The Journal of Neuroscience, August 30, 2006, ():
Cooperative Astrocyte and Dendritic Spine Dynamics at Hippocampal Excitatory Synapses
J. Neurosci. Haber et al.
26: 8881
Supplemental data
Files in this Data Supplement:
- supplemental material
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Supplemental Figure 1. To control for artifacts caused by slight fluctuations in signal intensity or sample photobleaching, multiple dendritic shaft segments (assumed to be stable structures) along each quantified region were used to adjust surface thresholds for spine and astrocytic profile volumes at each time point (See methods). (a) Example of a quantified region of dendrite (red) and the control shaft segments (white, arrowheads). (b) Plot of control shaft segment volume changes over time following threshold adjustments. Scale bar = 1µm.
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Supplemental Figure 2. Localization of GLT-1/EAAT-2 in infected astrocytes expressing EGFPf. (a) Western blot analysis showing the detection of a ~70kD protein from adult hippocampal lysate corresponding to the molecular weight of GLT-1/EAAT-2. (b & c) Control experiments showing the absence of labeling in the absence of primary or secondary antibodies, respectively. (d-g) Localization of GLT-1/EAAT-2 clusters on the processes of a EGFPf-expressing protoplasmic astrocyte. (h & i) Localization of GLT-1/EAAT-2 clusters near dendritic spines. Shown are images of the GLT-1/EAAT-2 labeling before and after performing a three-dimensional “masking” procedure to preserve only the GLT-1/EAAT-2 clusters that are associated with the dendrite (see methods). Scale bars = 10µm in (a-f) and 1µm in (h & i).
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Supplemental Movie 1. Time-lapse movie of Figure 5 showing confocal maximum projections of an EGFPf labeled astrocyte (a), and three-dimensional reconstructions (b) showing process extension and retraction over 10 minutes, plotted in (c). Scale bar = 5 µm.
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Supplemental Movie 2. Time-lapse movie of Figure 6a showing an astrocytic process extending towards a dendritic spine over a 12 minute period. Scale bar = 0.5 µm.
- supplemental material
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Supplemental Movie 3. Time-lapse movie of Figure 6b showing an astrocytic process retracting away from a dendritic spine over a 12 minute period. Scale bar = 0.5 µm.
