An easy, rapid method to isolate RPE cell protein from the mouse eye

Hong Wei, Zixian Xun, Herta Granado, Angela Wu, James T. Handa

Research output: Contribution to journalShort surveypeer-review

Abstract

The retinal pigment epithelium (RPE) is essential for maintaining the health of the neural retina. RPE cell dysfunction plays a critical role in many common blinding diseases including age-related macular degeneration (AMD), diabetic retinopathy, retinal dystrophies. Mouse models of ocular disease are commonly used to study these blinding diseases. Since isolating the RPE from the choroid has been challenging, most techniques separate the RPE from the retina, but not the choroid. As a result, the protein signature actually represents a heterogeneous population of cells that may not accurately represent the RPE response. Herein, we describe a method for separating proteins from the RPE that is free from retinal and choroidal contamination. After removing the anterior segment and retina from enucleated mouse eyes, protein from the RPE was extracted separately from the choroid by incubating the posterior eyecup with a protein lysis buffer for 10 min. Western blot analysis identified RPE65, an RPE specific protein in the RPE lysates, but not in choroidal lysates. The RPE lysates were devoid of rhodopsin and collagen VI, which are abundant in the retina and choroid, respectively. This technique will be very helpful for measuring the protein signal from the RPE without retinal or choroidal contamination.

Original languageEnglish (US)
Pages (from-to)450-455
Number of pages6
JournalExperimental eye research
Volume145
DOIs
StatePublished - Apr 1 2016

Keywords

  • Choroid
  • Dissection
  • Mouse
  • Retinal pigmented epithelium

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Fingerprint Dive into the research topics of 'An easy, rapid method to isolate RPE cell protein from the mouse eye'. Together they form a unique fingerprint.

Cite this