The levels of different Aβ isoforms in the brain detergent lysates of experimental mice studied by Sandwich ELISA.

<p><b>A</b>—total Aβ₄₀ and Aβ₄₂, <b>B</b>—Aβ₄₀ and Aβ₄₂ bound to α7 nAChR in mice immunized with 7(1–208) (n = 8) or injected with LPS (n = 5) compared to non-treated animals (Ctrl, n = 9) and to those “immunized” with complete Freund’s adjuvant (CFA, n = 5). The columns correspond to M±SE (n = 5); *—p<0.05; **—p<0.005; ***—p<0.0005 compared to Ctrl.</p

The level of α7, β2 or β4 nAChR subunits in the brain sections of experimental mice studied by immunohistochemistry.

<p>Confocal microscopy images of the hippocampus CA1 and striatum of non-treated (Ctrl), α7(1–208)-immunized or LPS-injected mice stained with biotinylated α7-, β2- or β4-specific antibodies and developed with Extravidin-Cy3 (<i>red</i>). Cell nuclei are stained with DAPI (<i>blue</i>). Bar corresponds to 50μm, actual for each fragment of the panel.</p

Episodic memory of experimental mice studied in the “Novel Object Recognition” test.

<p>Discrimination indexes calculated for mice immunized with 7(1–208) (n = 8) or injected with LPS (n = 5) compared to non-treated animals (n = 9) or those “immunized” with complete Freund’s adjuvant (CFA, n = 5). ***—p<0.0005 compared to non-treated mice (Ctrl).</p

The GFAP-positive astrocytes (A) and the number of nucleated cells (B) in the brain sections of experimental mice studied by immunohistochemistry.

<p><b>A</b>—Confocal microscopy images of the hippocampus CA1 (Hip), motor/somatosensory cortex (Crtx) or striatum (Str) of non-treated (Ctrl), α7(1–208)-immunized or LPS-injected mice stained with rabbit GFAP-specific antibody and developed with anti-rabbit Alexa 488 (<i>green</i>). Cell nuclei are stained with DAPI (<i>blue</i>). Bar corresponds to 50μm, actual for each fragment of the panel. <b>B</b>—The number of nucleated cells (DAPI-positive) in corresponding brain regions studied in all available sections (12 to 16 for each treatment for each region); *—p<0.05; **—p< 0.005.</p

The level of Aβ₄₀ and Aβ₄₂ in the brain sections of experimental mice studied by immunohistochemistry.

<p>Confocal microscopy images of the hippocampus CA1 or motor/somatosensory cortex of non-treated (Ctrl), α7(1–208)-immunized or LPS-injected mice stained with biotinylated Aβ₄₀- or Aβ₄₂-specific antibodies and developed with Extravidin-Cy3 (<i>red</i>) and with α7-specific antibody developed with anti-rabbit Alexa 488 (<i>green</i>). Bar corresponds to 50μm, actual for each fragment of the panel.</p

Visualization of biotinylated antibody within the brain at different periods after injection.

<p>Confocal microscopy images of the striatum of mice injected with α7(1–208)-specific antibody (A-B) or non-specific IgG (C) in 15 min (A) or 3h (B-C) after injection. Antibodies were developed with Extravidin-Cy3 (<i>red</i>). Bar corresponds to 100μm, actual for each fragment of the panel. D—Antibody-specific ELISA signal of the primary brain supernatants and detergent lysates of mice either pre-treated or not with LPS; the brains were removed in 15 min or 3 h after the antibody injection. Each column corresponds to mean±SE of three repeats in ELISA; *—p<0.05; **—p<0.005 compared to the data of LPS non-treated mice.</p

The level of nAChR-specific antibodies in the blood and of nAChR subtypes in the brain of experimental mice studied by ELISA (A) or Sandwich ELISA (B).

<p><b>A</b>—7(1–208)-specific antibodies in the blood sera (1:50) of mice immunized with 7(1–208) (n = 8) or injected with LPS (n = 5) for 5 months compared to non-treated (Ctrl, n = 9) and adjuvant-“immunized” animals (CFA, n = 5). <b>B</b>—3, 4, 7, β2 and β4 nAChR subunits in the brain detergent lysates of the same groups of mice (4 mice from each group). <b>C</b>—Pearson coefficients (R) of correlation between the levels of nAChR subunits in the brain and those of 7(1–208)-specific antibodies in the blood. The columns correspond to M±SE, *—p<0.05; **—p<0.005; ***—p<0.0005 compared to Ctrl.</p