Related Experiment Video
Updated: Jun 12, 2025
Imaging pHluorin-tagged Receptor Insertion to the Plasma Membrane in Primary Cultured Mouse Neurons
Published on: November 20, 2012
Protocol for live neuron imaging analysis of basal surface fraction and dynamic availability of the dopamine
Elnaz Khezerlou1, Jacqueline Saenz2, Sanjana Surya Prakash3
1Department of Neuroscience and Cell Biology, Rutgers University Robert Wood Johnson Medical School, 675 Hoes Lane West, Piscataway, NJ 08854, USA; Rutgers Addiction Research Center, Rutgers Brain Health Institute, 683 Hoes Lane West, Piscataway, NJ 08854, USA.
Abstract:
Surface availability of the dopamine (DA) transporter (DAT) critically influences DA transmission. Here, we present a protocol that describes the preparation of mouse ventral midbrain neurons, the expression of a new optical sensor, DAT-pHluorin, and the utilization of this sensor to analyze the surface availability of DAT in live neurons via fluorescent microscopy. This approach allows quantitative measures of basal surface DAT fraction under genetic backgrounds of interest and live trafficking of DAT in response to psychoactive substances. For complete details on the use and execution of this protocol, please refer to Saenz et al.1.
