June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
Cell-specific Studies Through Novel Mouse Model Reveal the Transcriptomic Response of Retinal Müller Glia to Aging
Author Affiliations & Notes
  • Ana J Chucair-Elliott
    Genes & Human Disease, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, United States
  • Sarah Ocañas
    Genes & Human Disease, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, United States
    Physiology, The University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, United States
  • Kevin Pham
    Genes & Human Disease, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, United States
  • Michael Van Der Veldt
    Genes & Human Disease, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, United States
  • Ashley Cheyney
    Physiology, The University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, United States
  • David Stanford
    Genes & Human Disease, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, United States
  • Michael H Elliott
    Ophthalmology, The University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, United States
    Physiology, The University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, United States
  • Willard Freeman
    Genes & Human Disease, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, United States
    Oklahoma City VA Medical Center, Oklahoma City, Oklahoma, United States
  • Footnotes
    Commercial Relationships   Ana Chucair-Elliott None; Sarah Ocañas None; Kevin Pham None; Michael Van Der Veldt None; Ashley Cheyney None; David Stanford None; Michael Elliott None; Willard Freeman None
  • Footnotes
    Support  This work was supported by grants from BrightFocus Foundation (M2020207), the National Institutes of Health (NIH) P30AG050911, R56AG059430, R01AG059430, P20GM125528, T32AG052363, F31AG064861, P30EY021725, R01AG052606, Oklahoma Center for Adult Stem Cell Research (OCASCR), a program of the Oklahoma Tobacco Settlement Endowment Trust, and Presbyterian Health Foundation. This work was also supported in part by the MERIT award I01BX003906 from the United States (U.S.) Department of Veterans Affairs, Biomedical Laboratory Research and Development Service.
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 3483. doi:
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    • Get Citation

      Ana J Chucair-Elliott, Sarah Ocañas, Kevin Pham, Michael Van Der Veldt, Ashley Cheyney, David Stanford, Michael H Elliott, Willard Freeman; Cell-specific Studies Through Novel Mouse Model Reveal the Transcriptomic Response of Retinal Müller Glia to Aging. Invest. Ophthalmol. Vis. Sci. 2022;63(7):3483.

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

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Abstract

Purpose : Aging is the major risk factor for age-related macular degeneration (AMD) but how aging contributes to AMD pathogenesis is unclear. Epigenetic modifications, mainly methylcytosine (mC) and hydroxymethylcytosine (hmC), regulate the genome and gene expression, and are altered with advanced age. DNA methylation analyses of AMD patient blood and retinas suggest differential methylation resulting in gene expression changes during development of AMD. In addition, Müller glial cells (MGC) are activated in AMD patient retinas. The aim of this study is the validation and demonstration of a cre/ERT2-NuTRAP mouse model for the paired interrogation of differential mC/hmC genome-wide and the translatome specifically in MGC with retinal aging.

Methods : Female NuTRAP flox/flox and male Aldh1l1-cre/ERT2+/wt mice were bred to generate Aldh1l1-cre/ERT2+/wt; NuTRAPflox/wt (Aldh1l1-cre/ERT2+; NuTRAP+) mice. After systemic tamoxifen treatment mice were euthanized at 6 or 24 months of age and their retinae harvested and processed for: immunohistochemistry of mCherry, EGFP, GS, CD11b, and GFAP expression in retina sagittal sections via confocal microscopy; MGC ribosome bound RNA isolation via TRAP and stranded RNAseq profile; MGC nuclei isolation via INTACT method for bisulfite amplicon sequencing (BSAS) and nuclear RNAseq.

Results : Specific co-localization of EGFP, mCherry, and the MGC marker GS was observed in the Aldh1l1-cre/ERT2+; NuTRAP+ retinae. RNA-seq bioinformatic analysis revealed MGC-specific transcriptomic changes associated to inflammation in response to aging, such as purinergic receptor and cytokine-mediated signaling pathways. Nuclear RNAseq data supported the MGC identity of the INTACT-DNA isolates. BSAS showed site-specific decrease of mC in the promoter region of the MGC-specific gene Kcnj10 and hypermethylation in the promoter region of the photoreceptor gene Rho in INTACT-DNA positive fraction relative to input.

Conclusions : Our data show the Aldh1l1-NuTRAP model as a suitable tool for the paired interrogation of DNA methylation and gene expression, specifically in MGC. Differential transcript expression with overrepresentation of pathways and processes related to inflammation was found specifically in MGC in response to aging.

This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.

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