Model
Digital Document
Publisher
Florida Atlantic University
Description
Dendritic spines are the major sites for receiving excitatory synaptic inputs and play important roles in neuronal signal transduction, memory storage and neuronal circuit organization. Structural plasticity of dendritic spines is correlated with functional plasticity, and is critical for learning and memory. Visualization of the changes of dendritic spines at the ultrastructural level that specifically correlated with their function changes in high throughput would shed light on detailed mechanisms of synaptic plasticity.
Here we developed a correlative light and electron microscopy workflow which combines two-photon MNI-glutamate uncaging, pre-embedding immunolabeling, Automatic Tape-collecting Ultramicrotome sectioning and scanning electron microscopy imaging. This method bridges two different visualization platforms, directly linking ultrastructure and function at the level of individual synapses. With this method, we successfully relocated single dendritic spines that underwent long-term potentiation (LTP) induced by two-photon MNI-glutamate uncaging, and visualized their ultrastructures and AMPA receptors distribution at different phases of LTP in high throughput.
Here we developed a correlative light and electron microscopy workflow which combines two-photon MNI-glutamate uncaging, pre-embedding immunolabeling, Automatic Tape-collecting Ultramicrotome sectioning and scanning electron microscopy imaging. This method bridges two different visualization platforms, directly linking ultrastructure and function at the level of individual synapses. With this method, we successfully relocated single dendritic spines that underwent long-term potentiation (LTP) induced by two-photon MNI-glutamate uncaging, and visualized their ultrastructures and AMPA receptors distribution at different phases of LTP in high throughput.
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