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Slit2 is necessary for optic axon organization in the zebrafish ventral midline

View ORCID ProfileCamila Davison, View ORCID ProfileFlavio R. Zolessi
doi: https://doi.org/10.1101/2020.09.25.314062
Camila Davison
Sección Biología Celular, Facultad de Ciencias, Universidad de la República, Iguá 4225, 11400 Montevideo; Institut Pasteur de Montevideo, Mataojo 2020, 11400 Montevideo. Uruguay
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Flavio R. Zolessi
Sección Biología Celular, Facultad de Ciencias, Universidad de la República, Iguá 4225, 11400 Montevideo; Institut Pasteur de Montevideo, Mataojo 2020, 11400 Montevideo. Uruguay
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  • For correspondence: fzolessi@fcien.edu.uy
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ABSTRACT

Slit-Robo signaling has been implicated in regulating several steps of retinal ganglion cell axon guidance, with a central role assigned to Slit2. We report here the phenotypical characterization of a CRISPR-Cas9-generated zebrafish null mutant for this gene, along with a detailed analysis of its expression pattern by WM-FISH. All evident defects in the optic axons in slit2-/- mutants were detected outside the retina, coincident with the major sites of expression at the ventral forebrain, around the developing optic nerve and anterior to the optic chiasm/proximal tract. Anterograde axon tracing experiments in zygotic and maternal-zygotic mutants, as well as morphants, showed the occurrence of axon sorting defects, which appeared mild at the optic nerve level, but more severe in the optic chiasm and the proximal tract. A remarkable sorting defect was the usual splitting of one of the optic nerves in two branches that surrounded the contralateral nerve at the chiasm. Although all axons eventually crossed the midline, the retinotopic order appeared lost at the proximal optic tract, to eventually correct distally. Time-lapse analysis demonstrated the sporadic occurrence of axon misrouting at the chiasm level, which could be responsible for the sorting errors. Our results support previous evidence of a channeling role for Slit molecules in retinal ganglion cell axons at the optic nerve, in addition to a function in the segregation of axons coming from each nerve and from different retinal regions at the medio-ventral area of the forebrain.

Competing Interest Statement

The authors have declared no competing interest.

Footnotes

  • Conflict of interest: The authors declare no conflict of interest.

  • Data availability statement: The data that support the findings of this study, as well as original materials, are available from the corresponding author upon reasonable request.

  • Minor revision of the previous version, including new data regarding expression analysis using RT-PCR at different developmental stages.

  • ABBREVIATIONS

    CRISPR
    clustered regularly interspaced short palindromic repeats
    MO
    morpholino oligomer
    RGC
    retinal ganglion cell
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    Posted March 25, 2021.
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    Slit2 is necessary for optic axon organization in the zebrafish ventral midline
    Camila Davison, Flavio R. Zolessi
    bioRxiv 2020.09.25.314062; doi: https://doi.org/10.1101/2020.09.25.314062
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    Slit2 is necessary for optic axon organization in the zebrafish ventral midline
    Camila Davison, Flavio R. Zolessi
    bioRxiv 2020.09.25.314062; doi: https://doi.org/10.1101/2020.09.25.314062

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