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. 2006 Aug 17;576(Pt 3):849–864. doi: 10.1113/jphysiol.2006.114702

Figure 2. Composition of HCN family proteins in the AVCN, MNTB and LSO.

Figure 2

A, all red staining represents anti HCN1, -2 or -4 labelling (see row label for subunit). Green labelling, shown in high magnification images and MNTB nucleus outline, represents presynaptic terminals as stained with anti-vesicular glutamate transporter 1 (VGlut1). The HCN1 antibody showed strong membrane labelling of somata and processes in the AVCN and the LSO but only weak staining was observed in the MNTB (by comparison, some HCN1 staining of the MSO can be seen to the left of the MNTB). The HCN2 antibody labelled all nuclei diffusely, with some somatic and some surface labelling in the AVCN. The MNTB showed the highest levels of HCN2 expression of the three nuclei. The HCN4 antibody labelled membranes of neurons in the AVCN and LSO nicely. The MNTB also expressed some HCN4 labelling that was co-localized with VGlut1, showing a probable presynaptic labelling (see inset). B, fluorescent images of brainstem slices shows strong anti-HCN1 labelling (red) in the LSO, while the AVCN (magnification of the dorsal part) have robust staining compared with the MNTB showing very weak immunoreactivity (scale bars: 50 μm (left panel), 200 μm (right panel). C, optical density measurements of HCN1 immunoreactivity within neurons in the AVCN, LSO and MNTB. Significance (**) represents P < 0.01, paired Student's t test, error bars represent s.d.