Figure 3.

RAGE and ALCAM are expressed on non-neuronal cell populations. (A) CML binds to proteins of RAGE (molecular weight ≈ 75 KDa) and ALCAM (molecular weight ≈ 105 KDa); the vehicle does not bind to protein of either receptor, as illustrated by no band detected in binding of ALCAM. (B) RAGE and ALCAM gene expression in mediobasal hypothalami of chow or HCHF mice (n = 6 for chow or for HCHF, P = 0.019 for RAGE, P = 0.006 for ALCAM). (C) RAGE is intensely expressed by microglia (iba1-ir, indicated by white arrowheads). Higher magnifications of the areas framed by dashed lines are presented in D. (E) RAGE is intensely expressed on endothelial cells (laminin-ir, indicated by white arrows). Higher magnifications of the areas framed by dashed lines are presented in F. (G) CML stimulates TNFα, but not PDGF-B, gene expression in cultured primary microglia (n = 6 wells of cells for vehicle, n = 5 for TNFα treatments, P = 0.04 for TNFα). (H & I) CML stimulates microglial reactivity in the mediobasal hypothalamic area, arrowheads point to the areas where the tip of the infusion probes located. (J) Iba1-ir cell number and cell coverage in H & I (n = 4 mice for vehicle, n = 5 for CML). (K) ALCAM is expressed on part of the vasculature (laminin-ir, indicated by white arrows, two pericytes are indicated by white arrowheads); higher magnifications of the areas framed by dashed lines are presented in L. (M) ALCAM is expressed on pericytes (PDGFRβ-ir, white arrowheads). Higher magnifications of the areas framed by dashed lines are presented in N. Scale bar: 30 μm in C, E, K and M, 7.5um in D, F, L and N. Data are presented as means ± s.e.m. *P < 0.05, **P < 0.01. P values for unpaired comparisons were analyzed by two-tailed Student's t test.