Figure 3

Co-distribution of EGFP and glial markers in the basal hypothalamus.

(A–R) Rat frontal sections of the hypothalamus (40 μm) were analyzed using anti-vimentin (red) and anti-GFAP (red) antibodies as well as EGFP fluorescence (green) in cells transduced with AdshMCT1 at 96 h post-injection and the Hoechst nuclear stain (white). (A–D) Low magnification for EGFP (A,B) green, GFAP (C) red and vimentin (D) red expression in α-tanycytes. (E,F) 3D reconstruction of EGFP co-distribution with vimentin (E) and GFAP (F) in α-tanycytes. (G,H) One focal plane of the high magnification images of α-tanycytes for EGFP co-distribution with anti-vimentin (G) and anti-GFAP (H) antibodies. EGFP expression is observed in cells lining the 3V, co-localizing with vimentin (G) arrowheads and is not detected in GFAP-positive cells (H) arrows. (I–L) EGFP expression in β-tanycytes (I–J) green, GFAP (K) red and vimentin (L) red. (M,N) 3D reconstruction of EGFP co-distribution with vimentin (M) and GFAP (N) in β-tanycytes. (O,P) One focal plane of the high magnification images of β1-tanycytes (O) and β2-tanycytes (P) for EGFP co-distribution with vimentin. (Q,R) One focal plane of the high magnification images of β1-tanycytes (Q) and β2-tanycytes (R) for EGFP co-distribution with anti-GFAP. EGFP was expressed in both β1 and β2-tanycytes, co-localizing with vimentin (O,P) arrowheads and not with GFAP-positive cells (Q,R) arrows. α, α-tanycytes; β1, β1-tanycytes; β2, β-tanycytes; 3V, third ventricle; ME, median eminence. Scale bar: 150 μm.