Figure 9

RNA in situ hybridization of gad1b and double RNA in situ hybridization for slc2a3a and gad1b, a marker for GABAergic neurons. For comparison, a single in situ hybridization for gad1b was performed at 24 hpf (A,B,B’) and 36 hpf (C,D,D’) old zebrafish embryos. Boxes in (A,B,B’,C,D,D’) depict location of high magnification pictures a₁,a₂,b₁,b₂ and c₁, c₂, d₁, d₂, respectively. In addition, a 24 hpf old zebrafish embryo was stained with slc2a3a DIG in situ probe (blue) and gad1b FLUO probe (red). Upper rows show lateral views (A,C,E) and lower rows ventral views (B,B’,D,D’,F). Anterior is to the left. Boxes in (E,F) depict location of high magnification pictures e₁–e₄ and f₁–f₄, respectively. Boxes in e₁ and e₂ and f₁, f₂, and f₄) represent higher magnification pictures shown in e₁’, e₂’ and f₁’, f₂’, and f₄’. Box in e₁’ represents location of even higher magnification shown in e₁”. White arrows mark double-positive cells or cell populations. Red arrow marks an example of cells only positive for gad1b and blue arrows cells only positive for slc2a3a. Scale bars in (A,E), 100 μm and pertains to (A–F); scale bar in a₁, 50 μm and valid for a₁–d₂; scale bar in e₁, 50 μm and pertains to e₁–e₂ and f_1–2; scale bar in e₃, 50 μm and valid for e₃-e₄, and f_3–4, scale bars in e₁’–f₄’, e₁’, 10 μm.