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GFP-calcium-calmodulin-dependent protein kinase (CaMKII) expression and translocation after stimulation with 100 μM glutamate with 10 μM glycine via pipette. (A) GFP-CaMKII expression prior to stimulation. (B) DIC image of the same neuron with measured dendrites highlighted in red (right) and blue (left) and an arrow indicating the position of the micropipette. Scale bar = 50 μm. (C) Time-lapse subtraction images (current fluorescence level minus initial fluorescence level; increases in fluorescence appear whiter while decreases are darker/black). Hollow white arrows indicate dendrite locations that show increased relative fluorescence over time, solid white arrow indicates the location where relative fluorescence level decreases. (D) <t>Linescan</t> measures of the same neuron shown in (C) indicating fluorescence levels for respective dendrites, red (top) and blue (bottom), starting proximal to the cell body. Brackets denote regions that correspond to boxes shown in (A,B) . Black linescan trace indicates fluorescence levels measured at first post-stimulation image capture for comparison. Arrowhead indicates the location of the stimulation pipette.
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GFP-calcium-calmodulin-dependent protein kinase (CaMKII) expression and translocation after stimulation with 100 μM glutamate with 10 μM glycine via pipette. (A) GFP-CaMKII expression prior to stimulation. (B) DIC image of the same neuron with measured dendrites highlighted in red (right) and blue (left) and an arrow indicating the position of the micropipette. Scale bar = 50 μm. (C) Time-lapse subtraction images (current fluorescence level minus initial fluorescence level; increases in fluorescence appear whiter while decreases are darker/black). Hollow white arrows indicate dendrite locations that show increased relative fluorescence over time, solid white arrow indicates the location where relative fluorescence level decreases. (D) <t>Linescan</t> measures of the same neuron shown in (C) indicating fluorescence levels for respective dendrites, red (top) and blue (bottom), starting proximal to the cell body. Brackets denote regions that correspond to boxes shown in (A,B) . Black linescan trace indicates fluorescence levels measured at first post-stimulation image capture for comparison. Arrowhead indicates the location of the stimulation pipette.
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GFP-calcium-calmodulin-dependent protein kinase (CaMKII) expression and translocation after stimulation with 100 μM glutamate with 10 μM glycine via pipette. (A) GFP-CaMKII expression prior to stimulation. (B) DIC image of the same neuron with measured dendrites highlighted in red (right) and blue (left) and an arrow indicating the position of the micropipette. Scale bar = 50 μm. (C) Time-lapse subtraction images (current fluorescence level minus initial fluorescence level; increases in fluorescence appear whiter while decreases are darker/black). Hollow white arrows indicate dendrite locations that show increased relative fluorescence over time, solid white arrow indicates the location where relative fluorescence level decreases. (D) <t>Linescan</t> measures of the same neuron shown in (C) indicating fluorescence levels for respective dendrites, red (top) and blue (bottom), starting proximal to the cell body. Brackets denote regions that correspond to boxes shown in (A,B) . Black linescan trace indicates fluorescence levels measured at first post-stimulation image capture for comparison. Arrowhead indicates the location of the stimulation pipette.
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GFP-calcium-calmodulin-dependent protein kinase (CaMKII) expression and translocation after stimulation with 100 μM glutamate with 10 μM glycine via pipette. (A) GFP-CaMKII expression prior to stimulation. (B) DIC image of the same neuron with measured dendrites highlighted in red (right) and blue (left) and an arrow indicating the position of the micropipette. Scale bar = 50 μm. (C) Time-lapse subtraction images (current fluorescence level minus initial fluorescence level; increases in fluorescence appear whiter while decreases are darker/black). Hollow white arrows indicate dendrite locations that show increased relative fluorescence over time, solid white arrow indicates the location where relative fluorescence level decreases. (D) Linescan measures of the same neuron shown in (C) indicating fluorescence levels for respective dendrites, red (top) and blue (bottom), starting proximal to the cell body. Brackets denote regions that correspond to boxes shown in (A,B) . Black linescan trace indicates fluorescence levels measured at first post-stimulation image capture for comparison. Arrowhead indicates the location of the stimulation pipette.

Journal: Frontiers in Synaptic Neuroscience

Article Title: Locally-Induced CaMKII Translocation Requires Nucleotide Binding

doi: 10.3389/fnsyn.2020.00004

Figure Lengend Snippet: GFP-calcium-calmodulin-dependent protein kinase (CaMKII) expression and translocation after stimulation with 100 μM glutamate with 10 μM glycine via pipette. (A) GFP-CaMKII expression prior to stimulation. (B) DIC image of the same neuron with measured dendrites highlighted in red (right) and blue (left) and an arrow indicating the position of the micropipette. Scale bar = 50 μm. (C) Time-lapse subtraction images (current fluorescence level minus initial fluorescence level; increases in fluorescence appear whiter while decreases are darker/black). Hollow white arrows indicate dendrite locations that show increased relative fluorescence over time, solid white arrow indicates the location where relative fluorescence level decreases. (D) Linescan measures of the same neuron shown in (C) indicating fluorescence levels for respective dendrites, red (top) and blue (bottom), starting proximal to the cell body. Brackets denote regions that correspond to boxes shown in (A,B) . Black linescan trace indicates fluorescence levels measured at first post-stimulation image capture for comparison. Arrowhead indicates the location of the stimulation pipette.

Article Snippet: The density of GFP-CaMKII puncta was calculated by dividing the number of identified puncta by the length of the dendrite measured using the MetaMorph linescan function yielding a value in puncta/μm.

Techniques: Expressing, Translocation Assay, Transferring, Fluorescence