September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
Genetic ablation of miR-129-3p in zebrafish does not affect cone outer segment development
Author Affiliations & Notes
  • Joseph Fogerty
    Department of Ophthalmology, Cleveland Clinic Foundation, Cleveland, Ohio, United States
  • Brian D Perkins
    Department of Ophthalmology, Cleveland Clinic Foundation, Cleveland, Ohio, United States
  • Footnotes
    Commercial Relationships   Joseph Fogerty, None; Brian Perkins, None
  • Footnotes
    Support  F32-EY025145, R01-EY017037
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 1773. doi:
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      Joseph Fogerty, Brian D Perkins; Genetic ablation of miR-129-3p in zebrafish does not affect cone outer segment development. Invest. Ophthalmol. Vis. Sci. 2016;57(12):1773.

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

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Abstract

Purpose : MicroRNAs (miRs) function as important regulators of photoreceptor development and physiology. The purpose of this study was to determine if photoreceptor sensory cilium development requires miR-129-3p, a key regulator of ciliogenesis.

Methods : Both morpholinos and TALENs were used to inactivate miR-129-3p in zebrafish. Zebrafish contain two mir-129 paralogs, mir-129-1 and mir-129-2. We designed TALENs against each locus and selected mutations that are predicted to inactivate the seed sequence of each miR-129-3p paralog without significantly altering the pre-miRNA hairpin structure. We used PNA to examine cone outer segment length in 5dpf larvae of both single and double mir-129 mutants.

Results : At 5 days post fertilization (dpf), zebrafish larvae injected with morpholinos targeting miR-129-3p exhibited phenotypes consistent with malfunctioning cilia, including shortened photoreceptor outer segments, kidney cysts, ectopic otoliths, and severe ventral body curvature. Mir-129-1-3p-/-; mir-129-2-3p-/- fish, which are predicted to lack all functional miR-129-3p, were viable and were produced in the expected Mendelian ratio. At 5 dpf, these mutant animals lacked the severe body curvature and kidney cysts that were readily apparent in morphant fish, and PNA staining showed normal cone outer segment lengths.

Conclusions : Contrary to our hypothesis and in stark contrast to miR-129-3p morphant zebrafish, miR-129-3p mutants have no overt cilia phenotypes at 5 dpf. This suggests that the morpholino may non-selectively target additional RNAs that can compensate for loss of miR-129-3p in mutant fish.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

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