A new animal model for age-related macular degeneration
Supervisor: Paulo Pereira
Rotation Code: PP1
macular degeneration (AMD) is the leading cause of blindness in the
western world in the population over 60 years of age. Oxidative stress
and inflammation have been implicated in the pathogenesis of AMD.
However, the molecular mechanisms that associate oxidative stress to
inflammation in AMD are still unclear. In our laboratory, we have been
interested in the possibility that age-related impairment of the
proteasome activity by increased oxidative stress might be a
mechanistic link between retinal oxidative insult and expression of
AMD-like phenotypes. Indeed, our recent work indicates that impairment
of the proteasome in retinal pigment epithelial (RPE) cells results in
At present there are no animal models of AMD that reproduce the most relevant features of the disease. During this rotation you will have the oportunity to participate in the development of an animal model of disease. The rotation involves work in molecular and cell biology and you will also have the oportunity to follow some animal experimentation. The rotation is integrated in a project that aims at targeting specific proteins, such as ubiquitin, and to induce a “chronic” inactivation of the proteasome in the retina. For this purpose, a transgenic mouse expressing a mutant ubiquitin (K6W) that works as an inhibitor of proteasome-mediated proteolysis is being generated in our laboratory and you will be able to learn the basic priciples involved in generation of this animal model. A major goal of this project is to assess the effect of K6W expression in the regulation of signal transduction pathways and consequent expression of chemokines and growth factors involved in angiogenesis and inflammation in the retina. During the rotation you are expected to participate in some of the following experimental approaches: 1) To evaluate the effect of K6W expression in the activation of signaling pathways involved in inflammation and angiogenesis in the RPE of the transgenic animals by Western Blot; 2) To assess the localization and activity of transcription factors involved in angiogenesis and inflammation in the transgenic animals by immunohistochemistry and gel shift assays; 3) To determine the levels of angiogenic and inflammatory chemokines and growth factors in the transgenic animals by RT-PCR and ELISA.
Students with an interest in pre-clinical or translational reserach will have the opportunity to use some of most advanced non-invasive techniques to assess the retinal function in such animal models and will therefore be able to compare the phenotype of these animal models with the phenotype associated with the human disease.
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