Journal
JOURNAL OF NEUROSCIENCE
Volume 25, Issue 12, Pages 3107-3112Publisher
SOC NEUROSCIENCE
DOI: 10.1523/JNEUROSCI.0085-05.2005
Keywords
Ca2+/calmodulin-dependent protein kinase II; fluorescent resonance energy transfer; two photon laser-scanning microscopy; synaptic plasticity; hippocampus; excitatory amino acid
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Ca2+/calmodulin-dependent protein kinase II ( CaMKII) is highly enriched in excitatory synapses in the CNS and critically involved in synaptic plasticity, learning, and memory. However, the precise temporal and spatial regulation of CaMKII activity in living cells has not been well described, because of a lack of specific methods. We tried to address this by optically detecting the conformational change in CaMKII during activation using fluorescence resonance energy transfer (FRET). The engineered FRET probe Camui alpha detects calmodulin binding and autophosphorylation at threonine 286 that renders the enzyme constitutively active. In combination with two-photon microscopy, we demonstrate that Camui alpha can be used to observe temporal and spatial regulation of CaMKII activity in living neurons.
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