Identifying Ciliary Proteins in Mammalian Retinas using a Gentle Extraction Method

TL;DR

This paper introduces a new method to isolate primary cilia from retinas, helping to better understand ciliary proteins and their role in diseases like blindness.

Contribution

A novel gentle extraction method for isolating primary cilia from bovine retinas is developed and validated.

Findings

The method successfully isolates primary cilia from photoreceptor cells.

LC/MS/MS analysis identified proteins enriched for cilia function and ciliopathy disease.

The approach yields sufficient native protein samples for proteomic studies.

Abstract

Mutations in retinal primary cilia are responsible for human blindness but the mechanisms are not fully understood (Wheway et al., 2014). Characterizing the proteome of an organelle such as cilia, is a fruitful way to understand its function but methods often require considerable sample quantities. Here we develop a method to isolate the primary cilia of photoreceptor cells from bovine retinas. Through LC/MS/MS proteomics analysis we identify proteins enriched for cilia function including ciliopathy disease. This study shows our method can be used to isolate retinal primary cilia to obtain sufficient quantities of native protein samples.

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