Fig. 4

Differentiation and migration of GABAergic neurons. (A–H) Expression of ptf1a:EGFP (green) and Pvalb7 (magenta) in the cerebellum at 1, 2, 3, and 5 dpf. Dorsal views (A–D), lateral views (E–H) with rostral to the left. (Ca, Da) Parasagittal sections at the levels indicated in (C) and (D). Note that many cells were positive for both ptf1a:EGFP and Pvalb7 (arrows, Ca, Da) (I, J) Parasagittal (I) and cross-sectional (J) views showing the expression of ptf1a mRNA (magenta) and ptf1a:EGFP (green) at 5 dpf. Most cells in the VZ were both ptf1a⁺ and ptf1a:EGFP⁺ (arrows). ptf1a^- ptf1a:EGFP⁺ cells found superficial to the VZ were immature neurons derived from the ptf1a⁺ neural progenitors in the VZ. (K) BrdU incorporation. Tg(ptf1a:EGFP) fish were labeled with BrdU at 5 dpf for 1 h, and proliferating cells were analyzed by immunostaining with anti-BrdU (magenta) and anti-EGFP (green) antibodies. Dorsal stacking view (K) and parasagittal section (Ka) are shown. BrdU signals were detected in ptf1a:EGFP⁺ cells in the VZ (arrows). (L) Birthdate analysis. Larvae were labeled with BrdU at 3 dpf and fixed at 7 dpf. The larvae were stained with anti-BrdU (magenta) and anti-Pvalb7 (green) antibodies. Dorsal stacking view (L) and parasagittal section (La) are shown. Note that BrdU signals were detected in Pvalb7⁺ Purkinje cells (arrows). (M) Schematic representation of development of GABAergic neurons. PCL, Purkinje cell layer; other abbreviation is described in [Fig. 1] and [Fig. 3]. Scale bars: 50 μm (Ca, J), 100 μm (K), 200 μm (A, E).