A versatile optical tool for studying synaptic GABA_A receptor trafficking.

ABSTRACT: Live-cell imaging methods can provide critical real-time receptor trafficking measurements. Here, we describe an optical tool to study synaptic γ-aminobutyric acid (GABA) type A receptor (GABA_AR) dynamics through adaptable fluorescent-tracking capabilities. A fluorogen-activating peptide (FAP) was genetically inserted into a GABA_AR γ2 subunit tagged with pH-sensitive green fluorescent protein (γ2^pHFAP). The FAP selectively binds and activates Malachite Green (MG) dyes that are otherwise non-fluorescent in solution. γ2^pHFAP GABA_ARs are expressed at the cell surface in transfected cortical neurons, form synaptic clusters and do not perturb neuronal development. Electrophysiological studies show γ2^pHFAP GABA_ARs respo

SUBMITTER: Lorenz-Guertin JM 

PROVIDER: S-EPMC5702044 | biostudies-literature | 2017 Nov

REPOSITORIES: biostudies-literature