Journal: Scientific Reports

Article Title: Phenotypic analysis of Myo10 knockout ( Myo10 tm2/tm2 ) mice lacking full-length (motorized) but not brain-specific headless myosin X

doi: 10.1038/s41598-018-37160-y

Figure Lengend Snippet: High penetrance of persistence of the hyaloid vasculature, without retinal hypovascularization, in Myo10 tm2/tm2 mice. ( A ) High-resolution MRI (magnetic resonance imaging) of enucleated and fixed eyes from adult wild-type (WT) and Myo10 tm2/tm2 mice (representative of 6 eye scans for each genotype) reveals persistence of the hyaloid vasculature in mutant mice. The hyaloid artery emerges from the optic disc and extends towards the lens, as schematically illustrated on the right. Scale bars: 1 mm. ( B ) Whole-mount retina from an adult WT and Myo10 tm2/tm2 mouse stained with anti-collagen, type IV antibodies. Hyaloid vessels can be seen emerging from the central optic disc of the Myo10 tm2/tm2 retina. Scale bar: 1 mm. ( C ) Schematic diagram showing putative roles of Myo10 in macrophage-to-endothelial cell Wnt signaling, and knockout mouse models in which hyaloid persistence phenotypes, with or without retinal hypovascularization, manifest. Notably, Wnt7b d1 is a hypomorphic allele, implying that there is reduced, but not absent, Wnt7b gene function in Wnt7b d1/d1 mice. PU.1 null (PU.1−/−) mice lack macrophages.

Article Snippet: After washing (2 × 5 min) in PBlec buffer (1 mM CaCl 2 , 1 mM MgCl 2 , 0.1 mM MnCl 2 and 0.4% Triton X-100 in PBS), each retina was incubated overnight at 4 °C with 1:25 Biotinylated GSL I-B 4 isolectin ( Griffonia Simplicifolia Lectin I isolectin (GSL I) B4; B-1205, Vector Laboratories, Burlingame, CA) and 1:200 rabbit anti-mouse collagen IV antibodies (2150-1470; Bio-Rad, Oxford, United Kingdom) diluted in PBlec buffer.

Techniques: Magnetic Resonance Imaging, Mutagenesis, Staining, Knock-Out

Journal: Scientific Reports

Article Title: Phenotypic analysis of Myo10 knockout ( Myo10 tm2/tm2 ) mice lacking full-length (motorized) but not brain-specific headless myosin X

doi: 10.1038/s41598-018-37160-y

Figure Lengend Snippet: Myo10 expression in retinal vascular endothelial cells and retinal vascularization in Myo10 tm2/tm2 mice. ( A ) Expression profile of the myosin superfamily in postnatal retinal endothelial cells. Notably, RNA sequence analysis revealed increased expression of Myo10 between postnatal day 6 (P6) and P15 (n = 3 independent preparations per group). ( B ) Schematic diagram showing vascularization of the deeper layers of the retina between P7 and P15. ( C ) Whole-mount retinas from wild-type (WT) and Myo10 tm2/tm2 mice stained for blood vessels (representative of 6 preparations for each genotype). The white arrow indicates persistent hyaloid vessels. Quantitative analysis of the vascularization of WT versus Myo10 tm2/tm2 retinas is shown on the right (n = 6 retinal preparations (from 3 mice) for each group). Vascular area is the percentage of area covered by the vascular network and lacunarity is a measure of heterogeneity. Scale bars: 1 mm. ( D ) Higher magnification images of isolectin B4-stained retinal blood vessels. Quantitative analysis of vessel branching (branch points/unit area) is shown on the right. Scale bars: 500 µm. ( E ) Higher magnification images (obtained using a 20x/0.45 objective lens) of retinal vascular layers, color coded blue (deep plexus), green (intermediate (inter.) plexus) and red (superficial (superf.) plexus). Scale bars: 50 µm. ( F ) Side-view projections of 3D reconstructions of retinal vessels, obtained by spinning disk confocal microscopy (via a 20x/0.45 objective lens). Anti-collagen, type IV, stained the superficial plexus (red), whereas all vessels were labeled with isolectin B4 (green). Scale bars: 100 µm.

Article Snippet: After washing (2 × 5 min) in PBlec buffer (1 mM CaCl 2 , 1 mM MgCl 2 , 0.1 mM MnCl 2 and 0.4% Triton X-100 in PBS), each retina was incubated overnight at 4 °C with 1:25 Biotinylated GSL I-B 4 isolectin ( Griffonia Simplicifolia Lectin I isolectin (GSL I) B4; B-1205, Vector Laboratories, Burlingame, CA) and 1:200 rabbit anti-mouse collagen IV antibodies (2150-1470; Bio-Rad, Oxford, United Kingdom) diluted in PBlec buffer.

Techniques: Expressing, Sequencing, Staining, Confocal Microscopy, Labeling