Abstract
We have constructed stable HEK293 cell lines expressing the rat ionotropic glutamate receptor subtypes GluR1i, GluR2Qi, GluR3i, GluR4i, GluR5Q and GluR6Q and characterised the pharmacological profiles of the six homomeric receptors in a fluorescence-based high throughput screening assay using Fluo-4/AM as a fluorescent Ca2+ indicator. In this assay, the pharmacological properties of nine standard GluR ligands correlated nicely with those previously observed in electrophysiology studies of GluRs expressed in Xenopus oocytes or mammalian cells. The potencies and efficacies displayed by the agonists (S)-glutamate, (S)-quisqualate, kainate, (RS)-AMPA, (RS)-ATPA, (RS)-ACPA] and (S)-4-AHCP at the six GluRs were in concordance with electrophysiological studies. Furthermore, the Ki values exhibited by the competitive antagonists NBQX and (RS)-ATPO were also in agreement with findings of previous studies. Finally, the effects of various concentrations of Ca2+ in the assay buffer and of the allosteric modulators cyclothiazide and concanavalin A on GluR signalling were examined. This study represents the most elaborate functional characterisation of multiple AMPA and KA receptor subtypes in the same assay reported to date. We propose that high throughput screening of compound libraries at the six GluRHEK293 cell lines could be helpful in the search for structurally and pharmacologically novel ligands acting at the receptors.
Keywords: High throughput screening, glutamate, AMPA, kainate, ion channel, HTS, Fluo-4, ionotropic
Combinatorial Chemistry & High Throughput Screening
Title: Functional Characterisation of Homomeric Ionotropic Glutamate Receptors GluR1-GluR6 in a Fluorescence-Based High Throughput Screening Assay
Volume: 9 Issue: 2
Author(s): Mette Strange, Hans Brauner-Osborne and Anders A. Jensen
Affiliation:
Keywords: High throughput screening, glutamate, AMPA, kainate, ion channel, HTS, Fluo-4, ionotropic
Abstract: We have constructed stable HEK293 cell lines expressing the rat ionotropic glutamate receptor subtypes GluR1i, GluR2Qi, GluR3i, GluR4i, GluR5Q and GluR6Q and characterised the pharmacological profiles of the six homomeric receptors in a fluorescence-based high throughput screening assay using Fluo-4/AM as a fluorescent Ca2+ indicator. In this assay, the pharmacological properties of nine standard GluR ligands correlated nicely with those previously observed in electrophysiology studies of GluRs expressed in Xenopus oocytes or mammalian cells. The potencies and efficacies displayed by the agonists (S)-glutamate, (S)-quisqualate, kainate, (RS)-AMPA, (RS)-ATPA, (RS)-ACPA] and (S)-4-AHCP at the six GluRs were in concordance with electrophysiological studies. Furthermore, the Ki values exhibited by the competitive antagonists NBQX and (RS)-ATPO were also in agreement with findings of previous studies. Finally, the effects of various concentrations of Ca2+ in the assay buffer and of the allosteric modulators cyclothiazide and concanavalin A on GluR signalling were examined. This study represents the most elaborate functional characterisation of multiple AMPA and KA receptor subtypes in the same assay reported to date. We propose that high throughput screening of compound libraries at the six GluRHEK293 cell lines could be helpful in the search for structurally and pharmacologically novel ligands acting at the receptors.
Export Options
About this article
Cite this article as:
Strange Mette, Brauner-Osborne Hans and Jensen A. Anders, Functional Characterisation of Homomeric Ionotropic Glutamate Receptors GluR1-GluR6 in a Fluorescence-Based High Throughput Screening Assay, Combinatorial Chemistry & High Throughput Screening 2006; 9 (2) . https://dx.doi.org/10.2174/138620706775541918
DOI https://dx.doi.org/10.2174/138620706775541918 |
Print ISSN 1386-2073 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5402 |
Call for Papers in Thematic Issues
Artificial Intelligence Methods for Biomedical, Biochemical and Bioinformatics Problems
Recently, a large number of technologies based on artificial intelligence have been developed and applied to solve a diverse range of problems in the areas of biomedical, biochemical and bioinformatics problems. By utilizing powerful computing resources and massive amounts of data, methods based on artificial intelligence can significantly improve the ...read more
Eco-friendly Agents for Biological Control of Pathogenic Diseases
The discovery of an alternative biological approach to disease management includes work on medicinal products derived from natural sources as a starting point for the development of eco-friendly agents for these diseases and the injuries they cause, as well as reducing human contact with hazardous chemicals and their residues. We ...read more
Emerging trends in diseases mechanisms, noble drug targets and therapeutic strategies: focus on immunological and inflammatory disorders
Recently infectious and inflammatory diseases have been a key concern worldwide due to tremendous morbidity and mortality world Wide. Recent, nCOVID-9 pandemic is a good example for the emerging infectious disease outbreak. The world is facing many emerging and re-emerging diseases out breaks at present however, there is huge lack ...read more
Exploring Spectral Graph Theory in Combinatorial Chemistry
Scope of the Thematic Issue: Combinatorial chemistry involves the synthesis and analysis of a large number of diverse compounds simultaneously. Traditional methods rely on brute force experimentation, which can be time-consuming and resource-intensive. Spectral Graph Theory, a branch of mathematics dealing with the properties of graphs in relation to the ...read more
- Author Guidelines
- Graphical Abstracts
- Fabricating and Stating False Information
- Research Misconduct
- Post Publication Discussions and Corrections
- Publishing Ethics and Rectitude
- Increase Visibility of Your Article
- Archiving Policies
- Peer Review Workflow
- Order Your Article Before Print
- Promote Your Article
- Manuscript Transfer Facility
- Editorial Policies
- Allegations from Whistleblowers
Related Articles
-
Metabolic Targeting of Cancers: From Molecular Mechanisms to Therapeutic Strategies
Current Medicinal Chemistry An Emerging Antiarrhythmic Target: Late Sodium Current
Current Pharmaceutical Design Quantitative Molecular Imaging of Neuronal Nicotinic Acetylcholine Receptors in the Human Brain with A-85380 Radiotracers
Current Medical Imaging Cyclosporin and Organ Specific Toxicity: Clinical Aspects, Pharmacogenetics and Perspectives
Current Clinical Pharmacology Nanoemulgel: A Promising Phase in Drug Delivery
Current Pharmaceutical Design Interaction of Different Proteins with GABA<sub>A</sub> Receptor and their Modulatory Effect on Inhibitory Neural Transmission Leads to Epilepsy
CNS & Neurological Disorders - Drug Targets Atypical GTPases as Drug Targets
Anti-Cancer Agents in Medicinal Chemistry Dietary Approaches and Supplements in the Prevention of Cognitive Decline and Alzheimer's Disease
Current Pharmaceutical Design Conotoxins: Structure, Therapeutic Potential and Pharmacological Applications
Current Pharmaceutical Design Nonsynaptic Receptors for GABA and Glutamate
Current Topics in Medicinal Chemistry Propofol: Therapeutic Indications and Side-Effects
Current Pharmaceutical Design Pathophysiology of Transmissible Spongiform Encephalopathies
Current Medicinal Chemistry - Immunology, Endocrine & Metabolic Agents Current State of Development of Genome Analysis in Livestock
Current Genomics Non-Selective Cation Channel Blockers: Potential Use in Nervous System Basic Research and Therapeutics
Mini-Reviews in Medicinal Chemistry Establishing Genomic/Transcriptomic Links Between Alzheimer’s Disease and Type 2 Diabetes Mellitus by Meta-Analysis Approach
CNS & Neurological Disorders - Drug Targets Age-Specific Impairment of Morris Water Maze Performance Following Neonatal Exposure to Magnetic Resonance Image in Rats
Current Neurovascular Research Commentary [Research Highlights (Making Sense Out of Antisense in Huntington’s Disease)]
CNS & Neurological Disorders - Drug Targets Microemulsions and Nanoemulsions for Targeted Drug Delivery to the Brain
Current Nanoscience Methylphenidate (Ritalin): What Makes it so Widely Prescribed During the Last 60 Years?
Current Drug Therapy Sympathetic Nervous System Dysfunction in Multiple Sclerosis, Linking Neurodegeneration to a Reduced Response to Therapy
Current Pharmaceutical Design