Publication: Autofagia selectiva en la retina: fisiología y patología
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2019-03-19
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Universidad Complutense de Madrid
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Abstract
En esta tesis doctoral hemos investigado el papel de la autofagia selectiva durante el desarrollo y degeneración del ojo de vertebrados. En primer lugar, nos hemos centrado en la mitofagia o la eliminación selectiva de mitocondrias por autofagia. Para este estudio hemos tenido primero que optimizar una serie de herramientas y métodos que nos permiten determinar de una manera cuantitativa, robusta, y lo más importante, in vivo, este proceso. Entre estos métodos destacamos el uso de dos animales reporteros para determinar autofagia y mitofagia en tejido. También se han optimizado diferentes inductores e inhibidores de autofagia y mitofagia, controlando sus cinéticas que nos permiten modular el proceso. Nuestros datos indican que la autofagia predomina en las diferentes partes del ojo durante su desarrollo. En el ojo de adulto, sin embargo, encontramos en la capa nuclear externa (CNE), donde se encuentran los fotorreceptores y la glía de Müller, el epitelio pigmentario y nervio óptico, que toda la actividad autofágica corresponde a mitofagia, mientras que en las otras partes del ojo hay autofagia pero no mitofagia. Por otro lado, hemos demostrado que para que tenga lugar la mitofagia en la CNE, hay un transporte desde el segmento interno a la zona de los somas tanto de los lisosomas como de los mitolisosomas...
In this doctoral thesis we have investigated the role of selective autophagy during the development and degeneration of the vertebrate eye. First, we have focused on the mitophagy or the selective elimination of mitochondria by autophagy. For this study, we first had to optimize a series of tools and methods that allow us to determine this process in a quantitative and robust form, and most important way, in vivo. Among these methods we highlight the use of two reporter animals to determine autophagy and mitophagy in tissue. Different inductors and inhibitors of autophagy and mitophagy have also been optimized, controlling their kinetics that allow us to modulate the process.Our data indicate that autophagy predominates in different parts of the eye during its development. In the adult eye, however, we find in the outer nuclear layer (ONL), where the photoreceptors and Müller glia, the pigmentary epithelium and the optic nerve are located, that all the autophagic activity is mitophagy, while in the other parts of the eye there is autophagy but no mitophagy. On the other hand, we have defined that to take place the mitophagy in the ONL, there is a transport from the internal segment to the area of the somas of both lysosomes and mitolysosomes...
In this doctoral thesis we have investigated the role of selective autophagy during the development and degeneration of the vertebrate eye. First, we have focused on the mitophagy or the selective elimination of mitochondria by autophagy. For this study, we first had to optimize a series of tools and methods that allow us to determine this process in a quantitative and robust form, and most important way, in vivo. Among these methods we highlight the use of two reporter animals to determine autophagy and mitophagy in tissue. Different inductors and inhibitors of autophagy and mitophagy have also been optimized, controlling their kinetics that allow us to modulate the process.Our data indicate that autophagy predominates in different parts of the eye during its development. In the adult eye, however, we find in the outer nuclear layer (ONL), where the photoreceptors and Müller glia, the pigmentary epithelium and the optic nerve are located, that all the autophagic activity is mitophagy, while in the other parts of the eye there is autophagy but no mitophagy. On the other hand, we have defined that to take place the mitophagy in the ONL, there is a transport from the internal segment to the area of the somas of both lysosomes and mitolysosomes...
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Tesis inédita de la Universidad Complutense de Madrid, Facultad de Ciencias Químicas, Departamento de Bioquímica y Biología Molecular, leída el 29/11/2019