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Fig. 4. PTFla is required for generation of dorsal horn GABAergic neurons. Transverse sections through spinal cord cervical regions of Ptf1a^Cre/+ and Ptf1a^Cre/Cre mouse E16.5 embryos were processed for mRNA in-situ hybridization with the GABAergic marker gene Gad1 (A,B).Note the complete absence of Gad1 in the dorsal regions in the absence of Ptf1a. Loss of ventral Gad1 was not consistently observed. (C-H) Anti-GFP antibody was used to detect YFP in Ptf1a^Cre;R26R-stop-YFP E16.5 embryos. YFP acts as a lineage marker for cells that have expressed the Ptf1a locus. (C) The lineage reporter YFP is detected largely in the dorsal regions in wild type. (D) By contrast, in the mutant, YFP is detected at higher levels in more cells and with a different organization (arrowheads). The boxed area in C is the approximate area shown in E-H. Immunofluorescence for GAD67 (E) and the neurotransmitter GABA (G) co-localize with YFP in embryos heterozygous for Ptf1a (arrows) but not in embryos lacking Ptf1a (F,H). Panels E-H are all from dorsal horn regions. Scale bar: 110 µm in A,B; 13 µm in E-H.

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