Journal: The Journal of comparative neurology

Article Title: Expression profile analysis of vulnerable CA1 pyramidal neurons in young-middle aged Ts65Dn mice

doi: 10.1002/cne.23663

Figure Lengend Snippet: (A) BDNF showed no difference in mRNA expression levels, however, the BDNF receptor TrkB (NTRK2) showed significant downregulation, in accordance with the microarray results. NTF3 showed a trend for downregulation and the NTF3 receptor TrkC (NTRK3) was significantly downregulated. ddCT was expressed as percent of 2N control for all qPCR results. Key, (* p < 0.05) and (t p < 0.1). Error bars indicate SEM.

Article Snippet: Taqman qPCR primers (Life Technologies) were utilized for qPCR, TrkB (Mm01341760_m1; targeting the exon 16-17 boundary), TrkC (Mm01317842_m1; targeting the exon 12-13 boundary), NTF3 (Mm00435413_s1; targeting exon 3), BDNF (Mm04230607_s1; targeting exon 8) and the housekeeping gene succinate dehydrogenase complex, subunit A, flavoprotein (SDHA; Mm01352360_m1; targeting the exon 14-15 boundary).

Techniques: Expressing, Microarray

Journal: Angiogenesis

Article Title: Adgrf5 contributes to patterning of the endothelial deep layer in retina

doi: 10.1007/s10456-019-09674-0

Figure Lengend Snippet: Adgrf5 is exclusively endothelial in postnatal retina. a mRNA expression levels of Adgrf5 (black line) and Pecam -1 (red line) in mouse retina. The whole retinal RNA from C57BL6 mice was used to determine Adgrf5 and Pecam - 1 transcripts by qRT-PCR at P1 ( n = 6 mice), P4 ( n = 4), P7 ( n = 3), P14 ( n = 5), P21 ( n = 5), 1 month ( n = 3), and 3 months ( n = 4). The expression of each gene was normalized by Hprt . Error bars represent ± SD. b Relative mRNA expression level of Adgrf5 normalized by Pecam - 1 in C57/Bl6 retinae. The same number of animals mentioned in Fig. a was used. c Adgrf5 mRNA expression level over the course of the eye development. Adgrf5 WT and Adgrf5 ECKO mouse retinal RNA was prepared at the time indicated and Adgrf5 transcript was assessed by qRT-PCR ( Adgrf5 WT ( n = 6) and Adgrf5 ECKO ( n = 3) at P7 and Adgrf5 WT ( n = 10) and Adgrf5 ECKO ( n = 4) at P21). Results are normalized by Hprt expression. d Fluorescent images of ADGRF5 expression in-between an artery and a vein in superficial (upper) and inner (lower) vascular layer. Gpr116 -mCherry reporter mouse retinae were collected at the time indicated, and stained with IB4 (gray) to visualize the endothelium. mCherry indicates Adgrf5 expression (magenta). e Quantification of the fluorescence intensity in Gpr116 -mCherry reporter mouse retinae, normalized over isolectin (lB4) fluorescence

Article Snippet: The following probes were used: mouse Adgrf5 (Assay: Mm01269028_m1, FAM, spanning exon 17–18, Life Technologies), Pecam -1 (Assay: Mm01242576_m1, FAM, Life Technologies), Hprt (Assay: Mm03024075_m1, FAM, Life Technologies).

Techniques: Expressing, Quantitative RT-PCR, Staining, Fluorescence

Journal: Angiogenesis

Article Title: Adgrf5 contributes to patterning of the endothelial deep layer in retina

doi: 10.1007/s10456-019-09674-0

Figure Lengend Snippet: Adgrf5 deficiency leads to perivenous abnormalities. a Involvement of Adgrf5 in the inner plexus formation. Superficial (left) and inner (middle left) vascular plexus in-between an artery and a vein from WT (upper) and Adgrf5 KO (lower) retina at P9 are shown. The endothelium is visualized by Cldn5 -GFP reporter (green) and IB4 staining (gray). Complete 3D reconstruction of the same area is represented in depth coding with a top view (middle), and a vertical view below the vein (V) and an artery (A) is indicated (right panel). b Quantification of the number of extensions from the superficial to the inner vascular layer in WT and Adgrf5 KO under the arteries (top) and under the veins (bottom). c Role of EC-specific Adgrf5 in the inner plexus development. Superficial (left) and inner (middle left) vascular layer in-between an artery and a vein in the WT (upper) and Adgrf5 ECKO (lower) retina at P10 are shown. The endothelium is visualized by IB4 staining (gray). Adgrf5 -depleted EC are marked by GFP (green). Complete 3D reconstruction of the area from WT and Adgrf5 ECKO is represented in depth coding with a top view (middle), and a vertical view below the vein (V) and an artery (A) is indicated (right panel). d Quantification of the EC protrusions from the superficial to the inner vascular layer in WT and Adgrf5 ECKO under the arteries (top) and under the veins (bottom). e Role of EC-specific Adgrf5 in the inner plexus development. Superficial (left) and inner (middle left) vascular layer in-between an artery and a vein in the WT (upper) and Adgrf5 ECGOF (lower) retina at P9 are shown. The endothelium is visualized by IB4 staining (gray). Complete 3D reconstruction of the area from WT and Adgrf5 ECGOF is represented in depth coding with a top view (middle), and a vertical view below the vein (V) and an artery (A) is indicated (right panel). f Quantification of the EC protrusions from the superficial to the inner vascular layer in WT and Adgrf5 ECGOF under the arteries (top) and under the veins (bottom)

Article Snippet: The following probes were used: mouse Adgrf5 (Assay: Mm01269028_m1, FAM, spanning exon 17–18, Life Technologies), Pecam -1 (Assay: Mm01242576_m1, FAM, Life Technologies), Hprt (Assay: Mm03024075_m1, FAM, Life Technologies).

Techniques: Staining

Journal: Angiogenesis

Article Title: Adgrf5 contributes to patterning of the endothelial deep layer in retina

doi: 10.1007/s10456-019-09674-0

Figure Lengend Snippet: Adgrf5 KO retina develops transient neovessels in the inner retinal space. a Inner vascular layer in a leaf of WT mouse retina at P12, P14, P21, 1 month, and 4 months. The endothelium is visualized in green tracking Claudin5 -GFP. Horizontal view (upper panel) and optical cross-section (lower panel) are presented. Asterisks indicate areas of high vascular densities. b Inner vascular layer in a leaf of Adgrf5 KO mouse retina at P12, P14, P21, 1 month, and 4 months. The endothelium is visualized in green tracking Claudin5 -GFP. Horizontal view (upper panel) and optical cross-section (lower panel) are presented. c Images from Adgrf5 WT (top row) and Adgrf5 KO (lower row) mouse retina at P8, P14, P21, and 1 month. Cross-sectioned retinae were stained with collagen IV (red) and Hoechst (blue). The vasculature is depicted by IB4 staining (gray). d Quantification of the vascular densities in the inner plexus from Adgrf5 WT or KO mouse retina at P12, P14, P21, and 1 month. e Quantification of the capillary perimeters in the inner plexus from Adgrf5 WT or KO mouse retina at P14. f Quantification of the subretinal neovascular sprouts emerging from the inner plexus in Adgrf5 WT or KO whole mouse retina at P14. g 3D reconstruction images of the three vascular layers below the vein of Adgrf5 KO retina at P14. The retinal structure is presented in depth coding, displaying EC extensions after the inner vascular layer in the subretinal space (deep sprout is marked by an arrowhead). h Superficial (left) and inner (middle left) retinal vascular layer in-between an artery and a vein in WT and Adgrf5 KO (normal and severe) mice at P14. The 3D reconstruction images of the three vascular layers and the neovessels below (middle right) are shown. IB4 (gray) and Cldn5 -GFP (green) were used to label the endothelium, followed by ASMA staining to distinguish the artery and the vein. Vertical 3D projection of the three vascular layers and the neovessels below in the Adgrf5 KO at high magnification (×63) (right) (deep sprout is marked by an arrowhead). i Superficial (left), inner (middle) vascular plexus and 3D view (right) from WT and Adgrf5 Δ17 KO at P14. The vascular layers were stained with IB4 (gray) (deep sprout is marked by an arrowhead). j Superficial (left), inner (middle) vascular plexus and 3D view (right) from WT and Adgrf5 ECKO at P14. The vascular layers were stained with IB4 (gray) (deep sprout is marked by an arrowhead). k Superficial (left), inner (middle) vascular plexus and 3D view (right) from WT and Adgrf5 ECGOF at P14. The vascular layers were stained with IB4 (gray) (deep sprout is marked by an arrowhead)

Article Snippet: The following probes were used: mouse Adgrf5 (Assay: Mm01269028_m1, FAM, spanning exon 17–18, Life Technologies), Pecam -1 (Assay: Mm01242576_m1, FAM, Life Technologies), Hprt (Assay: Mm03024075_m1, FAM, Life Technologies).

Techniques: Staining

Journal: Angiogenesis

Article Title: Adgrf5 contributes to patterning of the endothelial deep layer in retina

doi: 10.1007/s10456-019-09674-0

Figure Lengend Snippet: Adgrf5 KO retinal tissue recovers after vessel normalization. a Images from WT and Adgrf5 KO mouse retina at P14. Cross-sectioned retinae were stained with H&E (top) and Hoechst (blue). The vasculature is depicted by IB4 staining (gray) and Claudin5 -GFP expression (green). b 3D reconstruction images of the three vascular layers of Adgrf5 KO retina at P14. The retinal structure is presented viewed from the superficial layer (3D top view, left), or from a vertical view (right). Erythrocytes are visualized by Ter119 staining. c Images from WT and Adgrf5 KO mouse retina at P8, P14, P21, and 1 month. Cross-sectioned retinae were stained with anti-NeuN antibody (green), Hoechst (blue), and IB4 (gray). d Images from WT and Adgrf5 KO mouse retina at P8, P14, P21, and 1 month. Cross-sectioned retinae were stained with anti-Glutamine synthetase antibody (green), Hoechst (blue), and IB4 (gray). e Images from WT and Adgrf5 KO mouse retina at P8, P14, P21, and 1 month. Cross-sectioned retinae were stained with anti-GFAP antibody (red), Hoechst (blue), and IB4 (gray)

Article Snippet: The following probes were used: mouse Adgrf5 (Assay: Mm01269028_m1, FAM, spanning exon 17–18, Life Technologies), Pecam -1 (Assay: Mm01242576_m1, FAM, Life Technologies), Hprt (Assay: Mm03024075_m1, FAM, Life Technologies).

Techniques: Staining, Expressing

Journal: Current biology : CB

Article Title: Quantitative Control of GPCR Organization and Signaling by Endocytosis in Epithelial Morphogenesis

doi: 10.1016/j.cub.2018.03.068

Figure Lengend Snippet: (A) Apical projections of ectoderm tissue expressing E-cad::GFP (green) and MyoIIRLC::mCherry (magenta) at 10 min post-cellularization in control, Gprk2-shRNA, and β-arrestin-2-shRNA. Scale bar, 5 μm.

Article Snippet: RT-qPCR, Gprk2: Dm02134696_g1 (exon 2-3 boundary, 1192, 101bp) , This paper, Lecuit Laboratory , N/A.

Techniques: Expressing, Control, shRNA

Journal: Current biology : CB

Article Title: Quantitative Control of GPCR Organization and Signaling by Endocytosis in Epithelial Morphogenesis

doi: 10.1016/j.cub.2018.03.068

Figure Lengend Snippet: (A) Distribution of Rho1:GTP sensor or Anilin Rho-binding domain fused with GFP (AniRBD::GFP) (green) 10 min into germband extension in control, Gprk2-shRNA, and β-arrestin-2-shRNA embryos. Scale bars, 5 μm.

Article Snippet: RT-qPCR, Gprk2: Dm02134696_g1 (exon 2-3 boundary, 1192, 101bp) , This paper, Lecuit Laboratory , N/A.

Techniques: Binding Assay, Control, shRNA

Journal: Current biology : CB

Article Title: Quantitative Control of GPCR Organization and Signaling by Endocytosis in Epithelial Morphogenesis

doi: 10.1016/j.cub.2018.03.068

Figure Lengend Snippet: (A–C) Images represent apical region in the cells from the ventrolateral region of the embryos. Smog::GFP embryos are injected with Dextran-568 to show colocalization in (A) control, (B) Gprk2-shRNA, and (C) β-arrestin-2-shRNA. Scale bars (A–C), 5 μm. Arrowheads (red) mark Smog localization in the Dextran-filled structures.

Article Snippet: RT-qPCR, Gprk2: Dm02134696_g1 (exon 2-3 boundary, 1192, 101bp) , This paper, Lecuit Laboratory , N/A.

Techniques: Injection, Control, shRNA

Journal: Current biology : CB

Article Title: Quantitative Control of GPCR Organization and Signaling by Endocytosis in Epithelial Morphogenesis

doi: 10.1016/j.cub.2018.03.068

Figure Lengend Snippet: (A–F) Fog levels are reduced using fog-dsRNA compared to water injections. Fog concentration is increased using overexpression with UAS-fog12 and was compared to the appropriate controls. (A–C) density/concentration and (D–F) brightness of Smog::GFP particles are calculated in (A and D) controls, (B and E) Gprk2-KD, and (C and F) β-arrestin-2-KD embryos with change in the Fog concentration, fog-dsRNA (left) and UAS-fog12 (right).

Article Snippet: RT-qPCR, Gprk2: Dm02134696_g1 (exon 2-3 boundary, 1192, 101bp) , This paper, Lecuit Laboratory , N/A.

Techniques: Concentration Assay, Over Expression

Journal: Current biology : CB

Article Title: Quantitative Control of GPCR Organization and Signaling by Endocytosis in Epithelial Morphogenesis

doi: 10.1016/j.cub.2018.03.068

Figure Lengend Snippet: (A) Trace of cell showing plasma membrane invaginations in different Z sections, with co-localization between AniRBD::GFP and Dextran-647 (marked with red arrowhead) in control, Gprk2-shRNA, and β-arrestin-2-shRNA embryos. Scale bars, 1 μm.

Article Snippet: RT-qPCR, Gprk2: Dm02134696_g1 (exon 2-3 boundary, 1192, 101bp) , This paper, Lecuit Laboratory , N/A.

Techniques: Clinical Proteomics, Membrane, Control, shRNA

Journal: Current biology : CB

Article Title: Quantitative Control of GPCR Organization and Signaling by Endocytosis in Epithelial Morphogenesis

doi: 10.1016/j.cub.2018.03.068

Figure Lengend Snippet: Key Resources Table

Article Snippet: RT-qPCR, Gprk2: Dm02134696_g1 (exon 2-3 boundary, 1192, 101bp) , This paper, Lecuit Laboratory , N/A.

Techniques: Recombinant, Electron Microscopy, Reverse Transcription, Plasmid Preparation, Software, Microscopy