Figure 1

Hepcidin mRNA expression analysed by RT-PCR and in situ hybridisation. A: Analysis of hepcidin mRNA expression by RT-PCR in adult rat brain (n = 3). Gel banding pattern was - sub ventricular zone (SVZ), olfactory bulb (OB), frontal cortex (FCx), hippocampus (HC), dentate gyrus (DG), corpus callosum (CC), cerebellum (CB) amygdala (AMD), thalamus (TH), choroid plexus (CP) and brain stem (BS). GAPDH was used as loading control. B: Graph shows the percentage of image grayscale intensity above background (mean of three samples). Statistical significance compared to whole brain control (dashed line). * = p < 0.05, ** = p < 0.01, *** = p < 0.005. Hepcidin mRNA expression was analysed by in situ hybridisation using an antisense DIG-labelled hepcidin probe. mRNA was not visible in cortex (C), and only seen in blood vessels, and choroid plexus (D and E, localisation of hepcidin mRNA indicated by arrows). To detect low-abundant mRNA an antisense radioactively labelled ([³⁵S]-dATP) probe was applied to coronal sections of rat cortex, hippocampus and dentate gyrus (F and G). Hepcidin mRNA was not detectable in cortical neurons. Probe activity was confirmed by the finding of a strong signal using an antisense β-actin probe applied to a section of rat cerebellum (H). Hepcidin mRNA was restricted to choroid plexus when the radioactive probe was applied to a section of human brain (I, indicated by arrows). A strong signal was detected on a section of rat liver included as a positive control (J). The scale bar in C represents 50 μm in G; C to F = 100 μm; 25 μm in H and J, 70 μm in I.