Detail of Fig3A_MNI-Glu-Minus

Two-photon RICS images of cultured hippocampal neurons expressing EGFP5 in mouse spine structural without glutamate induction.
Two-photon RICS images of cultured hippocampal neurons expressing EGFP5 in mouse spine structural without glutamate induction.
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193.1 KB

Mus musculus ( NCBITaxon:10090 )
Cell Line
Protein tags

Molecular Function (MF)
signaling molecule ( GO:0048018 )
Biological Process (BP)
actin polymerization ( GO:0030041 ) stimulation of synapse structural plasticity ( GO:0051835 )
Cellular Component (CC)
dendritic spine ( GO:0043197 )
Biological Imaging Method
two-photon laser scanning microscopy ( Fbbi:00000254 )
X scale
0.206 micrometer/pixel
Y scale
0.206 micrometer/pixel
Z scale
T scale
-20, -10, -1, 1, 15, 30 minutes

Image Acquisition
Experiment type
Microscope type
Acquisition mode
Contrast method
Microscope model
Detector model
Objective model
Filter set

Summary of Methods
See details in Obashi K, et. al. (2019) Cell Rep., 27(5):1503-1515.
Related paper(s)

Kazuki Obashi, Atsushi Matsuda, Yasuhiro Inoue, Shigeo Okabe (2019) Precise Temporal Regulation of Molecular Diffusion within Dendritic Spines by Actin Polymers during Structural Plasticity., Cell reports, Volume 27, Number 5, pp. 1503-1515.e8

Published in 2019 Apr 30

(Abstract) The biochemical transduction of excitatory synaptic signals occurs in the cytoplasm within dendritic spines. The associated reaction kinetics are shaped by the mobility of the signaling molecules; however, accurate monitoring of diffusional events within the femtoliter-sized spine structures has not yet been demonstrated. Here, we applied two-photon fluorescence correlation spectroscopy and raster image correlation spectroscopy to monitor protein dynamics within spines, revealing that F-actin restricts the mobility of proteins with a molecular mass of >100 kDa. This restriction is transiently removed during actin remodeling at the initial phase of spine structural plasticity. Photobleaching experiments combined with super-resolution imaging indicate that this increase in mobility facilitates molecular interactions, which may modulate the functions of key postsynaptic signaling molecules, such as Tiam1 and CaMKII. Thus, actin polymers in dendritic spines act as precise temporal regulators of molecular diffusion and modulate signal transduction during synaptic plasticity.
(MeSH Terms)

Shigeo Okabe , the University of Tokyo, Tokyo , Department of Cellular Neurobiology, Graduate School of Medicine
Kazuki Obashi

OMERO Dataset
OMERO Project