Journal: The Journal of comparative neurology

Article Title: Rax regulates hypothalamic tanycyte differentiation and barrier function in mice

doi: 10.1002/cne.23451

Figure Lengend Snippet: Confocal z-stack reconstruction of Rax fluorescent in situ hybridization (fISH) (magenta) and vimentin (Vim) immunohistochemistry (green) counterstained with DAPI in brain sections of adult C57BL/6 wild type mice (P45). (A–T) Rax is expressed along the anteroposterior axis of the ventral hypothalamus and co-localizes with vimentin in the ventral portion of the wall of the third ventricle (white arrow heads). Numbers in left bottom corner correspond to different Bregma points. Bregma −1.555mm (A–D) represents the anterior hypothalamus, Bregmas −1.655mm to −1.955 mm (E–P) represent the medial hypothalamus and Bregma −2.055mm (Q–T) represents the posterior hypothalamus. Scale bar: 100μm. 3V: third ventricle.

Article Snippet: When the staining was complete, 100μl of Vectashield hard-set mounting medium with DAPI (Vector laboratories) was applied to the slides and covered with a cover slip.

Techniques: In Situ Hybridization, Immunohistochemistry

Journal: The Journal of comparative neurology

Article Title: Rax regulates hypothalamic tanycyte differentiation and barrier function in mice

doi: 10.1002/cne.23451

Figure Lengend Snippet: Confocal z-stack reconstruction of double fISH for Rax (green), Gpr50 (magenta) mRNA and vimentin (Vim) immunohistochemistry (white) counterstained with DAPI (blue) in brain sections of adult C57BL/6 wild type mice (P45). Rax and Gpr50 are expressed by α1 (F–J), α2 (K–O), β1(P–T) and β2 (U–Y) tanycytes (Vim+ cells with process, white arrow heads) and they are absent from ependymal cells (A–E) (Vim+ cells without process, yellow arrow heads). Rax and Gpr50 are also expressed in the median eminence (U–W). 3V: third ventricle. Median eminence: ME. Scale bar: 20μm.

Article Snippet: When the staining was complete, 100μl of Vectashield hard-set mounting medium with DAPI (Vector laboratories) was applied to the slides and covered with a cover slip.

Techniques: Immunohistochemistry

Journal: The Journal of comparative neurology

Article Title: Rax regulates hypothalamic tanycyte differentiation and barrier function in mice

doi: 10.1002/cne.23451

Figure Lengend Snippet: Orthogonal view of Rax (green) and Gpr50 (magenta) double fISH combined with vimentin (Vim) immunohistochemistry (white) counterstained with DAPI (blue) in the median eminence of adult C57BL/6 wild type mice (P45) (A) Rax and Gpr50 mRNAs are not only expressed in β tanycytes located in the ependymal layer (EL) of the ME (white arrow heads), but also in other layers of the ME in close association with tanycyte processes (yellow arrows). This localization suggests that Rax and Gpr50 are translated in tanycyte processes. (B–K) Orthogonal view of the dashed square area showing a Rax+, Gpr50+ nucleus in close association with Vim staining. These Rax+, Gpr50+ cells might correspond to “astrocytic tanycytes” EL, Ependymal layer, SE, subependymal layer, FL, fiber layer, RL, reticular layer, PL, palisade layer. 3V: third ventricle. Scale bar: 20 μm.

Article Snippet: When the staining was complete, 100μl of Vectashield hard-set mounting medium with DAPI (Vector laboratories) was applied to the slides and covered with a cover slip.

Techniques: Immunohistochemistry, Staining

Journal: The Journal of comparative neurology

Article Title: Rax regulates hypothalamic tanycyte differentiation and barrier function in mice

doi: 10.1002/cne.23451

Figure Lengend Snippet: Confocal z-stack reconstruction of Rarres2 fISH (magenta) and immunohistochemistry for detyrosinated α tubulin (G-TUB) (green) counter stained with DAPI (blue) in the anterior (A), medial (B) and posterior (C) hypothalamus of adult C57BL/6 wild type mice (P45). (A–C) Top panels show that G-TUB is expressed in the dorsal region of the wall of the third ventricle along the anteroposterior axis similar to Rarres2 expression (white arrow heads). Scale bar: 100μm. Bottom panels (D–K) show an orthogonal view of the box in (A) where G-TUB (D, E) and Rarres2 (F, G) signal is observed in ependymal cells identified by the expression of G-TUB in their multiple cilia projecting to the third ventricle (green). Rarres2 signal is underneath G-TUB+ multiple cilia (J–L). G-TUB signal in hypothalamic parenchyma corresponds to primary cilia. 3V: third ventricle. Scale bar: 20 μm.

Article Snippet: When the staining was complete, 100μl of Vectashield hard-set mounting medium with DAPI (Vector laboratories) was applied to the slides and covered with a cover slip.

Techniques: Immunohistochemistry, Staining, Expressing

Journal: The Journal of comparative neurology

Article Title: Rax regulates hypothalamic tanycyte differentiation and barrier function in mice

doi: 10.1002/cne.23451

Figure Lengend Snippet: Confocal z-stack reconstruction of Rax fISH (magenta) and vimentin (Vim) immunohistochemistry (green) counterstained with DAPI (blue) in the α2 tanycytic zone of the medial hypothalamus (Bregma −1.655mm). Rax mRNA signal is reduced in Rax heterozygotes mice (Rax+/−) (E) compared to Rax wild type mice (Rax+/+) (A). (I) Digital quantification of Rax fISH signal in the α2 tanycytic zone using a three dimensional reconstruction of the signal with the spot tool of the Imaris software Unpaired t-test (n=3), p=0.01. 3V: third ventricle. Scale bar 20μm.

Article Snippet: When the staining was complete, 100μl of Vectashield hard-set mounting medium with DAPI (Vector laboratories) was applied to the slides and covered with a cover slip.

Techniques: Immunohistochemistry, Software

Journal: The Journal of comparative neurology

Article Title: Rax regulates hypothalamic tanycyte differentiation and barrier function in mice

doi: 10.1002/cne.23451

Figure Lengend Snippet: (A–H) Confocal z-stack reconstruction of DAPI staining along the third ventricular wall of the medial hypothalamus (dotted area) in adult mice (P45). (I–P) Digital three-dimensional reconstruction of the dotted areas in (A). (Q) Digital volume quantification of the wall of the third ventricle using the cell tool of the Imaris software. Rax heterozygotes have reduced volume of the ventricular wall at the ependymal zone (B, J, Q) as well as at the α2 (F, N, Q) and β1 (H, P, Q) tanycytic zones. Unpaired t-test (n=3), β1 tanycytes p=0.01, α2 tanycytes p=0.03, α1 tanycytes p=0.08, ependymal cells p=0.02. 3V: third ventricle. Scale bar: 50μm.

Article Snippet: When the staining was complete, 100μl of Vectashield hard-set mounting medium with DAPI (Vector laboratories) was applied to the slides and covered with a cover slip.

Techniques: Staining, Software

Journal: The Journal of comparative neurology

Article Title: Rax regulates hypothalamic tanycyte differentiation and barrier function in mice

doi: 10.1002/cne.23451

Figure Lengend Snippet: (A–H) Confocal z-stack reconstruction of Rarres2 fISH (magenta) and vimentin (Vim) immunohistochemistry (green) counterstained with DAPI (blue) in adult mice (P45). Rarres2 mRNA is more abundant in the α2 ventral tanycytic zone (immediately ventral to the deflection point) of Rax+/− mice (E) compared to Rax+/+ controls (A). (I) Digital quantification of Rarres2 fISH signal in the ventral α2 tanycytic zone at three different Bregma points using a three-dimensional reconstruction of Rarres2 fISH signal with the spot tool of the Imaris software. Cumulative indicates the sum of Rarres2 signal along the three different Bregma points (from 1.655 mm to −1.855 mm). Unpaired t-test (n=3), Bregma −1.655 mm p=0.05, Cumulative p=0.04. 3V: third ventricle. Scale bar: 20μm.

Article Snippet: When the staining was complete, 100μl of Vectashield hard-set mounting medium with DAPI (Vector laboratories) was applied to the slides and covered with a cover slip.

Techniques: Immunohistochemistry, Software

Journal: The Journal of Cell Biology

Article Title: FIAT represses ATF4-mediated transcription to regulate bone mass in transgenic mice

doi: 10.1083/jcb.200412139

Figure Lengend Snippet: Normal osteoblast proliferation and survival in FIAT transgenic mice. Calvarial sections from wild-type (A and C) or transgenic (B and D) animals were stained for PCNA (A and B) and with a DAPI nuclear stain (C and D). Proliferation was graphed as PCNA-positive cells over DAPI-positive cells (G). Sections from wild-type (E) and transgenic (F) littermates were stained by TUNEL to assess apoptosis. Bars, 100 μM.

Article Snippet: Sections were counterstained with DAPI (Vector Laboratories), and values were expressed as the number of proliferating osteoblasts over the number of DAPI-positive cells.

Techniques: Transgenic Assay, Staining, TUNEL Assay