Fig. 4

Glycine signaling defects induces a drastic loss of nestin⁺ NSCs subpopulation. (A) Using nestin as a marker of NSCs subpopulation, nestin transcripts were analyzed by whole-mount in situ hybridization. A drastic loss of nestin⁺ subpopulation was observed between Ctrl-MO (i), and Glr-MO (ii) conditions at 15 and 24 hpf. (B) Double tg(GFAP:GFP; HuC:RFP) line was used to analysis nestin (white) and HuC⁺ (Bleu) expressions. Major reduction of nestin⁺ signals in Glr-MO conditions (ii) compared with Ctrl-MO (i) at 24 and 48 hpf. In addition to reduction of HuC+ signals in Glr-MO condition (ii). (C) Quantification of nestin-GFP embryos upon disruption of glycine signaling reveal a drastic reduction of GFP signal compared with control. However, co-injecting of p53-MO and Glr-MO rescue partially nestin⁺ subpopulation. For both ISH and IHC, n > 20 embryos per sample. IHC, immunohistochemistry; ISH, in situ hybridization.