Induction of input-specific spine shrinkage on dendrites of rodent hippocampal CA1 neurons using two-photon glutamate uncaging

Shrinkage and loss of dendritic spines are vital components of the neuronal plasticity that supports learning. To investigate the mechanisms of spine shrinkage and loss, Oh and colleagues established a two-photon glutamate uncaging protocol that reliably induces input-specific spine shrinkage on dendrites of rodent hippocampal CA1 pyramidal neurons. Here, we provide a detailed description of that protocol and also an optimized version that can be used to induce input- and synapse-specific shrinkage of dendritic spines at physiological Ca2+ levels. For complete details on the use and execution of this protocol, please refer to Oh et al. (2013), Stein et al. (2015), Stein et al. (2020), and Stein et al. (2021). [Display omitted] •Two-photon photolysis of caged glutamate to induce dendritic spine shrinkage•Live imaging of sparsely labeled neurons using time-lapse two-photon microscopy•Quantitative analysis of dendritic spine size using fluorescence measurements•Preparation and transfection of organotypic hippocampal slice cultures Shrinkage and loss of dendritic spines are vital components of the neuronal plasticity that supports learning. To investigate the mechanisms of spine shrinkage and loss, Oh and colleagues established a two-photon glutamate uncaging protocol that reliably induces input-specific spine shrinkage on dendrites of rodent hippocampal CA1 pyramidal neurons. Here, we provide a detailed description of that protocol and also an optimized version that can be used to induce input- and synapse-specific shrinkage of dendritic spines at physiological Ca2+ levels.