Summary: Dendritic spine enlargement by synaptic activation is thought to increase synaptic efficacy underlying laguna 3hp dust collector learning and memory.This process requires forces generated by actin polymerization and actin-adhesion coupling (clutch coupling).Here, we describe a protocol to monitor actin filament retrograde flow and actin polymerization within spines using a standard epi-fluorescence microscope.
In combination with chemical long-term potentiation, this protocol allows us to quantify clutch coupling efficiency and actin polymerization rate, which are essential variables for generating forces for activity-dependent rosy teacup dogwood spine enlargement.For complete details on the use and execution of this protocol, please refer to Kastian et al.(2021).