In vitro binding experiments were performed as previously described [9] with 20-μm brain cryosections from transgenic APP23 (N = 2), Tg2576 (N = 2) and APPswe-PS1dE9 (N = 2) mice and wild-type control mice (N = 5) that were sacrificed after in vivo studies were finalized (Table 2).
For isolating proteins from hindbrain of 4%-PFA fixed brain cryosections from wildtype and Sim1-/ mutants, the method described by Scicchitano et al., [42] has been applied, using the Qproteome FFPE Tissue Kit from Qiagen, following the manufacturer's instructions.
We co-stained brain cryosections from 7-day-old rat pups with antibodies against VPAC2 and a ciliary marker, Arl13B (Caspary et al., 2007) and observed that the anti-VPAC2 antibody stained cilia in various brain regions including the thalamus and SCN (Fig. 2C,D).
Furthermore, we detected VPAC2-positive cilia in Aldh1L1+ cells from brain cryosections of 7-day-old rat pups (Fig. 3D).
Histopathologically altered areas of hippocampus CA1, containing birefringent material, and ultrastructurally packed with fibrils from brain cryosections of a 6-month BMAA rat treated neonatally were isolated using laser capture microdissection.
Table 2 Summary of the brain tissue used for in vitro experiments APP23 Tg2576 APPswe-PS1dE9 TG (mo) WT (mo) TG (mo) WT (mo) TG (mo) WT (mo) In vitro ARG 18, 21 27 27 22 19 19 Aβ1-40 21, 27 24, 27 22 22 17, 19 19 Thioflavin S 21, 27 27, 27 22 22 19, 19 19 Ages of the transgenic (TG) and wild-type (WT) mice from which the brain cryosections for stainings in this study were obtained.
Twenty-micrometer coronal brain cryosections were cut serially so to have sections distanced by ≈1 mm and stained with cresyl violet [21].
Ten-µm brain cryosections were prepared, fixed in 75% ethanol for 30s.
Radioactive in situ hybridization was performed on 16 µm brain cryosections as described previously [88], [89].
An additional colocalization experiment, also performed on adult brain cryosections, revealed strong β-gal labeling in cells expressing the Nr2f2 mouse gene in brain regions extending from the PMCo to the MePV.
