June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
Functional analysis of a zebrafish knockdown of the optic atrophy-associated gene ssbp1
Author Affiliations & Notes
  • Julian Perrin
    INSERM U1298, Institut des Neurosciences de Montpellier, Montpellier, Occitanie, France
  • Vincent Gisbert
    INSERM U1298, Institut des Neurosciences de Montpellier, Montpellier, Occitanie, France
  • Nicolas Cubedo
    INSERM U1198, Mecanismes moleculaires dans les demences neurodegeneratives, Montpellier, Occitanie, France
  • Mireille Rossel
    INSERM U1198, Mecanismes moleculaires dans les demences neurodegeneratives, Montpellier, Occitanie, France
  • Marie Pequignot
    INSERM U1298, Institut des Neurosciences de Montpellier, Montpellier, Occitanie, France
  • Cécile Delettre
    INSERM U1298, Institut des Neurosciences de Montpellier, Montpellier, Occitanie, France
  • Footnotes
    Commercial Relationships   Julian Perrin None; Vincent Gisbert None; Nicolas Cubedo None; Mireille Rossel None; Marie Pequignot None; Cécile Delettre None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 977 – F0374. doi:
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      Julian Perrin, Vincent Gisbert, Nicolas Cubedo, Mireille Rossel, Marie Pequignot, Cécile Delettre; Functional analysis of a zebrafish knockdown of the optic atrophy-associated gene ssbp1. Invest. Ophthalmol. Vis. Sci. 2022;63(7):977 – F0374.

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

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Abstract

Purpose : Dominant Optic Atrophy (DOA) is characterized by degeneration of the retinal ganglion cells (RGC) that can lead to vision loss for which there is no treatment. The optic atrophy (OA) can be isolated or syndromic: OA associated with sensorineural deafness, myopathy, etc. DOA is caused by mutations in genes with a mitochondrial function. The underlying pathophysiology remains to be understood.
The single strand DNA-binding protein 1, SSBP1 gene was recently identified for causing DOA. SSBP1 tetramers bind mitochondrial DNA (mtDNA) and is essential for mtDNA replication. Our purpose is to shed light on the pathophysiological consequences of ssbp1 alteration in a zebrafish model.

Methods : A knockdown (KD) of ssbp1 in zebrafish was generated by microinjection of translation blocking morpholino (MO-ssbp1) in Zebrafish embryo. Fish injected with Standard-MO designed to have no impact in zebrafish and non-injected zebrafish were used as controls. KD was validated by Western Blot analysis of Ssbp1.
The transgenic zebrafish line tg(Brn3c:mGFP) labeling RGC and ON was used throughout this study. Confocal microscopy and the IMARIS software were used to generate 3D reconstruction of the optic nerve GFP labeling.
Sensorimotor responses were investigated using either Visual Motor Response assay (VMR) to measure the locomotor response to a visual stimulus (traveled distance measurement) or Auditory Startle Response (ASR) to measure motility after acoustic and vibration stimuli.

Results : ON morphology analysis revealed an atrophy of the ON in 2dpf ssbp1 KD zebrafish, with a significant reduction of its volume.
A significant decrease of the traveled distanced by 5dpf MO-ssbp1 fish was found in both light and dark stimuli conditions using VMR assay. However, the ability to perceive differences in bright light stimulation did not seem impaired in MO-ssbp1.
A diminution of motility of 5dpf MO-ssbp1 was also found using the ASR assay and preliminary result show a decrease of startle response induced by sound and vibration stimuli.

Conclusions : ssbp1 KD in zebrafish revealed an optic nerve atrophy at 2 dpf which could indicate that ssbp1 is important for ON development. This result did not translate to bright light perception defect in 5dpf ssbp1 KD fish. Moreover, the KD caused an overall diminution of the traveled distance that could reflect locomotion defect as well as a likely alteration of sound and vibration perception.

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

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