Fig.6.

Staining of Aβ+ or Aβ– treated brains with Aβ-N and Congo red (A), microglial marker Iba1 (B) in Aβ+ or Aβ– treated mouse brains, quantification of Aβ burden (C–G), Iba1 (H), western blot of AβPP and CTFs (I), ELISA of sAβPPα (J), sAβPPβ (K), western blotting of total tau (L), phosphorylated tau (M), CSF Aβ₄₀ (N), and Aβ₄₂ (O) measured by ELISA. A) Aβ-immunoreactive load using Aβ-N was suppressed in the Aβ+ treated group compared with that of the Aβ– group. Most Aβ staining showed diffuse plaques that are not stained with Congo red. B) Microgliosis was weaker in the Aβ+ treated group than that in the Aβ– treated group. Bar represents 100 μm in A and 200 μm in B. C) The area labeled by Aβ-N was significantly suppressed in the Aβ+ treated group compared with that in the Aβ– treated group (p < 0.0001). D, E) The area occupied by diffuse plaques was more than 3-fold of that by core plaques. Both plaques were suppressed by Aβ+ treatment (D, p < 0.0001; E, p < 0.0001). Large part of the Aβ decrease by immunization consisted of diffuse plaques. Aβ burdens detected by anti-Aβ₄₀ (F) and anti-Aβ₄₂ (G) were significantly decreased in the Aβ+ treated group (F, p < 0.001; G, p < 0.0001). H) The area of Iba1 staining in Aβ+ treated mice was smaller than that in Aβ– treated mice (p < 0.0001). The numbers of mice analyzed: 23 weeks (n = 9 for Aβ+, n = 11 for Aβ–), 43 weeks (n = 7 for Aβ+, n = 6 for Aβ–), and 59 weeks (n = 6 for Aβ+, n = 7 for Aβ–). I) AβPP, CTFβ, and CTFα, and other CTFs in SDS fractions did not differ between the Aβ+ and Aβ– treated groups from 23 to 59 weeks. (J, K) The amount of sAβPPα and sAβPPβ in the TBS fractions detected by ELISA did not differ between Aβ+ (Red) and Aβ– (Blue) treated groups from 23 to 59 weeks. L, M) The amount of total tau (TAU-5) and phosphorylated tau (pTau) (PHF-1) did not differ between Aβ+ and Aβ– treated mice from 23 to 59 weeks. N, O) Aβ₄₀ and Aβ₄₂ in CSF decreased with aging. They were decreased in Aβ+ treated mice compared to Aβ– treated mice, although there was no significance difference between Aβ+ and Aβ– mice (Aβ+ n– =– 7 for 23 weeks, n = 4 for 43 weeks and n = 6 for 59 weeks; Aβ– n = 9 for 23 weeks, n = 2 for 43 weeks, and n = 4 for 59 weeks).