| Chapter title |
Phenotyping Cellular Viability by Functional Analysis of Ion Channels: GlyR-Targeted Screening in NT2-N Cells
|
|---|---|
| Chapter number | 16 |
| Book title |
Cell Viability Assays
|
| Published in |
Methods in molecular biology, January 2017
|
| DOI | 10.1007/978-1-4939-6960-9_16 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-6959-3, 978-1-4939-6960-9
|
| Authors |
Katharina Kuenzel, Sepideh Abolpour Mofrad, Daniel F. Gilbert |
| Editors |
Daniel F. Gilbert, Oliver Friedrich |
| Abstract |
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. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 6 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 2 | 33% |
| Student > Bachelor | 1 | 17% |
| Student > Doctoral Student | 1 | 17% |
| Student > Master | 1 | 17% |
| Unknown | 1 | 17% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 2 | 33% |
| Chemical Engineering | 1 | 17% |
| Pharmacology, Toxicology and Pharmaceutical Science | 1 | 17% |
| Medicine and Dentistry | 1 | 17% |
| Unknown | 1 | 17% |