Role of a Galphai2 protein splice variant in the formation of an intracellular dopamine D2 receptor pool

Treatment of D2-receptor-expressing cells with specific drugs upregulates the receptor number at the cell surface independently of protein synthesis, leading to the concept of an intracellular receptor pool. However, how this pool is operating is still an enigma. Here, we report that a splice variant of the Galphai2 protein, protein sGalphai2, plays a crucial role in the maintenance of this D2-receptor pool. Co-expression of sGi2 with D2 receptor reduced receptor localization to cell surface by one-third. This effect is associated with specific intracellular protein-protein interaction and the formation of a sGi2-D2-receptor complex. It has been suggested that the formation of this complex serves to prevent D2 receptors from reaching the cell membrane. Treatment of D2-receptor-expressing cells with agonists increased the number of cell surface D2 receptors and coincided with a reduction in these receptors from intracellular complexes, suggesting that agonist treatment released D2 receptors from the complex allowing them to localize to the cell membrane. Thus, in addition to elucidating how the intracellular pool of D2 receptor functions, our findings uncover a novel mechanism regulating the density of cell surface D2 receptors.