Functional characterization of 5-HT1A and 5-HT1B serotonin receptor signaling through G-protein-activated inwardly rectifying K+ channels in a fluorescence-based membrane potential assay

Gadgaard, Camilla; Jensen, Anders A.

doi:10.1016/j.bcp.2020.113870

Cited by 7 publications

(10 citation statements)

References 72 publications

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“…Em changes were analyzed by fluorescencebased plate reader (FLIPR) Membrane Potential Assay (Molecular Devices) (15)(16)(17). Relative fluorescence of Em red dye-loaded cells was recorded every 7 seconds using a Synergy-2 Multi-Mode Microplate Reader (Biotek; excitation 525 nm, emission 590 nm, at 37 8 C) after adding LPS (2 mg/ml; Cell Signaling; E. coli serotype O111:B4), NS1619 (30 mM; Millipore), NS19504 (30 mM; Alomone Labs), Paxilline (10 mM; Alomone Labs), or equimolar vehicle controls.…”

Section: Em Measurementsmentioning

confidence: 99%

BK Channels Regulate LPS-induced CCL-2 Release from Human Pulmonary Endothelial Cells

Zyrianova¹,

López²,

Liao³

et al. 2021

Am J Respir Cell Mol Biol

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We recently established a role for the stretch-activated two-poredomain K 1 (K2P) channel TREK-1 (K2P2.1) in inflammatory cytokine secretion using models of hyperoxia-, mechanical stretch-, and TNF-a-induced acute lung injury. We have now discovered the expression of large conductance, Ca 21 -activated K 1 (BK) channels in human pulmonary microvascular endothelial cells and primary human alveolar epithelial cells using semiquantitative real-time PCR, IP and Western blot, and investigated their role in inflammatory cytokine secretion using an LPS-induced acute lung injury model. As expected, LPS induced IL-6 and CCL-2 secretion from pulmonary endothelial and epithelial cells. BK activation with NS1619 decreased LPS-induced CCL-2 but not IL-6 secretion from endothelial cells and had no effect on epithelial cells, although fluorometric assays revealed that BK activation hyperpolarized the plasma membrane potential (Em) of both cell types. Interestingly, BK inhibition (Paxilline) did not alter cytokine secretion or the Em in either cell type. Furthermore, LPS treatment by itself did not affect the Em or intracellular Ca 21 concentrations. Therefore, we propose BK channel activation as a novel targeted approach to counteract LPS-induced CCL-2 secretion from endothelial cells. This protective effect appears to occur via Em hyperpolarization but independent of intracellular Ca 21 concentrations.

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Section: Em Measurementsmentioning

confidence: 99%

BK Channels Regulate LPS-induced CCL-2 Release from Human Pulmonary Endothelial Cells

Zyrianova¹,

López²,

Liao³

et al. 2021

Am J Respir Cell Mol Biol

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“…The induced and quiescent stable cell lines were stimulated for 5 min by graded concentrations of EG (10 −8 and 10 −7 M), and the selective agonis at 10 −7 M, that were DOI, dopamine, ramelteon and OxB, respectively corresponding to 5HT 2A , D 2 , MT 2 and OX 2 cells, using FBS as positive control and deionized water as negative control. EG: emodin-8-O-β- d -glucopyranoside; FBS: fetal bovine serum; ERK: extracellular regulated protein kinases; CFP: cyan fluorescent protein; 5-HT 1B : serotonin receptor 1B; 5-HT 2A : serotonin receptor 2A; D 2 : dopamine receptor 2; MT 2 : melatonin receptor 2; OX 2 : orexin receptor 2; VSV: vesicular stomatitis virus; mGluR5: metabotropic glutamate receptor 5; FBS: fetal bovine serum; DOI (1-(4-iodo-2,5-dimethoxyphenyl) propan-2-amine; dopamine: 4-(2-Aminoethyl) benzene-1,2-diol; ramelteon (N-[2-[(8S)-2.6,7,8-tetrahydro-1H-cyclopenta [e] ( Gadgaard and Jensen, 2020 ) benzoxol-8-yl]ethyl]propanamide).…”

Section: Resultsmentioning

confidence: 99%

“…The serotonin receptor 5-HT 1B , is an important part of the serotonergic system, which regulates crucial processes of the CNS such as cognition, satiety, anxiety, depression, and sleep. 5-HT 1B has been considered a drug target for serotonergic system-related disorders ( Gadgaard and Jensen, 2020 ; Monti and Jantos, 2008 ; Tiger et al, 2018 ). We identified 5-HT 1B as a target of EG, an active ingredient in several Chinese medicinal plants with antidepressant and neuroprotective effects, by using molecular docking, DARTS, and CETSA.…”

Section: Discussionmentioning

confidence: 99%

“…These effects are mediated by serotonin receptors, such as the serotonin receptor subtype 1B (5-HT 1B ). These receptors, belonging to the G protein-coupled receptors (GPCRs) superfamily, are abundantly expressed in the CNS and constitute validated as well as putative drug targets in a variety of cognitive and psychiatric disorders, including depression and Alzheimer’s disease (AD), the most common incurable neurodegenerative disease ( Monti and Jantos, 2008 ; Tiger et al, 2018 ; Gadgaard and Jensen, 2020 ).…”

Section: Introductionmentioning

confidence: 99%

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A Novel 5-HT1B Receptor Agonist of Herbal Compounds and One of the Therapeutic Uses for Alzheimer’s Disease

Yang

Zhang

et al. 2021

Front. Pharmacol.

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The serotonin receptor 5-HT1B is widely expressed in the central nervous system and has been considered a drug target in a variety of cognitive and psychiatric disorders. The anti-inflammatory effects of 5-HT1B agonists may present a promising approach for Alzheimer’s disease (AD) treatment. Herbal antidepressants used in the treatment of AD have shown functional overlap between the active compounds and 5-HT1B receptor stimulation. Therefore, compounds in these medicinal plants that target and stimulate 5-HT1B deserve careful study. Molecular docking, drug affinity responsive target stability, cellular thermal shift assay, fluorescence resonance energy transfer (FRET), and extracellular regulated protein kinases (ERK) 1/2 phosphorylation tests were used to identify emodin-8-O-β-d-glucopyranoside (EG), a compound from Chinese medicinal plants with cognitive deficit attenuating and antidepressant effects, as an agonist of 5-HT1B. EG selectively targeted 5-HT1B and activated the 5-HT1B-induced signaling pathway. The activated 5-HT1B pathway suppressed tumor necrosis factor (TNF)-α levels, thereby protecting neural cells against beta-amyloid (Aβ)-induced death. Moreover, the agonist activity of EG towards 5-HT1B receptor, in FRET and ERK1/2 phosphorylation, was antagonized by SB 224289, a 5-HT1B antagonist. In addition, EG relieved AD symptoms in transgenic worm models. These results suggested that 5-HT1B receptor activation by EG positively affected Aβ-related inflammatory process regulation and neural death resistance, which were reversed by antagonist SB 224289. The active compounds such as EG might act as potential therapeutic agents through targeting and stimulating 5-HT1B receptor for AD and other serotonin-related disorders. This study describes methods for identification of 5-HT1B agonists from herbal compounds and for evaluating agonists with biological functions, providing preliminary information on medicinal herbal pharmacology.

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“…Yang et al investigated the ability of Chinese medicinal plants (CMP) components to target serotonin receptor 5-HT1B, which is widely expressed in several of cognitive and psychiatric disorders, such as depression and AD ( Gadgaard and Jensen, 2020 ). Emodin-8-O-β- d -glucopyranoside (EG) active ingredient displayed to be a selective agonist of 5-HT1B activating 5-HT1B-induced signaling pathway and modulating Aβ-related inflammatory process regulation and neural death resistance.…”

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confidence: 99%

Editorial: Natural products as drivers in drug development for neurodegenerative disorders

et al. 2022

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Functional characterization of 5-HT1A and 5-HT1B serotonin receptor signaling through G-protein-activated inwardly rectifying K+ channels in a fluorescence-based membrane potential assay

Cited by 7 publications

References 72 publications

BK Channels Regulate LPS-induced CCL-2 Release from Human Pulmonary Endothelial Cells

BK Channels Regulate LPS-induced CCL-2 Release from Human Pulmonary Endothelial Cells

A Novel 5-HT1B Receptor Agonist of Herbal Compounds and One of the Therapeutic Uses for Alzheimer’s Disease

Editorial: Natural products as drivers in drug development for neurodegenerative disorders

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