title: Optical control of ligand-gated ion channels.
abstract: In the vibrant field of optogenetics, optics and genetic targeting are combined to commandeer cellular functions, such as the neuronal action potential, by optically stimulating light-sensitive ion channels expressed in the cell membrane. One broadly applicable manifestation of this approach are covalently attached photochromic tethered ligands (PTLs) that allow activating ligand-gated ion channels with outstanding spatial and temporal resolution. Here, we describe all steps towards the successful development and application of PTL-gated ion channels in cell lines and primary cells. The basis for these experiments forms a combination of molecular modeling, genetic engineering, cell culture, and electrophysiology. The light-gated glutamate receptor (LiGluR), which consists of the PTL-functionalized GluK2 receptor, serves as a model.
authors.affiliation: Department of Neurology, Yale University School of Medicine, New Haven, CT, USA.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Szobota
authors.forename: Catherine
authors.initials: C
authors.lastname: McKenzie
authors.forename: Harald
authors.initials: H
authors.lastname: Janovjak
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D004210:Dissection; D055724:Electrophysiological Phenomena; D005971:Glutamates / Q000378:metabolism; D057809:HEK293 Cells; D006624:Hippocampus / Q000166:cytology / Q000502:physiology; D006801:Humans; D016296:Mutagenesis; D009154:Mutation; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D062308:Optogenetics / Q000379:methods*; D018408:Patch-Clamp Techniques; D016133:Polymerase Chain Reaction; D051381:Rats; D018092:Kainic Acid Receptors / Q000235:genetics* / Q000378:metabolism; D014162:Transfection; D000097806:GluK2 Kainate Receptor
chemical_list: D005971:Glutamates; D018092:Kainic Acid Receptors; D000097806:GluK2 Kainate Receptor