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. Author manuscript; available in PMC: 2022 Mar 23.
Published in final edited form as: Nat Neurosci. 2022 Mar 3;25(3):280–284. doi: 10.1038/s41593-022-01023-7

Fig. 2. Early OPC depletion causes formation of aberrant RGC arborizations.

Fig. 2

a) Time-lapse showing dynamic interactions between RGC axon arbors and OPC processes. Dashed box indicates position of timelapse shown. Arrows indicate when extending RGC process interdigitates with OPC process and subsequently retracts (see Supplementary Video 6 for 3D rotation to demonstrate contact). Pie chart shows frequency of RGC retractions with and without prior OPC contact. Scale bars: 10 μm (top), 2μm (bottom).

b)Timelines of manipulations in this figure.

c and d) Example images of NTR-mediated OPC ablation (c) and olig2 morpholino-mediated OPC depletion. Dashed lines indicate neuropil. Scale bars: 25μm.

e and f) Increased formation of ectopic RGC axon branches extending outside tectal neuropil (arrows) upon early ablation of OPCs (mean 7.5±5.1 S.D. in control vs. 22.0±5.3 in OPC-NTR ablation, n=14/15 animals from four experiments, unpaired two-tailed t test, t=7.466, d.f.=27)._Dashed lines indicate border between NP and PVZ. Scale bar: 20 μm.

g) Increased formation of ectopic RGC axon branches upon genetic OPC reduction (olig2 morphants) but not upon microglial depletion (irf8 morphants) (mean 7.9±4.3 S.D. in control vs. I6.2±3.9 in olig2 MO vs. 9.7±4.6 in irf8 MO, n=21/25/20 animals from three experiments, one-way ANOVA, F(2, 63)=23.69).

h and i) Increased size of single RGC arbors upon early OPC ablation (top) whilst maintaining single lamina layering (bottom) (median 287±403/235 I.Q.R. in control vs. 393±46l/286 in OPC-NTR ablation, n=26/27 axons in 24/23 animals from three experiments, two-tailed Mann-Whitney U test, U=210). Scale bars: 10 μm.

j) Increased size of single RGC arbors upon genetic OPC depletion (olig2 morphants) but not upon microglial depletion (irf8 morphants) (median 304±39l/255 I.Q.R. in control vs. 355±458/326 in olig2 MO vs. 285±323/229 in irf8 MO, n=21/30/16 axons in 11/14/12 animals from five experiments, Kruskal Wallis test, test statistic=9.9)