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Stability of PSD-95 clusters in the intact mouse neocortex. A sparse subpopulation
of layer 2/3 neurons expressed a red cytoplasmic protein (mCherry1) and PSD-95 fused
to a green fluorescent protein (GFP). Left, overview of a dendritic branch. Right,
time-lapse imaging of a region of interest (white box in the left image). The relatively
stable sizes of the green spots indicate stable relative synapse sizes.
Noah Gray, Robby Weimer, and Karel Svoboda
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Two-photon fluorescence lifetime microscopy of Ras
activation in single dendritic spines in brain slices. A: The imaged neuron
expresses a genetically encoded Ras sensor based on GFP (green fluorescent
protein) and mRFP1 (monomeric red fluorescent protein). The fluorescence
lifetime of the GFP decreases with Ras activation. Color indicates
fluorescence lifetime. At t = 0, glutamate was uncaged next to the spine
marked by the arrowhead. In response, Ras activity increased locally and the
spine grew in volume. B: Ras activity increased over 10 mm of dendritic
length. C: Changes in spine volume in selected regions of interest were
specific to a synapse.
Ryohei Yasuda and Karel Svoboda
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Synaptic input map for a layer 2/3 neuron in the barrel
cortex (colored background). Overlaid is the morpholoy of the neuron (blue)
and electrophysiological traces from which the synaptic input map was
constructed (black). A schematic circuit diagram is also shown (white).
From the Svoboda lab. For details, see Shepherd, G.M., Pologruto, T.A.,
and Svoboda, K. 2003. Neuron 38:277–289.
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Experience-dependent synaptic plasticity in the adult
neocortex in vivo. Left, images collected 24 hours apart. Note stable (yellow
arrow) and transient (blue and red arrows) spines. Scale bar, 5 mm.
Right, electron micrograph of new spine, indicating that it participates in
synaptic transmission. Scale bar, 0.125 mm.
From Trachtenberg, J.T., Chen, B.E., Knott, G.W., Feng, G., Sanes, J.R.,
Welker, E., and Svoboda, K. 2002. Nature 420:788–794. © 2002 Nature
Publishing Group.
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