Figure 4. GABAergic interneuron loss in the P2 hippocampus and dentate gyrus of Evf2TS/TS mutant mice.
a. RNA in situ hybridization analysis of Evf2 expression in the subventricular zone and cells lining the lateral ventricle as they migrate to the hippocampus and dentate gyrus. scale bar=130μm; Or, oriens; Py, pyramidal; Rad, radiatum; LMol, lacunosum molecular; Mol, molecular layer of the dentate gyrus; DGC, dentate granule cell layer; DG, dentate gyrus; LV, lateral ventricle. b. Quantification of the number of GAD67-expressing GABAergic interneurons in the dentate gyrus (DG), and hippocampal CA1 and CA3 regions. Error bars: S.E.M. Statistical analysis: Mann Whitney U-test, p =.025 for DG, CA3, CA1; n= 3 for wt, n=3 for Evf2TS/TS. c–d. RNA in situ hybridization of GAD67 expressing interneurons in the hippocampus, scale bar=135μm e–f. Immunohistochemistry, anti-GABA antibodies (green) and DAPI (blue nuclei), scale bar= 200 μm. g–h. RNA in situ hybridization of v-glut1 expressing neurons in the hippocampal CA3 region shows similar expression in wt and Evf2TS/TS, scale bar=80μm. i–j. Immunohistochemistry using the apoptosis TUNEL assay (green) and DAPI (blue nuclei) in the hippocampal CA3 region, scale bar= 10μm. k. Quantification of the number of TUNEL-positive cells in the dentate gyrus (DG), and hippocampal CA1 and CA3 regions shows no difference in cell death between wt and Evf2TS/TS mutants. Error bars: S.E.M. Statistical analysis: Student’s t-test, p >0.05 for DG, CA3, CA1; n= 3 for wt, n=3 for Evf2TS/TS. * p < 0.05; t-test or Mann Whitney U-test