June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
Role of 5-hydroxymethylcytosine during postnatal retinal development
Stylianos Michalakis; Arshan Perera; Susanne Koch; Mirko Wagner; Lukas Windhager; Kerstin Nagel-Wolfrum; Tim Strom; Ralf Zimmer; Thomas Carell; Martin Biel
Author Affiliations & Notes
  • Stylianos Michalakis
    Center for Integrated Protein Science at the Department of Pharmacy-Center for Drug Research, Ludwig-Maximilians-Universität München, Munich, Germany
  • Arshan Perera
    Center for Integrated Protein Science at the Department of Pharmacy-Center for Drug Research, Ludwig-Maximilians-Universität München, Munich, Germany
  • Susanne Koch
    Center for Integrated Protein Science at the Department of Pharmacy-Center for Drug Research, Ludwig-Maximilians-Universität München, Munich, Germany
  • Mirko Wagner
    Center for Integrated Protein Science at the Department of Chemistry, Ludwig-Maximilians-Universität München, Munich, Germany
  • Lukas Windhager
    Institute for Informatics, Ludwig-Maximilians-Universität München, Munich, Germany
  • Kerstin Nagel-Wolfrum
    Department of Cell and Matrix Biology, Institute of Zoology, Johannes Gutenberg University-Mainz, Mainz, Germany
  • Tim Strom
    Institute of Human Genetics, Technische Universität München, Munich, Germany
    Institute of Human Genetics, Helmholtz Zentrum München, German Research Center for Environmental Health, Neuherberg, Germany
  • Ralf Zimmer
    Institute for Informatics, Ludwig-Maximilians-Universität München, Munich, Germany
  • Thomas Carell
    Center for Integrated Protein Science at the Department of Chemistry, Ludwig-Maximilians-Universität München, Munich, Germany
  • Martin Biel
    Center for Integrated Protein Science at the Department of Pharmacy-Center for Drug Research, Ludwig-Maximilians-Universität München, Munich, Germany
  • Footnotes
    Commercial Relationships Stylianos Michalakis, None; Arshan Perera, None; Susanne Koch, None; Mirko Wagner, None; Lukas Windhager, None; Kerstin Nagel-Wolfrum, None; Tim Strom, None; Ralf Zimmer, None; Thomas Carell, None; Martin Biel, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 2618. doi:
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      Stylianos Michalakis, Arshan Perera, Susanne Koch, Mirko Wagner, Lukas Windhager, Kerstin Nagel-Wolfrum, Tim Strom, Ralf Zimmer, Thomas Carell, Martin Biel; Role of 5-hydroxymethylcytosine during postnatal retinal development. Invest. Ophthalmol. Vis. Sci. 2013;54(15):2618.

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Abstract

Purpose: 5-hydroxymethylcytosine (5hmC), also known as the sixth base of the genome, is a recently discovered oxidative product of 5-methylcytosine (5mC) generated by the enzymatic action of Ten Eleven Translocation (TET) family members. The functional role of 5hmC is largly unknown. However, studies in embryonic stem cells and cancer tissues suggest that TET enzymes and 5hmC may be involved in gene regulation. Eye opening at postnatal week (pw) 2 is a key time point during mouse retinal development. Maturation of retinal cells as well as formation of the retinal network is ongoing after eye opening and is completed only one to two weeks later. Here, we analyzed the role of 5hmC during this phase of postnatal retinal development in the mouse.

Methods: 5hmC was localized to specific retinal cell types using immunohistochemistry. Global 5hmC levels in retinal samples were quantified using ultra high pressure liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). 5hmC marks were mapped to the retinal genome by hydroxymethylated DNA immunoprecipitation (hMeDIP) followed by next generation sequencing and were subsequently correlated with retinal gene expression data obtained from microarray experiments.

Results: At pw 2 5hmC weakly localized to nuclei of cells within the ganglion cell layer and the inner nuclear layer of the retina. The levels of 5hmC in these cells were increased at pw 3. In addition, 5hmC was now detectable in retinal photoreceptors were it co-localized with histone marks of the euchromatin. Global levels of 5hmC also increased from pw 2 to pw 3, whereas 5-methylcytosine was unchanged. hMeDIP experiments revealed a developmentally programmed acquisition of 5hmC during retinal maturation at gene-rich regions and in genes containing activating histone marks.

Conclusions: Our data suggest that 5hmC is dynamically regulated during postnatal retinal development and is capable to elevate gene expression of retina-specific genes.

Keywords: 698 retinal development • 536 gene modifiers • 533 gene/expression  
© 2013, The Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Permission to republish any abstract or part of an abstract in any form must be obtained in writing from the ARVO Office prior to publication.
Copyright © 2025 Association for Research in Vision and Ophthalmology.
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