The Journal of Neuroscience, January 4, 2006, ():
Distinct Morphological Stages of Dentate Granule Neuron Maturation in the Adult Mouse Hippocampus
J. Neurosci. Zhao et al.
26: 3
Supplemental data
Files in this Data Supplement:
- supplemental material
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Supplementary Figure 1. Measurement of overall axonal growth. a, Brain sections (one-in-six series) were immunostained with NeuN antibodies to reveal stratum pyramidale and with GFP antibodies to enhance GFP signal. A straight line was drawn to connect the ends of the two blades of the dentate gyrus. The crossing point between the line and pyramidale was used as the starting point, and a tracer was placed following the border between stratum pyramidale and stratum radiatum with ImagePro Measurement function. Arrows indicate where axonal fibers ended at different time points. b, The traced lengths of axonal growth at 7, 10, 11, 12, 14, 16, and 56 dpi. Asterisks indicate significant differences were found between groups by post hoc analysis.
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Supplementary Figure 2. Classification of mushroom spines. a, IGL Trace program (http://synapses.bu.edu/tools/trace/trace.htm) was used to measure the area of the spine head and major/minor axes of the spine head. The estimated surface area was calculated using the two axes. b, The estimated surface area highly correlates with contour measured surface area. c, The distribution of the estimated surface areas of spine heads. A spine is considered as mushroom type when the estimated surface area is ≥ 0.4 μm2. This number was derived from (average+2×Standard deviation) from over 400 spines measured.
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Supplementary Figure 3. Calculation of spine motility. The measure co-localization function of Metamorph was used to calculate spine motility. a, The spine of example was cropped out of the aligned time series. b, The time series of the spine of interest was converted to 4 individual images representing the 4 time points. Each image was binarized with the same threshold level, and differences between time points were obtained through “measure co-localization.” c, Example of data output from “measure co-localization;” the numbers were used in the function to calculate spine motility.
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Supplementary Figure 4. The schematic structure of retroviral vectors and the experimental paradigm. a, Schematic structure of viral vector CAG-GFP. ψ: Viral packaging signal; WPRE: posttranscriptional regulatory element. b, Experimental paradigm. Three groups of animals were used in this study: adult (female C57Bl/6, 7-10 weeks old), P10 (C57Bl/6, postnatal day 10), and adult running (female C57Bl/6, 7-10 weeks old, housed with unlimited access to running wheels starting 7 days before surgery). Arrows indicate the day of stereotaxic surgery. Arrowheads indicate the time points when brains were taken from injected animals after viral infection. Data from the gray arrowheads were only mentioned in the text.
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Supplementary Videos. Representative time series for spine motility studies. a, Adult running, 56 dpi. b, Adult, 126 dpi.
Each movie consists of 4 time frames (0, 30, 60 and 90 min).
- supplemental material
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Supplementary Videos. Representative time series for spine motility studies. a, Adult running, 56 dpi. b, Adult, 126 dpi.
Each movie consists of 4 time frames (0, 30, 60 and 90 min).
