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Fig. 1. Expression and inactivation of COUP-TFI in the developing forebrain. (A-J) Coronal sections immunostained for COUP-TFI at the ages indicated above. A to D are consecutive rostral to caudal sections. (E) Enlargement of the presumptive cingulate cortex in which COUP-TFI is present in scattered cells (arrows). (F) Detail of one half of the cingulate cortex, double immunolabelled for COUP-TFI (green) and Reelin (red), shows that COUP-TFI-positive cells do not co-localize with the marginal zone marker Reelin. (G-J) Note how COUP-TFI starts to be expressed in the dorsal thalamus (dTh) from the onset of thalamic differentiation and is maintained at later stages in distinct nuclei. (I) In the hippocampus high levels of COUP-TFI are present in the hippocampal plate and in the intermediate zone. Scattered cells are also detected in the inner marginal zone. (J) Detail of the dorsal thalamic region in a section posterior to (I) shows high expression of COUP-TFI in the lateral geniculate and ventroposterior nuclei. Dashed lines demarcate the LGN and VP thalamic nuclei. The zona incerta is also positive for COUP-TFI. (K) Diagram showing the gene targeting strategy used to obtain the COUP-TFI^null allele after Cre-recombination in embryonal stem cells. The lox P sites are depicted as triangles. Blue boxes represent exons. Appropriate restriction enzyme sites are indicated. Red lines indicate the expected fragment size after EcoRI digestion of COUP-TFI^wt, COUP-TFI^floxneo and COUP-TFI^null genomic DNA, as shown in a Southern blot (L). (M) PCR genotyping on wild-type (+/+), heterozygous (+/-) and homozygous (-/-) mice using the primers indicated in K as black arrows. (N) Western blot shows total absence of COUP-TFI protein in homozygous mutant animals. (O) Coronal sections of wild-type and homozygous E15.5 embryos confirm complete absence of COUP-TFI protein in homozygous brains. Scale bars: 200 µm. aa, anterior amydgaloid area; aon, anterior olfactory nucleus; BG, basal ganglia; cam, claustro-amygdaloid complex; cp, cortical plate; hip, hippocampus; hp, hippocampal plate; ic, internal capsule; imz, inner marginal zone; iz, intermediate zone; LGN, lateral geniculate nuclei; pir, piriform cortex; rms, radial migratory stream; s, septum; sp, subplate; VP, ventroposterior nuclei; ZI, zona incerta.

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