Signalling through AMPA receptors on oligodendrocyte precursors promotes myelination by enhancing oligodendrocyte survival
- University College London, United Kingdom
- Iwate Medical University, Japan
- University of Heidelberg, Germany
- Deutches Krebforschungzentrum, University of Heidelberg, Germany
Figures
Figure 1
GluA2–4, but not GluA1 are expressed by presumptive OL lineage cells in the corpus callosum.
(A) Low-magnification images of Gria1 and Gria2 ISH at P14. The corpus callosum (rectangles) is shown at higher magnification for Gria1-4 (left to right) at P1 (B) and P14 (C). Gria2, Gria3 and Gria4…
Figure 2
Gria2-4 are expressed in OL lineage cells.
(A–C) Fluorescence ISH for Gria2-4 followed by immunohistochemistry for Olig2 in P14 corpus callosum indicates that Gria2-4 are expressed in OL lineage cells (arrows). Occasional Gria+ Olig2– cells …
Figure 3
Gria3 germline KO mice generate normal numbers of OPs and OLs.
(A, B) Immunolabelling for Pdgfra+ OPs (A) or CC1+ OLs (B) in P14 corpus callosum of wild type (WT) and Gria3null mice. (C) EdU was administered to P14 wild type and Gria3null mice for 8 hr before …
Figure 4 with 2 supplements
Gria2 knock-out renders AMPARs in OPs inwardly-rectifying but does not alter numbers of OL lineage cells in white matter.
(A) Patch-clamp recording of an OP in SCWM in an acute slice of wild type mouse forebrain, showing inward current generated by bath application of 100 μM kainate. (B) Currents evoked by voltage …
Figure 4—figure supplement 1
Breeding strategies.
(A) Obtaining Gria2 cKO in OL lineage cells. (B) Obtaining Gria3null2–/– double knockouts in OL lineage cells. (C) Obtaining Gria3null2–/–4–/– triple knockouts in OL lineage cells. (?) indicates …
Figure 4—figure supplement 2
Sox10-Cre drives recombination in OL lineage cells in the sub-cortical white matter and cerebral cortex.
Coronal sections of P14 Sox10-Cre: Rosa-YFP mouse forebrain were double-immunolabelled with anti-Sox10 and anti-GFP/YFP. (A) In the corpus callosum (within the dashed lines) the great majority of …
Figure 5 with 2 supplements
The kainate- evoked current in OPs is reduced in Gria3null2–/– mice.
(A) The kainate-evoked current is blocked reversibly by the AMPAR antagonist GYKI-52466 (GYKI, 50 μM). (B) Mean kainate-evoked current density is ∼47% less in Gria3null2–/– OPs compared to Gria3null2…
Figure 5—figure supplement 1
Gria2 is successfully knocked out in Gria3null2–/– mice.
(A) Current-voltage plot showing inward rectification in OPs of P14 Gria3null2–/– mice but not Gria3null2+/– or Gria3null2+/+ when 100 μM spermine is included in the patch pipette. (B) Rectification …
Figure 5—figure supplement 2
Gria1 is not up-regulated in cells in the corpus callosum of Gria3null2–/– mice.
ISH for Gria1 in the P14 corpus callosum of Gria3null2–/– and Gria3null2+/+ mice provides no evidence for compensatory up-regulation of Gria1. Scale bar: 100 μm.
Figure 6
AMPAR-mediated synaptic input is reduced in Gria3null2–/– mice.
(A) EPSCs evoked by minimal stimulation in OPs in the SCWM of P14 Gria3null2+/– and Gria3null2–/– mice. Black triangle marks the time point of stimulation. (B) The amplitude of minimal …
Figure 7 with 2 supplements
Gria3null2–/– mice generate fewer OLs in the corpus callosum.
All data at P14 unless otherwise stated. (A) Immunolabelling for Olig2 and CC1 antigen visualizes differentiated OLs in Gria3null2+/+ and Gria3null2–/– mice. (B) The density of CC1+ OLs is …
Figure 7—figure supplement 1
The division rate of OPs in the corpus callosum is not affected in Gria3null2–/– mice.
(A) Pdgfra immunolabelling and EdU detection in the SCWM of P14 Gria3null2+/+ and Gria3null2–/– mice that were labelled with EdU for 8 hr before fixation. (B) There were no significant changes in …
Figure 7—figure supplement 2
mRNA encoding GluA2-4 continue to be expressed in Enpp6+ early-differentiating OLs.
Double fluorescence ISH for Gria2-4 and Enpp6 followed by immunolabelling for Olig2 in wild type P12 corpus callosum. Many Enpp6+ cells co-express Gria2-4 (arrowheads). Scale bar: 50 μm.
Figure 8
Less myelin in Gria3null2–/– corpus callosum.
(A) Transmission electron micrograph of P14 corpus callosum from Gria3null2+/+ and Gria3null2–/– mice. Scale bar: 5 µm. (B) There was a ∼20% reduction in the number of cross-sectional myelin figures …
Figure 9
Gria3null2–/–4–/– mice lack AMPAR-mediated inputs.
(A) The kainate-evoked current is greatly reduced in OPs of P14 Gria3null2–/–4–/– mice compared to Gria3null2–/– littermates and is less effectively blocked by the AMPAR antagonist GYKI-52466 (GYKI, …
Figure 10
Gria3null2–/–4–/– mice generate fewer OLs in the corpus callosum.
(A) CC1 immunolabelling marks differentiated OLs in Gria3null and Gria3null2–/–4–/– mice. (B) The density of CC1+ OLs is significantly less (by ∼22% at P14 and ~26% at P53) in Gria3null2–/–4–/– mice …
