Figure 5: Both embBE and abolition of MGv waves induce changes in gene expression in the VPM thalamic nucleus.

(a) Scheme representing the microarray experiment. VPM nuclei were collected at P0 and P4, and the RNA was extracted and processed according to the Affymetrix GeneChip protocol. Bright field image showing a coronal slice after dissection of the VPM nucleus. (b) Scatter-plots showing significantly upregulated or downregulated transcripts (red and green circles, respectively) with a change of ≥1.5 or ≤−1.5-fold and a P value of ≤0.05 at P0 and P4. In the VPM, 106 transcripts were significantly regulated at P0 and 47 at P4, with an overall tendency towards upregulation. The top ten significantly upregulated or downregulated transcripts in the VPM are listed in red or green, respectively. (c) The expression of the Rorβ gene in VPM nucleus is upregulated by 1.9-fold at P0 in embBE mice (**P=0.008; Two-tailed Student’s t-test). Between P0 and P4, Rorβ expression is upregulated in the VPM 2.4-fold in control mice (***P<0.001; Two-tailed Student’s t-test) and 1.6-fold in embBE mice (*P=0.04; Two-tailed Student’s t-test). (d) In situ hybridization for Rorβ in coronal sections from control and embBE animals at P0 and P4. Note the stronger expression of Rorβ in the VPM area (asterisk) of embBE animals compared with the controls. (e) Quantitative real-time PCR for Rorβ transcripts in VPM neurons, in control media (n=12) or after treatment with 25 mM of KCl (n=13) in acute slices at E16.5 (**P=0.0042, Two-tailed Student’s t-test). (f) Quantitative real-time PCR for Rorβ in the VPM nucleus in control (n=11) and MGv^Kir (n=10) mice at E16.5 (*P=0.01, Mann–Whitney U-test). (g) In situ hybridization for Rorβ in coronal sections from control (n=5) and MGv^Kir (n=5) animals at P0. Graphs represent mean±s.e.m. Scale bars, 300 μm.