Fig. 7.

Detection of Gal₁-mCherry immunofluorescence in brain. (A–B) In low-power micrographs (2.5 ×), specific immunofluorescence is not detected in adult wild-type (wt) brain (A), whereas in a similar section of adult GalR1-mCherry knock-in brain, high levels of Gal₁-mCherry immunoreactivity were detected (B; see F-H for same section at higher magnifications). (C–I) Higher magnification images (5 ×) show high immunoreactivity in: (C) intermediate and ventral parts of lateral septal nuclei (LSI, LSV) and scattered cells within caudate putamen (CPu), but not in the dorsal part of the lateral septal nucleus (LSD) at this level or in medial septal nucleus (MS); (D) LSI, and scattered cells within CPu; (E–F) paraventricular-, medial mediodorsal-, central medial-, paracentral-, central lateral- and intermediodorsal-thalamic nuclei (PV; MDM; CM; PC; CL; IMD), but not in dentate gyrus (DG) or medial habenula (MHb); (G) dorsomedial and ventromedial hypothalamic nuclei (DM; VMH), median eminence (ME; see below) and scattered cells within posterior and lateral hypothalamic areas (PH; LH); (H) posterior basomedial and posterodorsal medial amygdala nuclei (BMP; MePD) and LH; (I) locus coeruleus (LC) and posterodorsal tegmental nucleus (PDTg), but not cerebellum (CB). (J–M) Confocal images at higher magnification (J–L: 40 ×; M: 40 ×, zoom 3 ×; scale bars 40 μm) showing: (J) somatic cell membrane localization in some neurons within the intermediate part of the lateral septal nucleus (LSI; arrows; area boxed in (D)); (K) high levels of expression in median eminence (ME) shown either over-bright to show location of third ventricle (3V; left pair of arrows showing join of torn tissue; 30 z-sections) or less-bright image (L; 3 z-sections), with insert of boxed area magnified to show immunoreactive fibres and an absence of immunoreactive cell bodies (40 ×, zoom 6 ×; scale bar 15 μm; 3 z-sections); and (M) fibres within the locus coeruleus (LC) area, together with a single cell body (arrow; 20 z-sections), with inserts showing somatic cell membrane localization (single z-sections). Other abbreviations: aca, anterior part of anterior commissure; and 4V, 4th ventricle. Images (A–B) and (C–I) are each at the same brightness intensity, with scale bars of 1 mm (A–B, 2.5 × objective) or 0.5 mm (C–I, 5 × objective lens). Sections are similar to mouse brain atlas Figures 46 (A, B), 23 (C), 29 (D, J), 40 (E), 46 (F–H, K, L) and 79 (I, M) of Franklin and Paxinos (1997).