Figure 4

MTH is necessary for developing and correctly positioning neural cells in the spinal cord. WISH expression pattern of (A)her2, (B)neurog1, (C)fabp7a, (D)slc1a2b, (E)olig2, (F) IHC for Nkx6.1 in the spinal cord (SC) of zebrafish during embryonic development in MCT8MO and CTRLMO embryos. (A) Green asterisk in 25hpf MCT8MO represents absence of her2 expression. (B) Green asterisk in 32hpf MCT8MO highlights absence of neurog1 expression. (C) Green arrowheads at 32hpf CTRLMO, indicate the dorsal fabp7a+ cells which are lost in MCT8MO. Red arrowheads indicate increased fabp7a staining in the ventral domain in MCT8MO. At 48hpf, green arrowheads indicate fabp7a+ cells in the CTRLMO SC dorsal domain, which are less evident in MCT8MO. (D) At 25hpf, green arrowheads indicate misplaced slc1a2b+ cells in the ventral SC of MCT8MO. At 32hpf, the expression of slc1a2b is less abundant in MCT8MO. At 48hpf MCT8MO display reduced slc1a2b signal in cells at the most ventral and dorsal regions of the neurocoelom. The green arrowhead in the CTRLMO inset indicates an area where slc1a2b expression is less abundant in MCT8MO. (E) In CTRLMO, olig2+ cells were present in the most ventral region of the SC. At 25hpf, the green arrowhead indicates the position of olig2+ cell clusters, and the asterisk denotes the absence of cells in MCT8MO. (F) Nkx6.1 immunofluorescence detection at 20, 25, and 32hpf. A minimum of 10 individuals/conditions/gene or protein were analyzed. Images represent a lateral view of the SC (unless specified) between somites 8-12 with rostral orientation located to the left; Green dashed line represents the ventral limit of the SC; Scale bars represent 50 µm. Insets in (B–D) at 48hpf are transverse sections; Green dashed line represents the outermost boundary of the SC; Scale bars represent 20 µm.