Myosin-Va Transports the Endoplasmic Reticulum into the Dendritic Spines of Purkinje Neurons

Extension of the endoplasmic reticulum (ER) into dendritic spines of Purkinje neurons (PNs) is required for cerebellar synaptic plasticity and is disrupted in animals with null mutations in Myo5a, the gene encoding myosin-Va(1–3). Notably, the mechanism ensuring the ER's localization to spines has not been unraveled. While it has been proposed that animal class V myosins localize organelles by tethering them to the actin cytoskeleton(4–7), we demonstrate here that myosin-Va acts as a point-to-point organelle transporter to pull ER as cargo into PN spines. Specifically, the myosin accumulates at the ER tip as the organelle moves into spines, and the myosin's ability to hydrolyze ATP is required for spine ER targeting. Moreover, myosin-Va is responsible for the vast majority of spine ER insertional events. Finally, attenuation of the myosin's ability to move along actin filaments reduces the maximum velocity of ER movement into spines, providing direct evidence that myosin-Va drives ER motility. Thus, we establish that an actin-based motor moves ER within animal cells, and we uncover the mechanism that mediates ER localization to PN spines, a prerequisite for synaptic plasticity.