15 research outputs found

    Retinal Expression of Wnt-Pathway Mediated Genes in Low-Density Lipoprotein Receptor-Related Protein 5 (Lrp5) Knockout Mice

    Get PDF
    Mutations in low-density lipoprotein receptor-related protein 5 (Lrp5) impair retinal angiogenesis in patients with familial exudative vitreoretinopathy (FEVR), a rare type of blinding vascular eye disease. The defective retinal vasculature phenotype in human FEVR patients is recapitulated in Lrp5 knockout (Lrp5βˆ’/βˆ’)(Lrp5^{-/-}) mouse with delayed and incomplete development of retinal vessels. In this study we examined gene expression changes in the developing Lrp5βˆ’/βˆ’Lrp5^{βˆ’/βˆ’} mouse retina to gain insight into the molecular mechanisms that underlie the pathology of FEVR in humans. Gene expression levels were assessed with an Illumina microarray on total RNA from Lrp5βˆ’/βˆ’Lrp5^{βˆ’/βˆ’} and WT retinas isolated on postnatal day (P) 8. Regulated genes were confirmed using RT-qPCR analysis. Consistent with a role in vascular development, we identified expression changes in genes involved in cell-cell adhesion, blood vessel morphogenesis and membrane transport in Lrp5βˆ’/βˆ’Lrp5^{βˆ’/βˆ’} retina compared to WT retina. In particular, tight junction protein claudin5 and amino acid transporter slc38a5 are both highly down-regulated in Lrp5βˆ’/βˆ’Lrp5^{βˆ’/βˆ’} retina. Similarly, several Wnt ligands including Wnt7b show decreased expression levels. Plasmalemma vesicle associated protein (plvap), an endothelial permeability marker, in contrast, is up-regulated consistent with increased permeability in Lrp5βˆ’/βˆ’Lrp5^{βˆ’/βˆ’} retinas. Together these data suggest that Lrp5 regulates multiple groups of genes that influence retinal angiogenesis and may contribute to the pathogenesis of FEVR

    Resveratrol Inhibits Pathologic Retinal Neovascularization in Vldlrβˆ’/βˆ’ Mice

    No full text
    Macular telangiectasia (MacTel) is a vision-threatening retinal disease with unknown pathogenesis and no approved treatment. Very low-density lipoprotein receptor mutant mice (Vldlrβˆ’/βˆ’) exhibit critical features of MacTel, such as retinal neovascularization and photoreceptor degeneration. In this study, the authors evaluated the therapeutic potential of resveratrol, a plant polyphenol, in Vldlrβˆ’/βˆ’ mice as a model for MacTel

    Selected groups of genes regulated in <i>Lrp5<sup>βˆ’/βˆ’</sup></i> retina.

    No full text
    <p>Note: Retinas were isolated from P8 <i>Lrp5<sup>βˆ’/βˆ’</sup></i> mice and age matched WT control mice. RNA was isolated and assessed with Illumina gene expression microarray. (βˆ’) indicates decreased expression in <i>Lrp5<sup>βˆ’/βˆ’</sup></i> retina and (+) indicates increased expression in <i>Lrp5<sup>βˆ’/βˆ’</sup></i> retina compared to WT control.</p

    Regulation of tight junction, membrane transport, angiogenic, and cell adhesion genes in the <i>Lrp5</i> null retina.

    No full text
    <p>Heat maps illustrate the results of a gene array run from whole retinal total mRNA. The most regulated families of genes were (A) claudin family genes, (B) membrane transport genes, (C) angiogenic regulatory genes, and (D) cell adhesion/cell-cell junction genes. Each sample is represented by a block: either wild-type (WT) samples 1 through 3, and Lrp5 null samples 1–3. Relative down-regulation of expression in <i>Lrp5</i> null retina compared to WT retina is represented by green, while relative up-regulation is in red. No relative regulation is black (See scale on Figure).</p

    Common genes regulated in <i>Lrp5<sup>βˆ’/βˆ’</sup></i> and <i>Norrin<sup>βˆ’/βˆ’</sup></i> retinas.

    No full text
    <p>Note: Retinas were isolated from P8 <i>Lrp5<sup>βˆ’/βˆ’</sup></i> mice and age matched WT control mice. RNA was isolated and assessed with Illumina gene expression microarray. (βˆ’) indicates decreased expression in <i>Lrp5<sup>βˆ’/βˆ’</sup></i> retina compared to WT retina, and (+) indicates increased expression in <i>Lrp5<sup>βˆ’/βˆ’</sup></i> retina.</p><p>*: Genes regulated in P7 <i>Norrin<sup>βˆ’/βˆ’</sup></i> retina were adapted from Schafer et. al. IOVS, 2009 <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030203#pone.0030203-Schafer1" target="_blank">[33]</a>.</p

    Abnormal vascular development in the inner and outer retina and forebrain of <i>Lrp5</i> null mice.

    No full text
    <p>(A) Retinal whole mounts of WT and <i>Lrp5</i> null mice stained for endothelial cells with isolectin B<sub>4</sub>-594 at P12 and 17. Enlarged images highlight the abnormal vessel growth in the <i>Lrp5</i> null retina. (B) Retinal cross sections from P30 WT and <i>Lrp5</i> null mice stained for endothelial cells with isolectin B<sub>4</sub>-594 (red) and cell nuclei with DAPI (blue). Arrows indicate deep layers of retinal vasculature which is present in WT retina but absent in <i>Lrp5</i> null retina. (C) Cross sections of the forebrain from P8 WT and <i>Lrp5</i> null mice with endothelial cells stained with isolectin B<sub>4</sub>-594. Scale bar: 100 Β΅m.</p

    Delayed development of the superficial retinal vasculature and persistent hyaloid vessels in <i>Lrp5</i> null mice.

    No full text
    <p>(A) Left: retinal whole mounts stained with isolectin B<sub>4</sub>-594 from WT and <i>Lrp5</i> null mice at post-natal day (P) 8. Right: quantification of vascularized retinal area in WT and <i>Lrp5</i> null mice at P8. nβ€Š=β€Š5–10 per group, ***<i>p</i><0.001. (B) Retinal cross sections of WT mice and <i>Lrp5</i> null mice at P8 stained for endothelial cells with isolectin B<sub>4</sub>-594 (red) and cell nuclei (DAPI, blue). Arrows indicate persistent hyaloid vessels in <i>Lrp5</i> null retina. Scale bars: 500 Β΅m.</p

    Expression levels of <i>Lrp5</i>, <i>Norrin</i>, <i>Frizzled4</i> and <i>Dvl</i> mRNA during retinal development in wild type mice.

    No full text
    <p>Quantification of mRNA (A) <i>Lrp5</i>, (B) <i>Norrin</i>, (C) <i>Frizzled4</i>, (D) <i>Dvl1</i>, (E) <i>Dvl2</i>, and (F) <i>Dlv3</i> was performed with RT-qPCR during normal retinal development from P1-P17. Expression levels were normalized against house keeping gene <i>Cyclophilin A</i>. Trend lines were fitted with polynomial, linear or power regression.</p
    corecore