|
Fig. 5 nrg1 type II signals are required in RB neurons for normal myelination in the spinal cord
(A) Illustration showing the location of the zCREST3 upstream of the islet1 gene. In transient transgenic experiments, zCREST3:dsRed shows exclusive expression in RB neurons with characteristic sensory arbors. Scale bar, 100 μm.
(B) Diagram of cell type-specific CRISPR construct design, allowing RB expression of Cas9, and ubiquitous expression of EGF or isoform-specific nrg1-targeted sgRNAs.
(C) Confocal images of oligodendrocytes (OL) in the dorsal spinal cord labeled using the claudink:GFP transgene. A dotted circle indicates the OL cell body. Scale bar, 20 μm.
(D) Quantification of total myelin sheath length produced by individual OLs in animals expressing RB-specific Cas9, with indicated nrg1 EGF and isoform-specific sgRNAs. Each point represents an individual OL; at least 30 OLs quantified from ≥3 animals per category; one representative experiment shown from three replicate experiments. Error bars represent mean ± SD. A t test was used to assess significance, and p values were adjusted for multiple comparisons. See also Figures S4, S5, and S6.