Künzel K, Abolpour Mofrad S, Gilbert D (2017)
Publication Language: English
Publication Type: Book chapter / Article in edited volumes
Publication year: 2017
Publisher: Humana Press
Edited Volumes: Methods in Molecular Biology
Series: Cell Viability Assays, Methods and Protocols
Book Volume: 1601
ISBN: 978-1-4939-6959-3
DOI: 10.1007/978-1-4939-6960-9_16
Glycine receptor chloride channels (GlyRs) are attractive drug targets for therapeutic intervention and are also more and more recognized in the context of in vitro neurotoxicity and developmental neurotoxicity testing. Assaying the functional properties of GlyR can serve as an indicator of cellular viability and the integrity of the developing and mature central nervous system. Human pluripotent NTERA-2 (NT2) stem cells undergo neuronal differentiation upon stimulation with retinoic acid and express a large variety of neuronal proteins-including GlyR. YFP-I152L, a halide-sensitive variant of yellow fluorescent protein, allows high-throughput fluorescence-based functional analysis of GlyRs in NT2 cells. Here we describe a protocol for phenotyping of cellular viability by functional analysis of GlyR in neuronally differentiated NT2 (NT2-N) cells using YFP-I152L as a reporter of functional integrity of GlyRs. The protocol describes neuronal differentiation of NT2 stem cells, transient transfection of NT2-N cells with YFP-I152L as well as functional imaging and analysis of data from high-content imaging.
APA:
Künzel, K., Abolpour Mofrad, S., & Gilbert, D. (2017). Phenotyping Cellular Viability by Functional Analysis of Ion Channels: GlyR-Targeted Screening in NT2-N Cells. In Humana Press (Eds.), Methods in Molecular Biology. Humana Press.
MLA:
Künzel, Katharina, Sepideh Abolpour Mofrad, and Daniel Gilbert. "Phenotyping Cellular Viability by Functional Analysis of Ion Channels: GlyR-Targeted Screening in NT2-N Cells." Methods in Molecular Biology. Ed. Humana Press, Humana Press, 2017.
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