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Exosomes as regulators of Proteostasis in Age-Related Macular Degeneration
Portugal ( PorMSIC) - Nova University of Lisbon, Lisbon
CEDOC- CHRONIC DISEASES RESEARCH CENTER Rua Camara Pestana no 6, 6-A, Edificio CEDOC II, 1150-082 Lisboa, Portugal
Paulo Pereira, Jo
Type of Research Project
- Basic science
What is the background of the project?
Proteostasis deregulation is one of the main molecular mechanisms involved in cellular dysfunction upon ageing. The regulation of proteostasis involves protein quality control, through the concerted action of molecular chaperones, and the lysosomal and proteasomal proteolytic systems. Nonetheless, over the years, a number of compelling evidences now suggests that exosomes may have an important role in the regulation of protesotasis. Exosomes are vesicles (30–120 nm) secreted by most cell types that can signal to distant cells, tissues or organs, by carrying and delivering RNA and proteins, thus modulating many biological functions. In this context, it has been hypothesized that the loading of exosomes with damaged and/or oligomerized proteins may constitute a mechanism for the disposal of unwanted and toxic protein material from the cells. During age-related macular degeneration, vision loss is highly associated with damaged to a monolayer of postmitotic cells that support photoreceptors called retinal pigmented epithelium (RPE). We will use these cells to study the age-related cell damaged role in the protein loading and release of exosomes.
What is the aim of the project?
This project is design to test the hypothesis that oxidative damage during aging contibutes to the disposal of toxic proteins through exosome release in the retinal pigmented epithilium.
What techniques and methods are used?
We will use cell culture techniques to produce cell and exosomal samples. Exosomes will be isolate from cell culture media of control or challenged cells. We will subsequently use western blotting and immunocytochemistry techniques to assess the levels of released exosomes and their contents.
What is the role of the student?
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
All work will be done with supervision in order to give the student enough time to understand the procedures and learn from observation. But after learning the techniques, hopefully the student will know how to perform some experiments on his/her own. If so, the student will be able to perform various different techniques, such as cell culture techniques, western blotting and immunocytochemistry techniques. The student is also expected to critically discuss the results obtained.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
Yes. The student will have access to literature regarding the state of the art of the scientific field.
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- No specific outcome is expected
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
- Fernandes AF; Zhou J; Zhang X; Bian Q; Sparrow J; Taylor A; Pereira P; Shang F. J Oxidative inactivation of the proteasome in retinal pigment epithelial cells. A potential link between oxidative stress and up-regulation of interleukin-8. Biol Chem. 2008 Jul 25;283(30):20745-53.
- Catarino S; Bento CF; Brito A; Murteira E; Fernandes AF; Pereira P. Regulation of the expression of interleukin-8 induced by 25-hydroxycholesterol in retinal pigment epithelium cells. Acta Ophthalmol. 2012 Jun;90(4):e255-63. doi: 10.1111/j.1755- 3768.2011.02350.x. Epub 2012 Feb 7.
- Soares AR; Martins-Marques T; Ribeiro-Rodrigues T; Ferreira JV Catarino S; Pinho MJ; Zuzarte M; Anjo SI; Manadas B; Sluijter JPG; Pereira P; Girao H. Gap junctional protein Cx43 is involved in the communication between extracellular vesicles and mammalian cells. Sci Reports 2015; 5:13243.
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