June 2021
Volume 62, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2021
A novel enrichment method reveals transcriptional changes in retinal astrocytes exposed to elevated IOP
Author Affiliations & Notes
  • Paul Cullen
    Optometry & Vision Science, University of California Berkeley, Berkeley, California, United States
  • Yujia Yang
    Optometry & Vision Science, University of California Berkeley, Berkeley, California, United States
  • Sandra Muroy
    Optometry & Vision Science, University of California Berkeley, Berkeley, California, United States
  • Lu Chen
    Optometry & Vision Science, University of California Berkeley, Berkeley, California, United States
  • John G Flanagan
    Optometry & Vision Science, University of California Berkeley, Berkeley, California, United States
  • Footnotes
    Commercial Relationships   Paul Cullen, None; Yujia Yang, None; Sandra Muroy, None; Lu Chen, None; John Flanagan, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science June 2021, Vol.62, 1701. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Paul Cullen, Yujia Yang, Sandra Muroy, Lu Chen, John G Flanagan; A novel enrichment method reveals transcriptional changes in retinal astrocytes exposed to elevated IOP. Invest. Ophthalmol. Vis. Sci. 2021;62(8):1701.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose : The neuroprotective role of glial cells is a growing research focus in nervous system pathologies including glaucoma, which is one of the most common neurodegenerative disorders and the leading cause of irreversible blindness worldwide. Retinal astrocytes, along with those of the optic nerve, have been shown to play key roles in the survival of retinal ganglion cells, particularly in response to glaucomatous stress. The mechanisms regulating this protection, however, are poorly understood, as these cells populate the retina sparsely and are difficult to isolate acutely. By utilizing a novel isolation method and examining the transcriptomic data from retinal astrocytes exposed to elevated intraocular pressure (IOP), we aim to understand the response of these cells to glaucomatous stress.

Methods : Adult male C57/B6 mice underwent laser-induced episcleral vein cauterization, resulting in an elevation in IOP. After one week, treated animals and age-matched controls were euthanized and highly enriched populations of retinal astrocytes were isolated by a combination of enzymatic dissociation and mechanical separation. Purified RNA from these samples was used to generate cDNA libraries for next generation sequencing, and differential gene expression was analyzed after bioinformatic cleanup.

Results : After applying thresholds of 2-fold change in expression and <.05 for false discovery rate, 335 upregulated genes were identified in samples from experimental animals relative to those from controls. The most strongly correlated gene ontology category was 'GO:0006954: inflammatory response', with a -log10(P) of 27.8. Furthermore, a number of these upregulated genes have been previously implicated as markers of astrocyte reactivity. The second most strongly correlated GO category was 0001568: blood vessel development, with a -log10(P) of 25.1, likely reflecting the close association of astrocytes with the superficial retinal vasculature.

Conclusions : Our results are broadly consistent with studies of reactive astrocytes in the brain, while delivering new insight into the behavior of these cells in the retina; this may illuminate future directions for research into retinal neuroprotection. This outcome also demonstrates the utility of our approach to retinal astrocyte isolation, which requires little in the way of specialized equipment and benefits from a post-mortem interval of less than one hour.

This is a 2021 ARVO Annual Meeting abstract.

×
×

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.

×