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Study of intracellular cargo trafficking and co-localization in the phagosome and autophagy-lysosomal pathways of Retinal Pigment Epithelium (RPE) cells

Study of intracellular cargo trafficking and co-localization in the phagosome and autophagy-lysosomal pathways of Retinal Pigment Epithelium (RPE) cells
Study of intracellular cargo trafficking and co-localization in the phagosome and autophagy-lysosomal pathways of Retinal Pigment Epithelium (RPE) cells
The transport and targeting of internalized molecules to distinct intracellular organelles/compartments can prove challenging to visualize clearly, which can contribute to some of the difficulties associated with these studies. By combining several approaches, we show how the trafficking and processing of photoreceptor outer segments in the phagosome and autophagy-lysosomal pathways of the retinal pigment epithelium (RPE) can easily be quantified and visualized as 3D-reconstructed images. This protocol takes advantage of new developments in microscopy and image-analysis software which has the potential to help better understand dynamic intracellular processes that underlie RPE dysfunction associated with irreversible blinding diseases such as age-related macular degeneration. The method described herein can also be used to study the trafficking and co-localization of different intracellular cargos in other cell types and tissues.
3D imaging, Co-localization, Retinal pigment epithelium (RPE), Phagosome, Lysosomes, Autophagy, Confocal Microscopy
Springer
Ratnayaka, J. Arjuna
002499b8-1a9f-45b6-9539-5ac145799dfd
Keeling, Eloise
abc30700-b1ed-49ed-ba1c-078856e01b7c
Chatelet, David
6371fd7a-e274-4738-9ccb-3dd4dab32928
Turksen, Kursad
Ratnayaka, J. Arjuna
002499b8-1a9f-45b6-9539-5ac145799dfd
Keeling, Eloise
abc30700-b1ed-49ed-ba1c-078856e01b7c
Chatelet, David
6371fd7a-e274-4738-9ccb-3dd4dab32928
Turksen, Kursad

Ratnayaka, J. Arjuna, Keeling, Eloise and Chatelet, David (2019) Study of intracellular cargo trafficking and co-localization in the phagosome and autophagy-lysosomal pathways of Retinal Pigment Epithelium (RPE) cells. In, Turksen, Kursad (ed.) Methods in Molecular Biology: Imaging and Tracking Stem Cells. (Methods in Molecular Biology) Cham. Springer. (doi:10.1007/7651_2019_223).

Record type: Book Section

Abstract

The transport and targeting of internalized molecules to distinct intracellular organelles/compartments can prove challenging to visualize clearly, which can contribute to some of the difficulties associated with these studies. By combining several approaches, we show how the trafficking and processing of photoreceptor outer segments in the phagosome and autophagy-lysosomal pathways of the retinal pigment epithelium (RPE) can easily be quantified and visualized as 3D-reconstructed images. This protocol takes advantage of new developments in microscopy and image-analysis software which has the potential to help better understand dynamic intracellular processes that underlie RPE dysfunction associated with irreversible blinding diseases such as age-related macular degeneration. The method described herein can also be used to study the trafficking and co-localization of different intracellular cargos in other cell types and tissues.

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Accepted/In Press date: 2019
Published date: 11 April 2019
Keywords: 3D imaging, Co-localization, Retinal pigment epithelium (RPE), Phagosome, Lysosomes, Autophagy, Confocal Microscopy

Identifiers

Local EPrints ID: 429955
URI: http://eprints.soton.ac.uk/id/eprint/429955
PURE UUID: 9d58f631-7ee9-40ee-832b-15909ee6ba21
ORCID for J. Arjuna Ratnayaka: ORCID iD orcid.org/0000-0002-1027-6938

Catalogue record

Date deposited: 15 Apr 2019 16:30
Last modified: 26 Nov 2021 03:01

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Contributors

Author: Eloise Keeling
Author: David Chatelet
Editor: Kursad Turksen

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