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Second Harmonic Generation Imaging of voltage in dendritic spines.

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Dendritic spines mediate most excitatory inputs in the brain. Although it is clear that spines compartmentalize calcium, it is still unknown what role, if any, they play in integrating synaptic inputs. To investigate the electrical function of spines directly we use second harmonic generation (SHG) imaging of membrane potential in pyramidal neurons from hippocampal cultures and neocortical brain slices. With FM 4 -64 as an intracellular SHG chromophore, we image membrane potential in the soma, dendritic branches and spines. The SHG response to voltage is linear and appears based on an electro-optic mechanism. The SHG sensitivity of the chromophore in spines is similar to that of the parent dendritic shaft and the soma. Backpropagation of somatic action potentials generate SHG signals at spines with similar amplitude and kinetics to somatic ones. Our data are the first optical measurements of membrane potential from spines and demonstrate directly that backpropagating action potentials invade the spines.

This talk is part of the Imaging and Mathematics series.

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