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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
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, Retina, SBF-SEM, photoreceptors, retinal pigment epithelium (RPE)
1422-0067
1
Keeling, Eloise
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Chatelet, David S.
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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
c2d225c4-b84d-4ead-a433-320057cb5fa9
Page, Anton
76ebbfb8-4fe3-495c-afff-1f2f34977fee
Tumbarello, David A.
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Lotery, Andrew J.
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Ratnayaka, J. Arjuna
002499b8-1a9f-45b6-9539-5ac145799dfd
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
c2d225c4-b84d-4ead-a433-320057cb5fa9
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), 1, [8408]. (doi:10.3390/ijms21218408).

Record type: Article

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
Keywords: 3D reconstruction, Age-related Macular Degeneration (AMD), Imaging, Mouse, Retina, SBF-SEM, photoreceptors, retinal pigment epithelium (RPE)

Identifiers

Local EPrints ID: 445269
URI: http://eprints.soton.ac.uk/id/eprint/445269
ISSN: 1422-0067
PURE UUID: 0859907a-e7d2-4b92-8ba7-5c8c98a6b317
ORCID for Patricia Goggin: ORCID iD orcid.org/0000-0003-4730-0206
ORCID for David A. Tumbarello: ORCID iD orcid.org/0000-0002-5169-0561
ORCID for Andrew J. Lotery: ORCID iD orcid.org/0000-0001-5541-4305
ORCID for J. Arjuna Ratnayaka: ORCID iD orcid.org/0000-0002-1027-6938

Catalogue record

Date deposited: 27 Nov 2020 17:32
Last modified: 26 Nov 2021 03:08

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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 ORCID iD
Author: Anton Page

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