial and metabolic alterations in patients grouped by medications. Further, this study reveals interesting metabolic trends that may be valuable towards unearthing a pathophysiological mechanism for schizophrenia, which is currently lacking. </span></p>
The Kinetics of Endocannabinoids in Patients With Chemotherapy Induced Peripheral Neuropathy by Using Medical Cannabis.
ClinicalTrials.gov study NCT04376437. IPD Sharing: NO. Countries: 1. Publications: 20.
The Endocannabinoid System in Human Gestational Tissues in Labor
ClinicalTrials.gov study NCT03752021. IPD Sharing: Not stated. Countries: 1. Publications: 20.
Gene Expression of Endocannabinoid System in Episodic and Chronic Migraine Patients
ClinicalTrials.gov study NCT04324710. IPD Sharing: Not stated. Countries: 1. Publications: 12.
Gastric Bypass and Peripheral Activity of the Endocannabinoid System
ClinicalTrials.gov study NCT01003873. IPD Sharing: Not stated. Countries: 1. Publications: 5.
Endocannabinoid Activity Remodulation for Psychosis Liability in Youth
ClinicalTrials.gov study NCT06037993. IPD Sharing: Not stated. Countries: 1. Publications: 1.
Stress and Opioid Misuse Risk: The Role of Endogenous Opioid and Endocannabinoid Mechanisms
ClinicalTrials.gov study NCT05142267. IPD Sharing: NO. Countries: 1. Publications: 42.
The Role of Endocannabinoids in Insulin Production and Action
ClinicalTrials.gov study NCT01517100. IPD Sharing: UNDECIDED. Countries: 1. Publications: 3.
Endocannabinoid signalling in stem cells and cerebral organoids drives differentiation to deep layer projection neurons via CB1 receptors
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Supplementary table 1. Dunn's test results of the endocannabinoids level analyses.
<p>A supplementary table to manuscript entitled 'Co-administration of a cannabinoid receptor 1 antagonist, AM6545 and a nociceptin receptor agonist, SCH221510 increases the severity of DSS-induced colitis in mice'. The table presents the results of Dunn's post-hoc test for multiple comparisons, performed following the Kruskal-Wallis test, to analyze group differences in endocannabinoid level analyses.</p>
Endocannabinoid and Psychological Responses to Acute Resistance Exercise in Trained and Untrained Adults
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Oral Endocannabinoids in People With Prediabetes and Diabetes
ClinicalTrials.gov study NCT06040164. IPD Sharing: Not stated. Countries: 0. Publications: 24.
Effect of Cannabis and Endocannabinoids on HIV Neuropathic Pain
ClinicalTrials.gov study NCT03099005. IPD Sharing: YES. Countries: 1. Publications: 0.
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Allen Brain Atlas
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