Visualization of gene expression in whole mouse retina by in situ hybridization.

Nature Protocols
Michael B PownerMarcus Fruttiger

Abstract

The mouse retinal vasculature provides a powerful model system for studying development and pathologies of the vasculature. Because it forms as a two-dimensional flat plexus, it is easily imaged in its entirety in whole-mount retinal preparations. In order to study molecular signaling mechanisms, it is useful to visualize the expression of specific genes in the entire vascular plexus and retina. However, in situ hybridization on whole-mount retinal preparations is problematic because isolated retinas have a tendency to curl up during hybridization and are difficult to stain. Here we provide a detailed protocol that overcomes these difficulties and visualizes the mRNA distribution of one or two genes in the context of the counterstained retinal vasculature. The protocol takes 3-4 d for single-probe stains, with an additional 2 d for immunohistochemistry co-labeling. In situ hybridization with two probes adds a further 3 d.

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Citations

Nov 29, 2015·Microcirculation : the Official Journal of the Microcirculatory Society, Inc·Bruce A CorlissWalter Lee Murfee
May 17, 2017·Nature Communications·Do Young ParkGou Young Koh
Feb 15, 2018·International Journal of Molecular Sciences·Mohan NairSiobhan A Corbett
May 17, 2018·Angiogenesis·Patrycja Nowak-SliwinskaArjan W Griffioen
Apr 24, 2020·Diabetes·Bruce A CorlissShayn M Peirce
Jul 29, 2017·Angiogenesis·Henar CuervoChyuan-Sheng Lin
Nov 2, 2019·Scientific Reports·Lauren A LaboissonniereJeffrey M Trimarchi
Aug 28, 2019·The Journal of Clinical Investigation·Nathalie TischCarmen Ruiz de Almodovar
Sep 29, 2018··Tetsuya OkajimaMitsutaka Ogawa

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