A Recombinant Human Pluripotent Stem Cell Line Stably Expressing Halide-Sensitive YFP-I152L for GABAAR and GlyR-Targeted High-Throughput Drug Screening and Toxicity Testing.

Künzel K, Friedrich O, Gilbert D (2016)

Publication Language: English

Publication Status: Published

Publication Type: Journal article, Original article

Publication year: 2016

Journal

Frontiers in Molecular Neuroscience Frontiers Research Foundation / Frontiers Media

Publisher: Frontiers Research Foundation / Frontiers Media

Book Volume: 9

Article Number: 51

DOI: 10.3389/fnmol.2016.00051

Abstract

GABARs and GlyRs are considered attractive drug targets for therapeutic intervention and are also increasingly recognized in the context of in vitro neurotoxicity (NT) and developmental neurotoxicity (DNT) testing. However, systematic human-specific GABAR and GlyR-targeted drug screening and toxicity testing is hampered due to lack of appropriate in vitro models that express native GABARs and GlyRs. We have established a human pluripotent stem cell line (NT2) stably expressing YFP-I152L, a halide-sensitive variant of yellow fluorescent protein (YFP), allowing for fluorescence-based functional analysis of chloride channels. Upon stimulation with retinoic acid, NT2 cells undergo neuronal differentiation and allow pharmacological and toxicological evaluation of native GABARs and GlyRs at different stages of brain maturation. We applied the cell line in concentration-response experiments with the neurotransmitters GABA and glycine as well as with the drugs strychnine, picrotoxin, fipronil, lindane, bicuculline, and zinc and demonstrate that the established in vitro model is applicable to GABAR and GlyR-targeted pharmacological and toxicological profiling. We quantified the proportion of GABAR and GlyR-sensitive cells, respectively, and identified percentages of approximately 20% each within the overall populations, rendering the cells a suitable model for systematic in vitro GABAR and GlyR-targeted screening in the context of drug development and NT/DNT testing.

Authors with CRIS profile

Katharina Künzel Lehrstuhl für Medizinische Biotechnologie Oliver Friedrich Lehrstuhl für Medizinische Biotechnologie Daniel Gilbert Lehrstuhl für Medizinische Biotechnologie

Related research project(s)

Developing an experimental/analytical optical process for studying expression control of glycine receptor chloride channels - identification of lead molecules and biomarkers for diagnostics and therapy of neurological disorders Oct. 1, 2016 - Sept. 30, 2017 Design of a recombinant human stem cell line for the development of a novel, animal-free DNT toxicity test April 1, 2013 - March 31, 2015

How to cite

APA:

Künzel, K., Friedrich, O., & Gilbert, D. (2016). A Recombinant Human Pluripotent Stem Cell Line Stably Expressing Halide-Sensitive YFP-I152L for GABAAR and GlyR-Targeted High-Throughput Drug Screening and Toxicity Testing. Frontiers in Molecular Neuroscience, 9. https://doi.org/10.3389/fnmol.2016.00051

MLA:

Künzel, Katharina, Oliver Friedrich, and Daniel Gilbert. "A Recombinant Human Pluripotent Stem Cell Line Stably Expressing Halide-Sensitive YFP-I152L for GABAAR and GlyR-Targeted High-Throughput Drug Screening and Toxicity Testing." Frontiers in Molecular Neuroscience 9 (2016).

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