June 2015
Volume 56, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2015
Rod Photoreceptor-Enriched Transcripts of Zebrafish Retina
Author Affiliations & Notes
  • CHI SUN
    Biological Sciences, University of Idaho, Moscow, ID
  • Deborah L Stenkamp
    Biological Sciences, University of Idaho, Moscow, ID
  • Footnotes
    Commercial Relationships CHI SUN, None; Deborah Stenkamp, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2015, Vol.56, 5493. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      CHI SUN, Deborah L Stenkamp; Rod Photoreceptor-Enriched Transcripts of Zebrafish Retina. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):5493.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
Abstract

Purpose: The purpose of the current study is to identify transcripts that are enriched in rod photoreceptors, in order to gain insights into intrinsic mechanisms underlying rod development and survival.

Methods: We use a transgenic zebrafish line (XOPS:eGFP) in which rod photoreceptors express green fluorescent protein (GFP) as a model organism for this study. Enzymatic dissociation of adult retinal tissue successfully separated individual cells. Rod photoreceptors were isolated using a BD Biosciences FACS Aria flow cytometer, based on their expression of GFP. High-quality RNA samples were subsequently amplified and used to construct eight cDNA libraries (4 GFP+ and 4 GFP-) based on samples from four fish, and these were subjected to RNA-seq.

Results: Sorted GFP+ (rods) cells constituted 10-20% of all retinal cells from a single fish. Approximately 150,000 to 250,000 events were collected in each GFP+ fraction. The list of differentially expressed (GFP+ vs. GFP-) transcripts with FDR < 0.01 consisted of 615 entries. These included known rod-specific transcripts such as rhodopsin and nr2e3, as well as transcripts not previously known to be enriched in rods. In addition, several transcripts corresponding to mitochondrial enzymes were enriched in rods.

Conclusions: RNA-seq and subsequent analysis identified numerous transcripts differentially expressed in rod photoreceptors as compared to other retinal cells. qRT-PCR and in situ hybridization studies of selected transcripts are underway to verify differential expression and identify rod markers.

×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×