Retinal Layer Separation (ReLayS) method enables the molecular analysis of photoreceptor segments and cell bodies, as well as the inner retina

Understanding the physiology of the retina, and especially of the highly polarized photoreceptors, is essential not only to broaden our knowledge of the processes required for normal vision, but also to develop effective therapies to prevent or slow retinal degenerative diseases. However, the molecu...

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Bibliographic Details
Published in:	Scientific reports Vol. 12; no. 1; pp. 20195 - 10
Main Authors:	Todorova, Vyara, Merolla, Luca, Karademir, Duygu, Wögenstein, Gabriele M., Behr, Julian, Ebner, Lynn J. A., Samardzija, Marijana, Grimm, Christian
Format:	Journal Article
Language:	English
Published:	London Nature Publishing Group UK 23-11-2022 Nature Publishing Group Nature Portfolio
Subjects:	631/1647/2230 631/378/2613/1786 Aging Animals Cell Body - metabolism Degenerative diseases Heterogeneity Humanities and Social Sciences Metabolites Mice multidisciplinary Photoreception Photoreceptor Cells - metabolism Photoreceptors Retina Retina - metabolism Retinal degeneration Science Science (multidisciplinary) Transducin Transducin - metabolism Translocation Western blotting
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Summary:	Understanding the physiology of the retina, and especially of the highly polarized photoreceptors, is essential not only to broaden our knowledge of the processes required for normal vision, but also to develop effective therapies to prevent or slow retinal degenerative diseases. However, the molecular analysis of photoreceptors is a challenge due to the heterogeneity of the retinal tissue and the lack of easy and reliable methods for cell separation. Here we present the ReLayS method—a simple technique for the separation of photoreceptor segments (PS) containing both inner and outer segments, outer nuclear layer (ONL), and inner retina (InR) that contains the remaining retinal layers. The layer-specific material isolated from a mouse half-retina with the ReLayS method was sufficient for protein isolation and Western blotting or RNA isolation and real-time PCR studies. The separation of PS, ONL, and InR was successfully validated by Western blotting and real-time PCR using proteins and genes with known expression profiles within the retina. Furthermore, the separation of the PS from the ONL enabled the detection of light-driven translocation of transducin from the PS to the soma. ReLayS is a simple and useful method to address protein and possibly metabolites distribution in photoreceptor compartments in various situations including development, ageing, and degenerative diseases.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	2045-2322 2045-2322
DOI:	10.1038/s41598-022-24586-8