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). Glast-(h, Some ependymal cells showed low levels of vimentin in their soma (a-d, empty arrowheads) but prominent expression in their basal processes, some of which co-expressed nestin (crossed arrows in b-d). Ependymal cells were heterogeneous for the expression of GFAP, with GFAP-negative (q, empty arrowhead) next to GFAPpositive cells (q, plain arrowhead). Note the strong expression of GLAST (h, j) and GFAP (o, q) in the hypocellular gap. Arrows point to ribbon cells that co-express nestin, Sox2 and vimentin, GLAST or GFAP. The same cells are pointed in main panels (a, h, o) and highmagnification views. Empty arrows in main panels point to vimentin-(a), GLAST-(h) and GFAP-expressing cells (o) in the parenchyma behind the ribbon, Expression of NPC markers in ependymal and ribbon cells of the human third ventricle. Nestin (green) and Sox2 (magenta) were co-immunodetected with vimentin

G. and H. Gap,

, Figure 5. Expression of NPC markers in ventricular protrusions of the human third ventricle

, These structures contain nestin (a-i, green)/Sox2 (magenta)-immunoreactive cells that co-express vimentin (a-c, j-l), GLAST (d-f) and GFAP (g-i) (arrows, co-expressing cells are shown at higher magnification in insets). The subependymal region and buds are devoid of NeuN-immunoreactive cells, Protrusions of various sizes and forms are found all along the third ventricle

, Expression of NPC markers in the median eminence of the human hypothalamus. a: Nestin (green) and Sox2 (magenta) are expressed in tanycytes, which line the floor of the third ventricle. Crossed arrows point to tanycyte nuclei and arrows show tanycyte processes. b-d: co-immunodetection of nestin (green) and vimentin (b, white)

). Gfap-(d,-white, The three panels below each main panel show a high magnification view of the boxed area. Nestin-immunoreactive tanycyte processes co-express vimentin

, White arrowheads point to nestin-positive processes devoid of vimentin (b), GLAST (c) or GFAP (d) immunoreactivity. Vimentin-positive (b, empty arrowhead) and GFAP-positive (d, empty arrowhead) processes devoid of nestin immunoreactivity were also seen

. 3v, . Me, and . Eminence, Scale bars = 50 µm in a, 20 µm in other panels. Figure 7. Expression of NPC markers in the human suprachiasmatic nucleus. a: Immunofluorescent staining of VIP (white) was performed to ascertain the correct localization of the suprachiasmatic nucleus on human hypothalamus sections. Crossed arrows point to VIP-immunoreactive neurons

, The inset is a high-magnification view of the boxed area in the main panel

, Note the co-existence of small process-bearing nestin/Sox2 co-expressing cells (arrows) with

, Arrows point to triple labelled cells and arrowheads show nestin-negative/Sox2-positive cells. l, m: Co-immunodetection of GLAST (green), Sox2 (magenta) and NeuN (white) in the human suprachiasmatic nucleus. Empty arrows point to NeuN-positive neurons that lack GLAST and Sox2 expression. The arrow shows a GLAST/Sox2 co-expressing cell devoid of NeuN, Sox2-positive/nestin-negative cells (arrowheads). c-k: triple immunofluorescent stainings of nestin (green), Sox2 (magenta) and vimentin (white, d, e)

, The 2 cells pointed by the arrows in the main panel (a) are shown at higher magnification in the panels b-e. Very rare nestin/Sox2-co-expressing cells were immunoreactive for Ki67 (arrows). The empty arrow, Co-immunofluorescent staining of nestin (green)

, CD68 (green) reveals the presence of double-labeled cells (arrow) in addition to Ki67-positive/CD68-negative cells (plain arrowheads) and Ki67-negative/CD68-positive cells (empty arrowhead), Arrowheads in b-e point to Ki67-positive cells that do not co-express nestin and Sox2. f-h: Co-immunofluorescent staining of Ki67 (white) and

, Representative photographs were taken at the level of the tuberal region of the hypothalamus. The rat third ventricle is shown as a whole while the third ventricle from the mouse and grey mouse lemur is shown in its most dorsal (a-d, l-n) and ventral (e-h, o-q) portions. Co-immunostainings of Sox2 (magenta, a, e, i, l, o), nestin (green, b, f, j) and vimentin (white, c, g, inset in j, m, p), Expression of NPC markers along the third ventricle of the mouse (a-h), rat (i-k) and grey mouse lemur (l-q)

G. Pellegrino, C. Trubert, J. Terrien, F. Pifferi, D. Leroy et al., in ependymal cells (arrowheads) and is enriched in the gap and ribbon. The ribbon contains a population of cells that co-express nestin, Sox2 and GLAST (arrows, j-l). m-p: GFAP (white) is enriched in the gap and ribbon. The ribbon contains a population of cells that coexpress nestin, Sox2 and GFAP (arrows, n-p). GFAP is also expressed in parenchymal astrocytes (crossed arrows in m) but is absent from most ependymal cells (empty arrowheads). Rare ependymal cells express GFAP (plain arrowhead, Nuclei were counterstained with Hoechst (blue). Microphotographs representative of the ventral, vol.526, pp.1419-1443, 2018.

, Arrows point to nestin/Sox2 co-expressing cells; the arrowhead points to Sox2-positive/nestin-negative ependymal cells. b: Immunodetection of GFAP (white) reveals the presence of numerous astrocytes (arrows). c: Vimentin immunoreactivity (white) was detected in capillaries (asterisks) and small stellate cells (arrows), Expression of NPC markers in the dentate gyrus of the human hippocampus