PTEN Subcellular Localization Dictates Function

Mutations in phosphatase and tensin homolog (PTEN) drive unregulated activation of the phosphatidylinositol-3-kinase (PI3K) pathway, resulting in neuronal hypertrophy, and are strongly associated with autism spectrum disorder (ASD). Several PTEN mutations alter subcellular localization, yet how localization governs PTEN function in developing neurons remains unclear. Although PTEN has been reported broadly distributed throughout neurons, here, live imaging of HaloTagged PTEN reveals dynamically regulated localization, suggesting spatial control of its signaling. We then used retroviral-mediated genetic manipulation to delete endogenous Pten in developing hippocampal neurons while simultaneously expressing PTEN fused to defined localization motifs, allowing us to directly test how subcellular targeting regulates neuronal morphology. Loss of Pten produces neurons characterized by enlarged somata, more elaborate dendritic arbors, and increased spine density, length, and head area. Nuclear-excluded PTEN fully rescued these phenotypes, whereas targeting PTEN to filopodia via fusion to the FBAR domain of srGAP3 or to the postsynaptic density via Homer1C corrected or corrected all morphological abnormalities in PTEN-deficient neurons and simplified dendritic arborization compared to wild-type. In contrast, nuclear-localized PTEN produced only partial rescue, normalizing soma size and spine head area but not dendritic complexity or spine density. These findings indicate that PTEN acts locally to restrain growth and structural connectivity, whereas regulation of spine head size can be mediated by PTEN both inside and outside the nucleus, potentially through transcriptional or splicing-dependent mechanisms. Together, our results identify subcellular localization as a critical determinant of PTEN function and reveal spatially distinct mechanisms through which PTEN sculpts neuronal development.