Local, persistent activation of Rho GTPases during plasticity of single dendritic spines

Abstract

The Rho family of GTPases play important roles in morphogenesis of dendritic spines1–3 and synaptic plasticity4–9 by modulating the organization of the actin cytoskeleton10. Here, we monitored the activity of Rho GTPases, RhoA and Cdc42, in single dendritic spines undergoing structural plasticity associated with long-term potentiation (LTP) using 2-photon fluorescence lifetime imaging microscopy (2pFLIM)11–13. When long-term volume increase was induced in a single spine using 2-photon glutamate uncaging14,15, RhoA and Cdc42 were rapidly activated in the stimulated spine. These activities decayed over ~5 min, and were then followed by a phase of persistent activation lasting more than 30 min. Although active RhoA and Cdc42 were similarly mobile, their activity patterns were different. RhoA activation diffused out of the stimulated spine and spread over ~5 μm along the dendrite. In contrast, Cdc42 activation was restricted to the stimulated spine, and exhibited a steep gradient at the spine necks. Inhibition of the Rho-Rock pathway preferentially inhibited the initial spine growth, whereas the inhibition of the Cdc42-Pak pathway blocked the maintenance of sustained structural plasticity. RhoA and Cdc42 activation depended on Ca2+/calmodulin-dependent kinase (CaMKII). Thus, RhoA and Cdc42 relay transient CaMKII activation13 to synapse-specific, long-term signalling required for spine structural plasticity.