3D-reconstructed retinal pigment epithelial cells provide insights into the anatomy of the outer retina
3D-reconstructed retinal pigment epithelial cells provide insights into the anatomy of the outer retina
The retinal pigment epithelium (RPE) is located between the neuroretina and the choroid, and plays a critical role in vision. RPE cells internalise outer segments (OS) from overlying photoreceptors in the daily photoreceptor renewal. Changes to RPE structure are linked with age and retinopathy, which has been described in the past by conventional 2D electron microscopy. We used serial block face scanning electron microscopy (SBF-SEM) to reconstruct RPE cells from the central mouse retina. Three-dimensional-reconstructed OS revealed the RPE to support large numbers of photoreceptors (90–216 per RPE cell). Larger bi-nucleate RPE maintained more photoreceptors, although their cytoplasmic volume was comparable to smaller mono-nucleate RPE supporting fewer photoreceptors. Scrutiny of RPE microvilli and interdigitating OS revealed the angle and surface area of contact between RPE and photoreceptors. Bi-nucleate RPE contained more mitochondria compared to mono-nucleate RPE. Furthermore, bi-nucleate cells contained larger sub-RPE spaces, supporting a likely association with disease. Use of perfusion-fixed tissues ensured the highest possible standard of preservation, providing novel insights into the 3D RPE architecture and changes linked with retinopathy. This study serves as a benchmark for comparing retinal tissues from donor eyes with age-related macular degeneration (AMD) and other retinopathies.
3D reconstruction, Age‐related macular degeneration (AMD), Imaging, Mouse, Photoreceptors, Retina, Retinal pigment epithelium (RPE), SBF‐SEM
Keeling, Eloise
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Chatelet, David S.
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Tan, Nicole Y. T.
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Khan, Farihah
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Richards, Rhys
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Thisainathan, Thibana
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Goggin, Patricia
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Page, Anton
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Tumbarello, David A.
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Lotery, Andrew J.
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Ratnayaka, J. Arjuna
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9 November 2020
Keeling, Eloise
3207bbdb-d391-44af-8abc-a60c08dce45b
Chatelet, David S.
6371fd7a-e274-4738-9ccb-3dd4dab32928
Tan, Nicole Y. T.
439772f0-08c0-47a9-b3a0-c88f82728de6
Khan, Farihah
cf3520ff-5699-4f25-aaea-4215bab53fbe
Richards, Rhys
d5e04d52-3a2b-4f4a-b45b-72a81e25c94b
Thisainathan, Thibana
b14aed45-0ba1-4ee2-b375-f8c3c078b9a8
Goggin, Patricia
e1e6172b-6e76-47aa-a2a6-3f2a7f4a0a7d
Page, Anton
76ebbfb8-4fe3-495c-afff-1f2f34977fee
Tumbarello, David A.
75c6932e-fdbf-4d3c-bb4f-48fbbdba93a2
Lotery, Andrew J.
5ecc2d2d-d0b4-468f-ad2c-df7156f8e514
Ratnayaka, J. Arjuna
002499b8-1a9f-45b6-9539-5ac145799dfd
Keeling, Eloise, Chatelet, David S., Tan, Nicole Y. T., Khan, Farihah, Richards, Rhys, Thisainathan, Thibana, Goggin, Patricia, Page, Anton, Tumbarello, David A., Lotery, Andrew J. and Ratnayaka, J. Arjuna
(2020)
3D-reconstructed retinal pigment epithelial cells provide insights into the anatomy of the outer retina.
International Journal of Molecular Sciences, 21 (21), [8408].
(doi:10.3390/ijms21218408).
Abstract
The retinal pigment epithelium (RPE) is located between the neuroretina and the choroid, and plays a critical role in vision. RPE cells internalise outer segments (OS) from overlying photoreceptors in the daily photoreceptor renewal. Changes to RPE structure are linked with age and retinopathy, which has been described in the past by conventional 2D electron microscopy. We used serial block face scanning electron microscopy (SBF-SEM) to reconstruct RPE cells from the central mouse retina. Three-dimensional-reconstructed OS revealed the RPE to support large numbers of photoreceptors (90–216 per RPE cell). Larger bi-nucleate RPE maintained more photoreceptors, although their cytoplasmic volume was comparable to smaller mono-nucleate RPE supporting fewer photoreceptors. Scrutiny of RPE microvilli and interdigitating OS revealed the angle and surface area of contact between RPE and photoreceptors. Bi-nucleate RPE contained more mitochondria compared to mono-nucleate RPE. Furthermore, bi-nucleate cells contained larger sub-RPE spaces, supporting a likely association with disease. Use of perfusion-fixed tissues ensured the highest possible standard of preservation, providing novel insights into the 3D RPE architecture and changes linked with retinopathy. This study serves as a benchmark for comparing retinal tissues from donor eyes with age-related macular degeneration (AMD) and other retinopathies.
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Accepted/In Press date: 6 November 2020
e-pub ahead of print date: 9 November 2020
Published date: 9 November 2020
Additional Information:
Funding Information:
Funding: This research was funded by awards to J.A.R. from the UK Macular Society and the Gift of Sight Appeal.
Publisher Copyright:
© 2020 by the authors. Licensee MDPI, Basel, Switzerland.
Keywords:
3D reconstruction, Age‐related macular degeneration (AMD), Imaging, Mouse, Photoreceptors, Retina, Retinal pigment epithelium (RPE), SBF‐SEM
Identifiers
Local EPrints ID: 445269
URI: http://eprints.soton.ac.uk/id/eprint/445269
ISSN: 1422-0067
PURE UUID: 0859907a-e7d2-4b92-8ba7-5c8c98a6b317
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Date deposited: 27 Nov 2020 17:32
Last modified: 17 Mar 2024 03:56
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Contributors
Author:
Eloise Keeling
Author:
David S. Chatelet
Author:
Nicole Y. T. Tan
Author:
Farihah Khan
Author:
Rhys Richards
Author:
Thibana Thisainathan
Author:
Patricia Goggin
Author:
Anton Page
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