The glycine receptor alpha 2 (GlyRα2) is the only functionally expressed glycine receptor in adult striatal projection neurons (SPNs) and an important modulator of signal integration within the dorsal striatum. SPNs are a convergence point for glutamatergic and dopaminergic input, which is key to reward-motivated behavior. Thus, targeting GlyRα2 offers great potential to modulate striatum-orchestrated behavior. However, the impact of GlyRα2's subcellular localization on SPN activity regulation remains unclear.Neurons are morphologically complex cells, containing many subcellular compartments (somata, branched dendrites, dendritic spines and axon initial segments). Ion channels, such as the GlyRα2, are expressed at different densities in subcellular compartments, whose function varies with localization. Here, we aim to understand the GlyRα2 subcellular localization and dynamics by fusing a genetically-encoded yellow fluorescent protein-based chloride-sensor to a human GlyRα2 (hGlyRα-mClYFP).We confirmed that the hGlyRα-mClYFP retains normal whole-cell current activity whilst possessing mClYFP sensor properties to detect chloride concentration changes. We measured transfected hGlyRα-mClYFP HEK293 cells glycine-induced currents to show that this receptor has normal glycine sensitivity and physiological activity. We measured simultaneously chloride-induced fluorescent quenching in TIRF microscopy to indicate that the fluorescence signal is directly linked to the glycine-induced chloride current. Next, we set up an embryonic cortical-striatal culture and investigated the localization and dynamics of GlyRα2.We present an innovative sensor that allows us to determine dynamic changes in the localization of GlyRα2. This will significantly enhance our understanding of the GlyRα2 in the SPNs and can aid in the development of novel therapies for reward-related pathologies.