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661W photoreceptor cell line as a cell model for studying retinal ciliopathies

661W photoreceptor cell line as a cell model for studying retinal ciliopathies
661W photoreceptor cell line as a cell model for studying retinal ciliopathies
The retina contains several ciliated cell types, including the retinal pigment epithelium (RPE) and photoreceptor cells. The photoreceptor cilium is one of the most highly modified sensory cilia in the human body. The outer segment of the photoreceptor is a highly elaborate primary cilium, containing stacks or folds of membrane where the photopigment molecules are located. Perhaps unsurprisingly, defects in cilia often lead to retinal phenotypes, either as part of syndromic conditions involving other organs, or in isolation in the so-called retinal ciliopathies. The study of retinal ciliopathies has been limited by a lack of retinal cell lines. RPE1 retinal pigment epithelial cell line is commonly used in such studies, but the existence of a photoreceptor cell line has largely been neglected in the retinal ciliopathy field. 661W cone photoreceptor cells, derived from mouse, have been widely used as a model for studying macular degeneration, but not described as a model for studying retinal ciliopathies such as retinitis pigmentosa. Here, we characterize the 661W cell line as a model for studying retinal ciliopathies. We fully characterize the expression profile of these cells, using whole transcriptome RNA sequencing, and provide this data on Gene Expression Omnibus for the advantage of the scientific community. We show that these cells express the majority of markers of cone cell origin. Using immunostaining and confocal microscopy, alongside scanning electron microscopy, we show that these cells grow long primary cilia, reminiscent of photoreceptor outer segments, and localize many cilium proteins to the axoneme, membrane and transition zone. We show that siRNA knockdown of cilia genes Ift88 results in loss of cilia, and that this can be assayed by high-throughput screening. We present evidence that the 661W cell line is a useful cell model for studying retinal ciliopathies.
1664-8021
Wheway, Gabrielle
2e547e5d-b921-4243-a071-2208fd4cc090
Nazlamova, Liliya
0cc21013-aeeb-4eef-af56-31f6fa0766fd
Turner, Dann
4be8b141-eb27-472f-8f30-5a176dc0abb9
Cross, Stephen
753150a6-5d23-43be-8eca-47867049c2d6
Wheway, Gabrielle
2e547e5d-b921-4243-a071-2208fd4cc090
Nazlamova, Liliya
0cc21013-aeeb-4eef-af56-31f6fa0766fd
Turner, Dann
4be8b141-eb27-472f-8f30-5a176dc0abb9
Cross, Stephen
753150a6-5d23-43be-8eca-47867049c2d6

Wheway, Gabrielle, Nazlamova, Liliya, Turner, Dann and Cross, Stephen (2019) 661W photoreceptor cell line as a cell model for studying retinal ciliopathies. Frontiers in Genetics. (doi:10.3389/fgene.2019.00308).

Record type: Article

Abstract

The retina contains several ciliated cell types, including the retinal pigment epithelium (RPE) and photoreceptor cells. The photoreceptor cilium is one of the most highly modified sensory cilia in the human body. The outer segment of the photoreceptor is a highly elaborate primary cilium, containing stacks or folds of membrane where the photopigment molecules are located. Perhaps unsurprisingly, defects in cilia often lead to retinal phenotypes, either as part of syndromic conditions involving other organs, or in isolation in the so-called retinal ciliopathies. The study of retinal ciliopathies has been limited by a lack of retinal cell lines. RPE1 retinal pigment epithelial cell line is commonly used in such studies, but the existence of a photoreceptor cell line has largely been neglected in the retinal ciliopathy field. 661W cone photoreceptor cells, derived from mouse, have been widely used as a model for studying macular degeneration, but not described as a model for studying retinal ciliopathies such as retinitis pigmentosa. Here, we characterize the 661W cell line as a model for studying retinal ciliopathies. We fully characterize the expression profile of these cells, using whole transcriptome RNA sequencing, and provide this data on Gene Expression Omnibus for the advantage of the scientific community. We show that these cells express the majority of markers of cone cell origin. Using immunostaining and confocal microscopy, alongside scanning electron microscopy, we show that these cells grow long primary cilia, reminiscent of photoreceptor outer segments, and localize many cilium proteins to the axoneme, membrane and transition zone. We show that siRNA knockdown of cilia genes Ift88 results in loss of cilia, and that this can be assayed by high-throughput screening. We present evidence that the 661W cell line is a useful cell model for studying retinal ciliopathies.

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Accepted/In Press date: 21 March 2019
Published date: 5 April 2019

Identifiers

Local EPrints ID: 430277
URI: https://eprints.soton.ac.uk/id/eprint/430277
ISSN: 1664-8021
PURE UUID: 700d8495-9d2f-45bd-9186-d4eb284fab1e
ORCID for Gabrielle Wheway: ORCID iD orcid.org/0000-0002-0494-0783

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Date deposited: 24 Apr 2019 16:30
Last modified: 25 Apr 2019 00:21

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