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Characterization of lysossome dysfunction in age-related macular degeneration
Portugal ( PorMSIC) - Nova University of Lisbon, Lisbon
CEDOC- Centro de Estudos de Doenças Crónicas Rua Câmara Pestana nº 6, 6-A Edifício CEDOC II 1150-082 Lisboa
Prof. Miguel Seabra
Type of Research Project
- Basic science
What is the background of the project?
Declining lysosomal function is postulated to cause and/or facilitate a number of chronic age-related disorders. We hypothesise that dysfunction of the lysosomal network is a critical component in the pathogenesis of age-related macular degeneration (AMD), the most common blinding disease in the western world. We developed methods to induce lysosomal dysfunction in retinal pigment epithelium (RPE) cell culture systems. We are using these induced cellular systems to study how lysosomal dysfunction reflects on other organelles and cellular functions such as mitochondria, lysosome, autophagy, and proteostasis.
What is the aim of the project?
This project aims to dissect lysosomal dysfunction role on AMD and to develop strategies to restore lysosomal function.
What techniques and methods are used?
The project will make use of the following techniques: i) Isolation, growth and maintenance of primary cultures of porcine RPE (Retinal Pigment Epithelial cells); ii) Cell viability assays by Flow cytometry and enzymatic assays; iii) Fluorescence imaging using light and confocal microscopy; iv) Protein levels evaluation by SDS-page (sodium dodecyl sulfate-polyacrylamine gel electrophoresis) and Western blot v) Flow cytometry analysis; vi) Data analysis with sppecific software such as ImageJ, Zen and Image Lab.
What is the role of the student?
What are the tasks expected to be accomplished by the student?
The student is expected to observe and collaborate actively during the internship. He/she will be able to collaborate with the rest of the team on the isolation and growth of primary cultures of porcine retinal pigment epithelium (RPE), fluorescence imaging using light and confocal microscopy and western blot. The student will also be involved in the data analysis using some specific software such as ImageJ, Zen and Image Lab.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
The student will be able to attend lab meetings and should read the articles mentioned in this form.
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- The student will prepare a presentation - The student will prepare a scientific report - The student will prepare an abstract
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts: - Medical students - Graduated students (less than 6 months) - Pre-Medical students from the American-British system
- Keeling et al.; Cells. 2018 Feb 23;7(2). pii: E16. doi: 10.3390/cells7020016.
- Guha et al.; Exp Eye Res. 2014 Sep;126:68-76. doi: 10.1016/j.exer.2014.05.013
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