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Master's Dissertation
DOI
https://doi.org/10.11606/D.42.2019.tde-19022020-132653
Document
Author
Full name
Aline Martins de Almeida
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2019
Supervisor
Committee
Debbio, Carolina Beltrame Del (President)
Coltri, Patricia Pereira
Silva, Graziela Batista da
Wu, Davi Chen
Title in Portuguese
Caracterização das microvesículas nas células do epitélio ciliar de roedores.
Keywords in Portuguese
Degeneração Retiniana
Epitélio Ciliar
Vesículas Extracelulares
Abstract in Portuguese
O aumento da expectativa de vida dos seres humanos favoreceu o aumento do diagnóstico de doenças crônico-degenerativas, dentre elas as que acometem o sistema visual. Uma vez que as células neuronais não são capazes de regeneração espontânea, diversas patologias como a Retinose Pigmentar (RP), glaucoma, Degeneração Macular Relacionada com a Idade (DMRI), dentre outras, podem levar a lesão tecidual e cegueira. O Epitélio Ciliar (EC), um tecido adjacente à retina, tem se demonstrado uma fonte promissora de células-tronco/progenitoras retinianas, uma vez que estas células podem ser reprogramadas em células-tronco e se diferenciarem em neurônios retinianos in vitro. Apesar desse potencial, estas células apresentam capacidade regenerativa limitada, que pode ser controlada por fatores e sinalizações contidas no microambiente. As vesículas extracelulares (VE) são sintetizadas e liberadas por células ao ambiente e podem conter componentes sinalizadores, inclusive inibitórios da capacidade de reprogramação. Este trabalho teve como objetivo estudar a maquinaria de síntese de VEs presentes nas células do EC, assim como caracterizar as VEs liberadas por estas células quanto a sua quantidade, tamanho e morfologia. Além disso, também investigamos a capacidade de liberação de VEs em modelo animal de degeneração retiniana (p23H) em comparação com animais sem degeneração. Nossos resultados indicaram que as células do EC apresentaram microvesículas em seu citoplasma e que expressaram os transcritos dos marcadores mais comuns da biogênese de microvesículas (Alix, CD63, CD81, Anexina V, EpCAM, ICAM, TSG101 e HSP70). A análise de Rastreamento de Nanopartículas (tamanho e concentração) indicaram que as células do EC foram capazes de liberar VEs de diferentes tamanhos, variando entre 100 e 1000 nm. Os animais modelos de degeneração retiniana expressaram menos marcadores de VEs em comparação com os animais controles, porém, as vesículas liberadas apresentaram maior tamanho. Nossos resultados sugerem que as VEs das células do EC podem ser um mecanismo de comunicação celular e a degeneração retiniana podem regular a síntese e liberação destas VEs.
Title in English
Characterization of microvesicles in rodent ciliary epithelial cells.
Keywords in English
Ciliary Epithelium
Extracellular vesicles
Retinal Degeneration
Abstract in English
The increased life expectancy of humans favored the increased diagnosis of chronic degenerative diseases, including those affecting the visual system. Since neuronal cells are not capable of spontaneous regeneration, different pathologies such Retinitis Pigmentosa (RP), glaucoma, Age Macular and Degeneration (AMD), can lead to tissue damage and blindness. Ciliary Epithelium (CE), an epithelial tissue adjacent to the retina, is a promising source of retinal stem/progenitor cells, as these cells can be reprogrammed into stem cells and differentiate into retinal neurons in vitro,. Despite this potential, these cells have limited regenerative capacity, which can be controlled by factors and signals in the microenvironment. Extracellular vesicles (EVs) are synthesized and released by cells into the environment and may contain signaling components, including inhibitors of reprogramming ability. This work aimed to study the synthesis machinery of EVs present in the cells of the EC, as well as characterize its releases by these cells in terms of quantity, size and morphology. In addition, we also investigated EVs release capacity in an animal model of retinal degeneration (p23H) compared to animals without degeneration. Our results indicated that CE cells presented microvesicles in their cytoplasm and expressed the transcripts of the most common microvesicle biogenesis markers (Alix, CD63, CD81, Annexin V, EpCAM, ICAM, TSG101 and HSP70). Nanoparticle Tracking Analysis (size and concentration) indicated that CE cells were able to release EVs of different sizes, ranging from 100 to 1000 nm. Retinal degeneration model animals expressed fewer EVs markers compared to control animals, but released vesicles were larger. Our results suggest that EVs from EC cells may be a mechanism of cellular communication, and retinal degeneration may regulate the synthesis and release of these EVs.
 
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Release Date
2023-05-06
Publishing Date
2021-10-13
 
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