Figure 1.
Expression of GAD65 and GAD67 in rat cortex (Ctx), hippocampus (HC), cerebellum (Cbl), and olfactory bulb (OB) at different postnatal ages.
Probes were examined for GAD transcript expression by qPCR. Numbers of transcripts were calculated per 1000 transcripts of GAPDH. Data represent mean±SEM of five individual animals. #P≤0.05, compared with expression of GAD67 within the same region and at the same postnatal day and as evaluated by Mann-Whitney U test. *P≤0.05, the comparison of regions at each analyzed postnatal day (upper tables) and the comparison of ages within each region (lower tables) as analysed by one-way ANOVA followed by Tukey's post hoc test.
Figure 2.
Distribution of GAD67 splice variants in rat brain at P1 (left panels) and P90 (right panels).
GAD67 mRNA (A, B) and embryonic transcripts (EGAD; C–G) were detected using radioactive in situ hybridization. Autoradiographic film images of saggital sections showed a predominant expression of GAD67 mRNA in the olfactory bulb (OB) at both ages (A, B) and a moderate expression in the cerebellum at P90 (B). Intense labeling of embryonic messages was found in the subventricular zone (SVZ; C, E) and the olfactory bulb at P1 (C). The expression persisted until adulthood [D, F, G (enlarged SVZ from emulsion autoradiography)]. Sense probe for embryonic GAD67 mRNA was used as a negative control (H, I). Black spot in H displays an artefact. CBL – cerebellum; GCL – granule cell layer; HC – hippocampus; OB – olfactory bulb; PGL – periglomerular layer; RMS – rostral migratory stream; RT – reticular thalamic nucleus; S – striatum; SEL – subependymal layer; SVZ – subventricular zone. Scale bar A–F, I, H: 2 mm; G: 100 µm.
Figure 3.
Expression of embryonic splice variants in rat cortex (Ctx), hippocampus (HC), cerebellum (Cbl), and olfactory bulb (OB) at different postnatal ages.
Probes were examined for embryonic GAD67 transcript expression by qPCR. Embryonic transcripts were jointly (EGAD) and separately (I-80 and I-86) amplified. Numbers of transcripts were calculated per 1000 transcripts of GAPDH. Data represent mean±SEM of five individual animals. *P≤0.05, the comparison of regions at each analyzed postnatal day (left table) and the comparison of ages within each region (right table) as analysed by one-way ANOVA followed by Tukey's post hoc test.
Table 1.
Sequences of gene specific primers and associated amplicon lengths and probes for in situ hybridization.