The loss of tamalin and Arf6 synergistically induces neuronal and oligodendrocyte degeneration during CNS development. (a–d) The representative images are transverse sections of the spinal cord at 3 dpf. The dorsal side is displayed at the top. Wildtype embryos injected with control MO (CTMO) (a) and Arf6 MO (b), or tamalin KO embryos injected with CTMO (c) and Arf6 MO (d) have been immunolabeled with anti-Hu (white) and anti-Sox10 (green) antibodies to detect neurons and oligodendrocyte lineage cells, respectively, and TUNEL staining (red) to detect apoptotic cell death. (d’,d”) High magnification images of the boxes in (d). (e–h) Lateral views of the spinal cord of 3 dpf Tg(mbp:egfp) zebrafish injected with CTMO (e) and Arf6 MO (f), or tamalin KO zebrafish injected with CTMO (g) and Arf6 MO (h). The anterior side is displayed on the left. (i–k) Quantification of the number of TUNEL+/Hu+ neurons (i), TUNEL+/Sox10+ oligodendrocyte lineage cells (j) and MBP+ mature oligodendrocytes (k). (**** p < 0.0001, *** p < 0.001, ** p < 0.01, * p < 0.05). Scale bars: (a–d”), 10 μm (e–h), 20 μm (a–d: n = 10 sections from five zebrafish, (e–h): each image from ten zebrafish). KO, knockout; MO, morpholino oligonucleotides; CNS, central nervous system; dpf, days post fertilization; TUNEL, terminal deoxynucleotidyl transferase dUTP nick end labeling; Sox10, Sry-related HMg-Box gene 10; Arf6, ADP-ribosylation factor 6; and MBP, myelin basic protein.
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