Interaction of GAT1 with sodium ions: from efficient recruitment to stabilisation of substrate and conformation DOI Creative Commons
Erika Lazzarin, Ralph Gradisch,

Sophie M.C. Skopec

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: Oct. 13, 2023

Abstract The human GABA transporter (GAT1) is a membrane that mediates the reuptake of neurotransmitter from synaptic cleft into neurons and glial cells. Dysregulation transport cycle has been associated with epilepsy neuropsychiatric disorders, highlighting crucial role in maintaining homeostasis brain levels. GAT1 secondary active couples movement substrate to simultaneous sodium chloride ions along their electrochemical gradients. Using MD simulations, we identified novel recruiting site at entrance outer vestibule, which attracts positively charged increases local concentration, thereby indirectly increasing affinity. Mutations negatively residues slowed binding kinetics, while experimental data revealed change dependency uptake reduction Simulation showed displays higher affinity for NA2, plays stabilisation outward-open conformation. We directly show presence ion bound NA2 stability closed inner gate restrains motions TM5. find only weakly NA1 absence GABA, strengthens interaction due completed coordinating shell, explaining cooperativity between sodium.

Language: Английский

Ways of modulating GABA transporters to treat neurological disease DOI Creative Commons
Jonas S. Mortensen,

Amalie N.L. Mikkelsen,

Petrine Wellendorph

et al.

Expert Opinion on Therapeutic Targets, Journal Year: 2024, Volume and Issue: 28(7), P. 529 - 543

Published: July 2, 2024

Introduction The main inhibitory neurotransmitter in the central nervous system (CNS), γ-aminobutyric acid (GABA), is involved a multitude of neurological and psychiatric disorders characterized by an imbalance excitatory signaling. Regulation extracellular levels GABA maintained four transporters (GATs; GAT1, GAT2, GAT3, BGT1), Na+/Cl−-coupled solute carrier 6 (SLC6) family. Despite mounting evidence for involvement non-GAT1 diseases, only GAT1 has successfully been translated into clinical practice via drug tiagabine.

Language: Английский

Citations

4
Clinical and genetic analysis of epilepsy with myoclonic-atonic seizures caused by SLC6A1 gene variant DOI Creative Commons
Zhen Li,

Changming Han,

Hongwei Zhao

et al.

Frontiers in Pediatrics, Journal Year: 2025, Volume and Issue: 12

Published: Jan. 21, 2025

Objective This research intends to examine the clinical characteristics and genetic diversity of a child experiencing epilepsy with myoclonic-atonic seizures (EMAS) attributed variant in SLC6A1 gene. Methods A male diagnosed EMAS underwent electroencephalographic evaluation. Peripheral blood samples were collected for DNA extraction subsequent whole-exon gene sequencing. For previously identified patients, high-throughput sequencing was utilized, whereas Sanger employed parents determine site mutation connection between genotype phenotype. Results The showed delays intellectual language development before disease began. At 1 year 2 months, he had febrile seizures, which succeeded by at years 9 months; these presented as generalized tonic-clonic, myoclonic, along symptoms showing inattention hyperactivity. After receiving treatment levetiracetam (50 mg·kg·d −1 ), has been free last 8 months. Genetic analysis indicated heterozygous missense c.263T > C (p.L88P) child, recognized spontaneous that not documented literature. Conclusion is implicated one etiological factors contributing coupled neurodevelopmental abnormalities. identification this novel enriches spectrum known variants.

Language: Английский

Citations

0
GABA transporter 1 is a promising drug target for CUL4B mutation-associated epilepsy DOI
Wei Jiang, Yanyan Ma, Yufeng Wang

et al.

Acta Pharmacologica Sinica, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 21, 2025

Language: Английский

Citations

0
Lateral Ventricular Neural Stem Cells Provide Negative Feedback to Circuit Activation Through GABAergic Signaling DOI Creative Commons
Moawiah M. Naffaa, Henry H. Yin

Cells, Journal Year: 2025, Volume and Issue: 14(6), P. 426 - 426

Published: March 13, 2025

Recent studies have demonstrated that circuit activation in vivo can regulate proliferation of lateral ventricular neural stem cells (LV NSCs), although the underlying molecular and cellular mechanisms are not yet fully understood. Here, we investigated role GABAergic signaling interaction between LV NSCs anterior cingulate cortex-subependymal-choline acetyltransferase+ (ChAT+) neuron (ACC-subep-ChAT+) circuit. We found monoamine oxidase B (MAOB), a key enzyme involved gamma-aminobutyric acid (GABA) synthesis, is expressed NSCs, ACC-subep-ChAT+ modulate MAOB activity. Additionally, express LRRC8D, core component volume-regulated anion channels, GABA transporter-1 (GAT-1, SLC6A1). show evidence that, through signaling, LRRC8D GAT-1 provide negative feedback signal to ChAT+ neurons, NSCs. These findings suggest MAOB-driven LRRC8D-regulated chloride transport, GAT-1-facilitated reuptake influence NSC dynamics LV.

Language: Английский

Citations

0
Careful Examination of a Novel Azobenzene Paroxetine Derivative and Its Interactions With Biogenic Amine Transporters DOI Creative Commons
Dominik Dreier, Oliver John V. Belleza, Katharina Schlögl

et al.

Journal of Neurochemistry, Journal Year: 2025, Volume and Issue: 169(4)

Published: April 1, 2025

ABSTRACT The serotonin transporter (SERT) belongs to the family of neurotransmitter sodium symporters (NSS), together with other transporters for norepinephrine, dopamine, glycine, and GABA. main physiological role SERT is retrieval previously released from synaptic cleft. Thereby, plays an important in regulating extracellular concentration maintaining serotonergic neurotransmission. This process can be influenced by molecules acting as uptake inhibitors, like paroxetine. Here, we report development a novel photoswitchable paroxetine derivative its pharmacological interaction profile tool compound light‐induced control SERT. Based on azo‐extension strategy, moiety was formed at former position fluoro substituent resulting azo‐paroxetine ( 9 ) easily reversibly switched between active Z inactive E configurations remained stable these configurations: inhibited more than 12 times potently )‐configuration having sub μM IC 50 value. supported electrophysiological patch‐clamp recordings whole‐cell configuration docking studies. No significant toxic impact no off‐target activity norepinephrine (NET), human GABA subtypes 1 3, rat GAT1 were observed. Our results demonstrate that manipulated optopharmacological agent ). thus applied selective manipulation central or peripheral investigations, further benefiting low probability compound‐related effects. image

Language: Английский

Citations

0
Unveiling the crucial role of betaine: modulation of GABA homeostasis via SLC6A1 transporter (GAT1) DOI Creative Commons
Manan Bhatt, Erika Lazzarin, Ana Sofia Alberto‐Silva

et al.

Cellular and Molecular Life Sciences, Journal Year: 2024, Volume and Issue: 81(1)

Published: June 17, 2024

Betaine is an endogenous osmolyte that exhibits therapeutic potential by mitigating various neurological disorders. However, the underlying cellular and molecular mechanisms responsible for its neuroprotective effects remain puzzling.In this study, we describe a possible mechanism behind positive impact of betaine in preserving neurons from excitotoxicity. Here demonstrate at low concentration modulates GABA uptake GAT1 (slc6a1), predominant transporter central nervous system. This modulation occurs through temporal inhibition transporter, wherein prolonged occupancy impedes swift transition to inward conformation. Importantly, modulatory effect on reversible, as blocking disappears with increased extracellular GABA. Using electrophysiology, mass spectroscopy, radiolabelled assay, dynamics simulation has dual role GAT1: mM acts slow substrate, µM blocker GABA, when it below K

Language: Английский

Citations

3
Disrupting stroke-induced GAT-1-syntaxin1A interaction promotes functional recovery after stroke DOI Creative Commons
Yu‐Hui Lin, Feng Wu, Tingyou Li

et al.

Cell Reports Medicine, Journal Year: 2024, Volume and Issue: 5(11), P. 101789 - 101789

Published: Oct. 17, 2024

Although stroke is a frequent cause of permanent disability, our ability to promote recovery limited. Here, we design small-molecule promoting agent that works by dissociating γ-aminobutyric acid (GABA) transporter 1 (GAT-1) from syntaxin1A (Synt1A), soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) protein. Stroke induces an increase in GAT-1-Synt1A interaction the subacute phase, critical period for functional recovery. Uncoupling reverses stroke-induced GAT-1 dysfunction and cortical excitability decline enhances synaptic GABAergic inhibition consequently oscillations network plasticity facilitating assembly SNARE complex at synapse. Based on molecular mechanism binding Synt1A, blockers. Among them, ZLQ-3 exhibits greatest potency. Intranasal use ZLQ-3-1, glycosylation product ZLQ-3, substantially lessens impairments sensorimotor cognitive functions rodent models. This compound, or its analogs, may serve as

Language: Английский

Citations

3
The epigenetic signatures of opioid addiction and physical dependence are prevented by D-cysteine ethyl ester and betaine DOI Creative Commons
Jennifer McDonough, Naveen Kumar Singhal,

Paulina M. Getsy

et al.

Frontiers in Pharmacology, Journal Year: 2024, Volume and Issue: 15

Published: Aug. 30, 2024

We have reported that D,L-thiol esters, including D-cysteine ethyl ester (D-CYSee), are effective at overcoming opioid-induced respiratory depression (OIRD) in rats. Our on-going studies reveal co-injections of D-CYSee with multi-day morphine injections markedly diminish spontaneous withdrawal usually occurs after cessation multiple Chronically administered opioids known (1) to alter cellular redox status, thus inducing an oxidative state, and (2) for overall decrease DNA methylation, therefore resulting the transcriptional activation previously silenced long interspersed elements (LINE-1) retrotransposon genes. The first objective present study was determine whether one carbon metabolism methyl donor, betaine, would maintain control normal methylation levels human neuroblastoma cell cultures (SH-SY5Y) under overnight challenge (100 nM). second and/or betaine could degree physical dependence male Sprague Dawley data showed treatment reduced GSH levels, induced mitochondrial damage, decreased global increased LINE-1 mRNA expression. These adverse effects by morphine, which diminished reducing capacity compromised maintenance membrane potential SH-SY5Y cells, prevented concurrent application µM) or (300 µM). Furthermore, our demonstrated (250 μmol/kg, IV) a lesser extent, IV), development (escalating daily doses 10-30 mg/kg, as assessed number phenomena elicited injection opioid receptor antagonist, naloxone (1.5 IV). findings provide evidence prevent appearance alterations epigenetic signatures commonly seen neural cells involved dependence/addiction, lessen morphine.

Language: Английский

Citations

2
GABAergic mechanisms in alcohol dependence DOI
Mikko Uusi‐Oukari, Esa R. Korpi

International review of neurobiology, Journal Year: 2024, Volume and Issue: unknown, P. 75 - 123

Published: Jan. 1, 2024

Language: Английский

Citations

2
The kinase LRRK2 is required for the physiological function and expression of the glial glutamate transporter EAAT2 (SLC1A2) DOI Creative Commons
Angela Di Iacovo,

Chiara d’Agostino,

Manan Bhatt

et al.

Journal of Neurochemistry, Journal Year: 2024, Volume and Issue: 169(1)

Published: Dec. 10, 2024

Neurotransmitter transporters (NTTs) control synaptic responses by modulating the concentration of neurotransmitters at cleft. Glutamate is most abundant excitatory neurotransmitter in brain and needs to be finely tuned time space maintain a healthy precise neurotransmission. The glutamate transporter EAAT2 (SLC1A2) primarily responsible for clearance. impairment has been associated with Alzheimer's disease (AD), Huntington's (HD), amyotrophic lateral sclerosis (ALS), Parkinson's (PD). Mutations leucine-rich repeat kinase 2 (LRRK2) contribute both monogenic sporadic forms PD, which common substitution Gly2019Ser significant deficit expression. role pathological mutants LRRK2 intensively studied reviewed. Here we have focused attention on physiological EAAT2, comparing activity NTTs or without kinase. By heterologous expression Xenopus laevis oocytes two-electrode voltage clamp, current amplitudes selected kinetic parameters collected presence absence LRRK2. results show that function are impaired also under its pharmacological inhibition via MLi-2 treatment. stabilizes increasing amount plasma membrane. Interestingly, action EAAT2-specific, as observed no changes transport amplitude obtained other inhibitory studied. This study, first time, demonstrates importance function, highlighting specificity LRRK2-mediated modulation suggesting potential checkpoint preserving neurons from excitotoxicity. In conditions clearance, targeting regulation may offer novel therapeutic opportunities.

Language: Английский

Citations

1