Molecular networks underlying cannabinoid signaling in skeletal muscle plasticity

Journal of Cellular Physiology, Journal Year: 2022, Volume and Issue: 237(9), P. 3517 - 3540

Published: July 21, 2022

Abstract The cannabinoid system is ubiquitously present and classically considered to engage in neural immunity processes. Yet, the role of whole body tissue metabolism via central peripheral mechanisms increasingly recognized. review provides insights (i) how signaling regulated receptor‐independent ‐dependent (ii) these cascades (might) affect skeletal muscle plasticity physiology. Receptor‐independent include endocannabinoid eicosanoids regulation ion channels. Alternatively, endocannabinoids can act as ligands for different classic (cannabinoid receptor 1 [CB ], CB 2 ) and/or alternative (e.g., TRPV1, GPR55) receptors with a unique affinity, specificity, intracellular cascade (often tissue‐specific). Antagonism might hold clues improve oxidative (mitochondrial) metabolism, insulin sensitivity, satellite cell growth, anabolism, whereas agonism be promising way stimulate growth. Besides, ameliorates regeneration macrophage polarization toward an anti‐inflammatory phenotype, induction MyoD myogenin expression antifibrotic mechanisms. Also TRPV1 GPR55 contribute growth metabolism. Future studies should reveal targeted quantity quality conditions such ageing, disease, disuse, metabolic dysregulation, taking into account challenges that are inherent modulation system, side effects.

Language: Английский

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High-throughput combined voltage-clamp/current-clamp analysis of freshly isolated neurons

Cell Reports Methods, Journal Year: 2023, Volume and Issue: 3(1), P. 100385 - 100385

Published: Jan. 1, 2023

The patch-clamp technique is the gold-standard methodology for analysis of excitable cells. However, throughput manual slow, and high-throughput robotic patch-clamp, while helpful applications like drug screening, has been primarily used to study channels receptors expressed in heterologous systems. We introduce an approach automated patch-clamping that enhances cells at channel cellular levels. This involves dissociating isolating neurons from intact tissues using a instrument, followed by open-source Python script filtration. As proof concept, we apply this investigate biophysical properties voltage-gated sodium (Nav) dorsal root ganglion (DRG) neurons, which are among most diverse complex neuronal Our enables voltage- current-clamp recordings same cell, allowing unbiased, fast, simultaneous, head-to-head electrophysiological wide range freshly isolated without requiring culturing on coverslips.

Language: Английский

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Cannabidiol potentiates hyperpolarization-activated cyclic nucleotide–gated (HCN4) channels

The Journal of General Physiology, Journal Year: 2024, Volume and Issue: 156(6)

Published: April 23, 2024

Cannabidiol (CBD), the main non-psychotropic phytocannabinoid produced by Cannabis sativa plant, blocks a variety of cardiac ion channels. We aimed to identify whether CBD regulated pacemaker channel or hyperpolarization-activated cyclic nucleotide–gated (HCN4). HCN4 channels are important for generation action potential in sinoatrial node heart and increased rate response β-adrenergic stimulation. were expressed HEK 293T cells, effect application was examined using whole-cell patch clamp. found that depolarized V1/2 activation holo-HCN4 channels, with an EC50 1.6 µM, without changing current density. also sped kinetics approximately threefold. potentiation occurred via binding closed state channel. CBD’s mechanism distinct from cAMP, as potentiated apo-HCN4 The addition exogenous PIP2 analog did not alter ability potentiate suggesting acts unique known potentiator PIP2. Lastly, gain insight into action, computational modeling targeted mutagenesis used predict binds lipid-binding pocket at C-terminus voltage sensor. represents first FDA-approved drug our findings suggest novel starting point development targeting

Language: Английский

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Unplugging lateral fenestrations of NALCN reveals a hidden drug binding site within the pore region

Proceedings of the National Academy of Sciences, Journal Year: 2024, Volume and Issue: 121(22)

Published: May 24, 2024

The sodium (Na + ) leak channel (NALCN) is a member of the four-domain voltage-gated cation family that includes prototypical and calcium channels V s Ca s, respectively). Unlike Na which have four lateral fenestrations serve as routes for lipophilic compounds to enter central cavity modulate function, NALCN has bulky residues (W311, L588, M1145, Y1436) block these openings. Structural data suggest occluded underlie pharmacological resistance NALCN, but functional evidence lacking. To test this hypothesis, we unplugged by substituting aforementioned with alanine (AAAA) compared effects , blockers on both wild-type (WT) AAAA channels. Most behaved in similar manner channels, phenytoin 2-aminoethoxydiphenyl borate (2-APB) elicited additional, distinct responses Further experiments using single mutants revealed 2-APB inner through fenestrations, implying structural specificity their modes access. Using combination computational approaches, identified amino acid critical activity, supporting existence drug binding site(s) within pore region. Intrigued activity its analogues, tested containing diphenylmethane/amine moiety WT We clinically used drugs exhibited diverse thus expanding toolbox NALCN. While low potencies active reiterate our findings lay foundation rational design develop modulators refined properties.

Language: Английский

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Characterizing fenestration size in sodium channel subtypes and their accessibility to inhibitors

Biophysical Journal, Journal Year: 2021, Volume and Issue: 121(2), P. 193 - 206

Published: Dec. 24, 2021

Language: Английский

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Late sodium current: incomplete inactivation triggers seizures, myotonias, arrhythmias, and pain syndromes

The Journal of Physiology, Journal Year: 2022, Volume and Issue: 600(12), P. 2835 - 2851

Published: April 18, 2022

Acquired and inherited dysfunction in voltage-gated sodium channels underlies a wide range of diseases. In addition to defects trafficking expression, channelopathies are caused by one or several gating properties, for instance activation inactivation. Disruption channel inactivation leads increased late current, which is common defect seizure disorders, cardiac arrhythmias skeletal muscle myotonia pain. An increase current repetitive action potentials neurons muscles, prolonged potential duration the heart. this Topical Review, we compare effects brain, heart, peripheral nerves.

Language: Английский

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Anti‐inflammatory effects of cannabidiol against lipopolysaccharides in cardiac sodium channels

British Journal of Pharmacology, Journal Year: 2022, Volume and Issue: 179(24), P. 5259 - 5272

Published: July 30, 2022

Sepsis, caused by a dysregulated response to infections, can lead cardiac arrhythmias. However, the mechanisms underlying sepsis-induced inflammation, and how inflammation provokes arrhythmias, are not well understood. We hypothesized that cannabidiol (CBD) may ameliorate lipopolysaccharide (LPS)-induced cardiotoxicity, via Toll-like receptors (TLR4) sodium channels (NaV 1.5).We incubated human immune cells (THP-1 macrophages) with LPS for 24 h, then extracted THP-1 incubation media. ELISA assays showed (1 or 5 μg·ml-1 ), in concentration-dependent manner, MPLA (TLR4 agonist, ) stimulated release inflammatory cytokines (TNF-α IL-6). Prior (4 h) CBD (5 μM) C34 antagonist: inhibited MPLA-induced of both IL-6 TNF-α. Human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) were subsequently h media from LPS, alone, combination C34. Voltage-clamp experiments right shift voltage dependence NaV 1.5 activation, steady state fast inactivation (SSFI), increased persistent current prolonged silico action potential duration hiSPC-CMs MPLA-THP-1 Co-incubation rescued biophysical dysfunction MPLA.Our results suggest protect against subsequent arrhythmias through (i) inhibition cytokines, antioxidant anti-apoptotic effects and/or (ii) direct effect on 1.5.

Language: Английский

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Transient receptor potential vanilloid 1 (TRPV1)‐independent actions of capsaicin on cellular excitability and ion transport

Medicinal Research Reviews, Journal Year: 2023, Volume and Issue: 43(4), P. 1038 - 1067

Published: March 14, 2023

Abstract Capsaicin is a naturally occurring alkaloid derived from chili pepper that responsible for its hot pungent taste. known to exert multiple pharmacological actions, including analgesia, anticancer, anti‐inflammatory, antiobesity, and antioxidant effects. The transient receptor potential vanilloid subfamily member 1 (TRPV1) the main mediating majority of capsaicin However, numerous studies suggest TRPV1 not only target capsaicin. An increasing number indicates capsaicin, at low mid µM ranges, indirectly through TRPV1‐mediated Ca 2+ increases, but also directly modulates functions voltage‐gated Na + , K channels, as well ligand‐gated ion channels other transporters enzymes involved in cellular excitability. These TRPV1‐independent effects are mediated by alterations biophysical properties lipid membrane subsequent modulation functional direct binding channels. present study, first time, systematically categorizes this diverse range non‐TRPV1 targets discusses molecular mechanisms excitable, nonexcitable cells.

Language: Английский

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Functionally-selective inhibition of threshold sodium currents and excitability in dorsal root ganglion neurons by cannabinol

Communications Biology, Journal Year: 2024, Volume and Issue: 7(1)

Published: Jan. 23, 2024

Abstract Cannabinol (CBN), an incompletely understood metabolite for ∆9-tetrahydrocannabinol, has been suggested as analgesic. CBN interacts with endocannabinoid (CB) receptors, but is also reported to interact non-CB targets, including various ion channels. We assessed effects on voltage-dependent sodium (Nav) channels expressed heterologously and in native dorsal root ganglion (DRG) neurons. Our results indicate that a functionally-selective, structurally-non-selective Nav current inhibitor. CBN’s main effect slow inactivation. slows recovery from slow-inactivated states, hyperpolarizes steady-state inactivation, enter deeper slower inactivated states. Multielectrode array recordings attenuates DRG neuron excitability. Voltage- current-clamp analysis of freshly isolated neurons via our automated patch-clamp platform confirmed these findings. The inhibitory currents excitability add new dimension its actions suggest this cannabinoid may be useful neuropathic pain.

Language: Английский

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In Vitro Inhibition of Voltage-Dependent Sodium Currents by the Antifungal Drug Amorolfine

Journal of Biological Chemistry, Journal Year: 2025, Volume and Issue: unknown, P. 108407 - 108407

Published: March 1, 2025

Voltage-gated sodium (Nav) channels are critical for electrical signaling, and their pharmacological modulation can be leveraged the development of therapeutic agents targeting various disorders. The local anesthetic (LA) site on Nav is particularly important, as it a common target many clinically used inhibitors, including anticonvulsants antiarrhythmics. Our goal was to identify novel channel inhibitors by leveraging physicochemical criteria, focusing potential LA binding candidates. We identified amorolfine (AMF), phenyl-propyl morpholine derivative, putative modulator. results demonstrate that AMF acts state-dependent inhibitor channels, with ∼30-fold preference inactivated states. It stabilizes inactivation prevents from conducting, driven through its stabilization inactivation. These findings suggest represents new compound inhibits offering insights into future potentially other ion channels.

Language: Английский

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