June 2017
Volume 58, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2017
Title: Characterization of Capn5 expression and function in the zebrafish retina
Author Affiliations & Notes
  • Cagney Coomer
    Biology, University of Kentucky, Lexington, Kentucky, United States
  • Ann C Morris
    Biology, University of Kentucky, Lexington, Kentucky, United States
  • Footnotes
    Commercial Relationships   Cagney Coomer, None; Ann Morris, None
  • Footnotes
    Support  NIH Grant EY021769, Lyman T. Johnson Fellowship
Investigative Ophthalmology & Visual Science June 2017, Vol.58, 117. doi:
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      Cagney Coomer, Ann C Morris; Title: Characterization of Capn5 expression and function in the zebrafish retina
      . Invest. Ophthalmol. Vis. Sci. 2017;58(8):117.

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

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Abstract

Purpose : Purpose: Autosomal dominant neovascular inflammatory vitreoretinopathy (ADNIV) is a devastating inherited autoimmune disease of the eye that displays features commonly seen in other eye diseases, such as retinitis pigmentosa and diabetic retinopathy. A gain of function mutation in Calpain-5 (capn5), a calcium dependent cysteine protease, has been implicated as the cause of ADNIV. Very little is known about the normal function of capn5 in the adult retina, and there are conflicting results regarding its role during embryonic development. In this study, we characterized the expression of capn5 in the developing and adult zebrafish retina as well as during photoreceptor degeneration and regeneration.

Methods : Methods: All animal procedures were performed in accordance with guidelines established by the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research. The XOPS:mCFP and TaC:GFP transgenic lines have been previously described. Zebrafish embryos were staged and collected at various time points encompassing retinal development. Expression of capn5 was analyzed by rt-PCR, qPCR, whole mount in situ hybridization (WISH) and fluorescent in situ hybridization (FISH). Light damage procedures were performed as previously described.

Results : Results: We found that capn5 is expressed in the central nervous system (CNS) during embryonic development, and we observed expression in the outer nuclear layer (ONL) of the developing retina starting at 3 dpf. Expression of capn5 co-localized with expression of cone specific genes in the adult zebrafish retina. We observed an increase in expression of capn5 in the XOPs-mCFP transgenic zebrafish retina, which experiences chronic rod photoreceptor degeneration and regeneration, compared to wild type controls. Acute light damage to the zebrafish retina resulted in photoreceptor degeneration, which was accompanied by an increase in expression of capn5.

Conclusions : Conclusions: Our results demonstrate that capn5 is expressed in the CNS and retina of the adult and developing zebrafish. This expression is possibly cone specific in the retina and is upregulated in response to rod photoreceptor degeneration. These studies suggest that Capn5 may play a role in photoreceptor development and maintenance.

This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.

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