Journal: eLife

Article Title: PASK links cellular energy metabolism with a mitotic self-renewal network to establish differentiation competence

doi: 10.7554/eLife.81717

Figure Lengend Snippet:

Article Snippet: Antibody , Rabbit polyclonal Anti-BrdU , Thermo Fisher Scientific , Thermo Fisher Scientific Cat# PA5-32256, RRID: AB_2549729 , IF: 1:100.

Techniques: Isolation, Software

Journal: Frontiers in Endocrinology

Article Title: Ontogeny of ependymoglial cells lining the third ventricle in mice

doi: 10.3389/fendo.2022.1073759

Figure Lengend Snippet: Characterization of BrdU-positive cells on the rostrocaudal axis. (A–C) Low- (x20, A) and high- (x63, B-C) magnification z-stack images showing the colocalization of vimentin immunoreactivity (yellow) and BrdU (pink) in zone 2 with Dapi counterstaining (white) in a coronal section from P21 male pups after BrdU injection to pregnant dams at E13. (D–F) Low- (x20, D) and high- (x63, E-F) magnification z-stack images showing the colocalization of NeuN immunoreactivity (yellow) and BrdU (pink) with Dapi counterstaining (white) in a coronal section at E13 in zone 2. (G–J) Low-magnification z-stack images (20x) showing the distribution of BrdU immunoreactivity (pink) with Dapi counterstaining (white) in coronal sections at E13 in zone 1 (G) , zone 2 (H) , zone 3 (I) , and zone 4 (J) . “E13” indicates the BrdU injection time point. Single arrowheads point out a BrdU labeling filling out the entire cell nucleus, whereas the double arrowheads point out a labeling that does not. ARH, arcuate nucleus of the hypothalamus; DMH, dorsomedial nucleus of the hypothalamus; ME, median eminence; VMH, ventromedial nucleus of the hypothalamus; 3V, third ventricle.

Article Snippet: The rabbit polyclonal antibody to BrdU (Bio-rad Cat#AHP2405, RRID: AB_2922993) recognizes the synthetic thymidine analog bromodeoxyuridine (BrdU).

Techniques: Injection, Labeling

Journal: Frontiers in Endocrinology

Article Title: Ontogeny of ependymoglial cells lining the third ventricle in mice

doi: 10.3389/fendo.2022.1073759

Figure Lengend Snippet: Characterization of BrdU-positive cells in time. (A–D) Low-magnification z-stack images (20x) showing the distribution of BrdU immunoreactivity (pink) in zone 2 (bregma -1.8) with Dapi counterstaining (white) in coronal sections from P21 male pups after BrdU injection to pregnant dams at E12 (A) , E13 (B) , E14 (C) , and E15 (D) (labeled “E12”, “E13”, “E14”, and “E15” on the pictures, respectively). The white bars point out the peaks of genesis along the third ventricle. ARH, arcuate nucleus of the hypothalamus; DMH, dorsomedial nucleus of the hypothalamus; ME, median eminence; VMH, ventromedial nucleus of the hypothalamus; 3V, third ventricle.

Article Snippet: The rabbit polyclonal antibody to BrdU (Bio-rad Cat#AHP2405, RRID: AB_2922993) recognizes the synthetic thymidine analog bromodeoxyuridine (BrdU).

Techniques: Injection, Labeling

Journal: Frontiers in Endocrinology

Article Title: Ontogeny of ependymoglial cells lining the third ventricle in mice

doi: 10.3389/fendo.2022.1073759

Figure Lengend Snippet: Developmental hypothalamic BrdU incorporation in NeuN-positive cells. (A) Stacked graph displaying the proportion of newborn BrdU/NeuN-positive cells in the hypothalamic regions vmARH, dmARH, lVMH, mVMH, cDMH, and DMH per age over all analyzed brains from P21-22 animals having received a single BrdU injection from E9 to P8 ( e.g. , “E9” brains were harvested from P21-22 male pups whose mothers received a single BrdU injection during pregnancy at E9; “E10” brains were harvested from P21-22 male pups whose mothers received a single BrdU injection during pregnancy at E10…). The bars represent the percentage of cells per age over the whole analyzed period. Within a single age, the colors represent the percentage of cells per nucleus over the entire analyzed region. (B) Number (mean ± SEM) of BrdU/NeuN-positive cells per nucleus and per age over 4 hypothalamic sections (one in each rostrocaudal zone). (C–H) Number (average -line- and individual values -dots-) of BrdU/NeuN-positive cells per rostrocaudal zones (1 to 4) and per age in each hypothalamic region. n=3 to 7 animals per group. vmARH, ventromedial arcuate nucleus of the hypothalamus; dmARH, dorsomedial arcuate nucleus of the hypothalamus; lVMH, lateral ventromedial nucleus of the hypothalamus; mVMH, medial ventromedial nucleus of the hypothalamus; cDMH, compact dorsomedial nucleus of the hypothalamus; DMH, dorsomedial nucleus of the hypothalamus. See Figure S1 .

Article Snippet: The rabbit polyclonal antibody to BrdU (Bio-rad Cat#AHP2405, RRID: AB_2922993) recognizes the synthetic thymidine analog bromodeoxyuridine (BrdU).

Techniques: BrdU Incorporation Assay, Injection

Journal: Frontiers in Endocrinology

Article Title: Ontogeny of ependymoglial cells lining the third ventricle in mice

doi: 10.3389/fendo.2022.1073759

Figure Lengend Snippet: Characterization of BrdU-positive ependymoglial cells along the ventricular wall. (A) Schematic representation of zone 3 in the mediobasal hypothalamus. Rectangles display the different ependymal regions used for the analysis: beta-1 tanycytes facing the lateral ME and the vmARH (β1 Tan), alpha-2 tanycytes facing the dmARH (α2 Tan), alpha-1 tanycytes facing the cDMH (α1 Tan), and typical ependymal cells facing the dorsal part of the DMH. (B–E) High-magnification z-stack (63x) illustrative coronal images showing the distribution of BrdU immunoreactivity (pink) in the typical ependymal cells (B) , α1 (C) , α2 (D) , and β1 (E) from the developmental timepoint E13 to E17. DAPI counterstaining is represented in white. The white bars point out the respective cells of interest along the third ventricle. ARH, arcuate nucleus of the hypothalamus; DMH, dorsomedial nucleus of the hypothalamus; ME, median eminence; V3, third ventricle. The scale bar is shown in the figure. See File S1 .

Article Snippet: The rabbit polyclonal antibody to BrdU (Bio-rad Cat#AHP2405, RRID: AB_2922993) recognizes the synthetic thymidine analog bromodeoxyuridine (BrdU).

Techniques:

Journal: Frontiers in Endocrinology

Article Title: Ontogeny of ependymoglial cells lining the third ventricle in mice

doi: 10.3389/fendo.2022.1073759

Figure Lengend Snippet: Developmental hypothalamic BrdU incorporation in vimentin (Vim)-positive cells. (A) Stacked graph displaying the proportion of newborn BrdU/Vim-positive cells in typical ependymal and tanycyte populations per age over all analyzed brains from P21-22 animals having received a single BrdU injection from E9 to P8 ( e.g. , “E9” brains were harvested from P21-22 male pups whose mothers received a single BrdU injection during pregnancy at E9; “E10” brains were harvested from P21-22 male pups whose mothers received a single BrdU injection during pregnancy at E10…). The bars represent the percentage of cells per age over the whole analyzed period. Within a single age, the colors represent the percentage of cells per ependymal subpopulation over the whole analyzed region. (B) Number (mean ± SEM) of BrdU/Vim-positive cells per ependymal subpopulations and per age over 4 hypothalamic sections (one in each rostrocaudal zone). (C–H) Number (average -line- and individual values -dots-) of BrdU/Vim-positive cells per rostrocaudal zones (1 to 4) and per age in each ependymal subpopulation. n=3 to 7 animals per group. α1 Tan, alpha-1 tanycytes; α2 Tan, alpha-2 tanycytes; β1v Tan, ventral beta-1 tanycytes; β1d Tan, dorsal beta-1 tanycytes; β2 Tan, beta-2 tanycytes. See Figure S2 .

Article Snippet: The rabbit polyclonal antibody to BrdU (Bio-rad Cat#AHP2405, RRID: AB_2922993) recognizes the synthetic thymidine analog bromodeoxyuridine (BrdU).

Techniques: BrdU Incorporation Assay, Injection

Journal: Frontiers in Endocrinology

Article Title: Ontogeny of ependymoglial cells lining the third ventricle in mice

doi: 10.3389/fendo.2022.1073759

Figure Lengend Snippet: Cellular and molecular hypothalamic generation of mature tanycytes. (A) Representative plot illustrating the patterns of neuron, ependymal, and tanycyte generation across embryonic and early postnatal development extrapolated from our quantifications. (B) Rostrocaudal neuroanatomical representations of the developmental peaks of BrdU integration along the ventricular wall and hypothalamic nuclei. The ages indicate the peaks of genesis. The symbol ø indicates no clear peak of genesis ( i.e ., either multiple peaks or a continuous generation). (C) UMAP plots showing neuron (pink), ependymal (blue), and tanycyte-like (green) clusters in diencephalic and hypothalamic scRNAseq datasets , from E13 to P45, per developmental time point. (D) Violin plot showing specific features in tanycyte-like clusters across developmental time points from E14 to P45. (E) Comparison between the expression of the top 100 features of tanycyte-like cells at P45 versus those of tanycyte-like cells from every other developmental time point. The black dots represent features regarding their -log10(pAdj) significance. The blue line displays the number of features in the top 100 at each timepoint shared with the top 100 at P45. (F) Plot showing the comparison between the 100 more enriched GO terms from tanycyte-like cells at P45 (based on FDR) versus GO terms found in tanycyte-like cells from every other developmental time point. The black dots represent GO terms regarding their -log(FDR) significance. The blue line represents the number of GO terms shared in tanycyte-like clusters compared to P45. (G) Plot showing the -log10(FDR) of the top 40 more enriched GO terms in tanycyte-like cells at P45 compared to all time points. See Figure S3 .

Article Snippet: The rabbit polyclonal antibody to BrdU (Bio-rad Cat#AHP2405, RRID: AB_2922993) recognizes the synthetic thymidine analog bromodeoxyuridine (BrdU).

Techniques: Comparison, Expressing