The Journal of Neuroscience, May 31, 2006, ():
The Stop Signal Revised: Immature Cerebellar Granule Neurons in the External Germinal Layer Arrest Pontine Mossy Fiber Growth
J. Neurosci. Manzini et al.
26: 6040
Supplemental data
Files in this Data Supplement:
- supplemental material
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Supplementary Figure 1. Granule cells form synapses with each other in culture, mostly after 4 div. Cultures of purified granule cells were immunostained for the synaptic vesicle marker synapsin I (in red) and the post-synaptic density marker PSD-95 (in green) to analyze formation of synaptic contacts during differentiation. At 1 div very little synapsin I and PSD-95 can be observed in immature, EGL-like cultures and there is almost no colocalization of synapsin I and PSD-95. Note how flat cells (marked by asterisks) do not express synapsin I or PSD-95. At 4 div, after granule cells have differentiated into an IGL-like population synapse formation is taking place, shown by colocalization of synapsin I and PDS-95 (yellow puncta in the lower “merge” panel). Scale bar: 20µm.
- supplemental material
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Supplementary Figure 2. Mossy fibers do not extend on flat EGL-like granule cells. GFP-positive mossy fibers from pontine explants prepared from GFP transgenic mice, do not grow on flat granule cells (asterisks), but extend on more differentiated bipolar cells (triangles) in the same culture. Granule cell morphology is shown by DIC on the left-hand panel and the pattern of outgrowth of GFP-positive axons is evident in the center panel. Scale bar: 20 µm. The lower right-hand corner of the GFP panel is magnified on the right to demonstrate how mossy fiber growth cones apposed to immature granule cells do not show signs of retraction or collapse.
