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

Abstract

The Rho family of GTPases have important roles in the morphogenesis of the dendritic spines1,2,3 of neurons in the brain and synaptic plasticity4,5,6,7,8,9 by modulating the organization of the actin cytoskeleton10. Here we used two-photon fluorescence lifetime imaging microscopy11,12,13 to monitor the activity of two Rho GTPases—RhoA and Cdc42—in single dendritic spines undergoing structural plasticity associated with long-term potentiation in CA1 pyramidal neurons in cultured slices of rat hippocampus. When long-term volume increase was induced in a single spine using two-photon glutamate uncaging14,15, RhoA and Cdc42 were rapidly activated in the stimulated spine. These activities decayed over about five minutes, and were then followed by a phase of persistent activation lasting more than half an hour. Although active RhoA and Cdc42 were similarly mobile, their activity patterns were different. RhoA activation diffused out of the stimulated spine and spread over about 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.

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Figure 1: Spatiotemporal dynamics of RhoA activation during long-term structural plasticity induced in single spines
Figure 2: Spatiotemporal dynamics of Cdc42 activation during long-term structural plasticity induced in single spines.
Figure 3: Spatial profile of RhoA and Cdc42 activities.
Figure 4: The effect of Rho GTPase inhibition for structural plasticity of spine head enlargement.

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Acknowledgements

We thank K. Hahn, G. Bokock, A. Aplin, S. Soderling, M. Matsuda and Y. Hayashi for plasmids and reagents, S.-J. Lee for CaMKII activation data, S. Soderling, S. Raghavachari, L. van Aelst, Y. Hayashi, H. Kasai and M. Ehlers for discussion, and H. Hedrick and M. Patterson for comments on the manuscript. We also thank A. Wan for preparing cultured slices and D. Kloetzer for laboratory management. This study was funded by the Howard Hughes Medical Institute, the National Institute of Mental Health, the National Institute of Neurological Disorders and Stroke, the National Institute of Drug Abuse, the Alzheimer’s Association and the Japan Society for the Promotion of Science (H.M.).

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H.M. and R.Y. designed the experiments. H.M. performed the experiments and data analysis. H.W. performed electrophysiological experiments. H.M. and R.Y. wrote the paper.

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Correspondence to Ryohei Yasuda.

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The authors declare no competing financial interests.

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Murakoshi, H., Wang, H. & Yasuda, R. Local, persistent activation of Rho GTPases during plasticity of single dendritic spines. Nature 472, 100–104 (2011). https://doi.org/10.1038/nature09823

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