1. Developmental Biology
  2. Neuroscience
Download icon

Identification of compounds that rescue otic and myelination defects in the zebrafish adgrg6 (gpr126) mutant

Research Article
  • Cited 9
  • Views 2,044
  • Annotations
Cite this article as: eLife 2019;8:e44889 doi: 10.7554/eLife.44889

Abstract

Adgrg6 (Gpr126) is an adhesion class G protein-coupled receptor with a conserved role in myelination of the peripheral nervous system. In the zebrafish, mutation of adgrg6 also results in defects in the inner ear: otic tissue fails to down-regulate versican-gene expression and morphogenesis is disrupted. We have designed a whole-animal screen that tests for rescue of both up- and down-regulated gene expression in mutant embryos, together with analysis of weak and strong alleles. From a screen of 3120 structurally diverse compounds, we have identified 68 that reduce versican-b expression in the adgrg6 mutant ear, 41 of which also restore myelin basic protein gene expression in Schwann cells of mutant embryos. Nineteen compounds unable to rescue a strong adgrg6 allele provide candidates for molecules that may interact directly with the Adgrg6 receptor. Our pipeline provides a powerful approach for identifying compounds that modulate GPCR activity, with potential impact for future drug design.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files. Source data files have been provided for Table 1 and Figure1-figure supplement 1, Figure 3, Figure 7 and Figure 7-figure supplements. Links to interactive files are given in the manuscript and in a supplemental file.

Article and author information

Author details

  1. Elvira Diamantopoulou

    Bateson Centre and Department of Biomedical Science, University of Sheffield, Sheffield, United Kingdom
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9336-7965
  2. Sarah Baxendale

    Bateson Centre and Department of Biomedical Science, University of Sheffield, Sheffield, United Kingdom
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6760-9457
  3. Antonio de la Vega de León

    Information School, University of Sheffield, Sheffield, United Kingdom
    Competing interests
    No competing interests declared.
  4. Anzar Asad

    Bateson Centre and Department of Biomedical Science, University of Sheffield, Sheffield, United Kingdom
    Competing interests
    No competing interests declared.
  5. Celia J Holdsworth

    Bateson Centre and Department of Biomedical Science, University of Sheffield, Sheffield, United Kingdom
    Competing interests
    No competing interests declared.
  6. Leila Abbas

    Bateson Centre and Department of Biomedical Science, University of Sheffield, Sheffield, United Kingdom
    Competing interests
    No competing interests declared.
  7. Valerie J Gillet

    Information School, University of Sheffield, Sheffield, United Kingdom
    Competing interests
    No competing interests declared.
  8. Giselle R Wiggin

    Sosei Heptares, Cambridge, United Kingdom
    Competing interests
    No competing interests declared.
  9. Tanya T Whitfield

    Bateson Centre and Department of Biomedical Science, University of Sheffield, Sheffield, United Kingdom
    For correspondence
    t.whitfield@sheffield.ac.uk
    Competing interests
    Tanya T Whitfield, Reviewing editor, eLife.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-1575-1504

Funding

Biotechnology and Biological Sciences Research Council (Project grant BB/J003050/1)

  • Sarah Baxendale
  • Tanya T Whitfield

University of Sheffield (PhD studentship 314420)

  • Tanya T Whitfield

Medical Research Council (G0802527)

  • Sarah Baxendale
  • Celia J Holdsworth
  • Leila Abbas
  • Tanya T Whitfield

European Union's Seventh Framework Programme (Grant agreement no. 612347)

  • Valerie J Gillet

Biotechnology and Biological Sciences Research Council (BB/R50581X/1)

  • Sarah Baxendale
  • Anzar Asad
  • Giselle R Wiggin
  • Tanya T Whitfield

Wellcome (VIP award 084551)

  • Leila Abbas
  • Tanya T Whitfield

Medical Research Council (G0700091)

  • Sarah Baxendale
  • Celia J Holdsworth
  • Leila Abbas
  • Tanya T Whitfield

Biotechnology and Biological Sciences Research Council (Project grant BB/M01021X/1)

  • Sarah Baxendale
  • Tanya T Whitfield

Biotechnology and Biological Sciences Research Council (ALERT14 equipment award BB/M012522/1)

  • Sarah Baxendale
  • Tanya T Whitfield

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: All animal work was performed under licence from the UK Home Office (P66302E4E), and approved by the University of Sheffield Ethical Review Committee (ASPA Ethical Review Process).

Reviewing Editor

  1. David A Lyons, University of Edinburgh, United Kingdom

Publication history

  1. Received: January 4, 2019
  2. Accepted: June 8, 2019
  3. Accepted Manuscript published: June 10, 2019 (version 1)
  4. Version of Record published: June 28, 2019 (version 2)

Copyright

© 2019, Diamantopoulou et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 2,044
    Page views
  • 276
    Downloads
  • 9
    Citations

Article citation count generated by polling the highest count across the following sources: Crossref, PubMed Central, Scopus.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Download citations (links to download the citations from this article in formats compatible with various reference manager tools)

Open citations (links to open the citations from this article in various online reference manager services)

Further reading

    1. Developmental Biology
    2. Neuroscience
    Baruch Haimson et al.
    Research Article Updated

    Peripheral and intraspinal feedback is required to shape and update the output of spinal networks that execute motor behavior. We report that lumbar dI2 spinal interneurons in chicks receive synaptic input from afferents and premotor neurons. These interneurons innervate contralateral premotor networks in the lumbar and brachial spinal cord, and their ascending projections innervate the cerebellum. These findings suggest that dI2 neurons function as interneurons in local lumbar circuits, are involved in lumbo-brachial coupling, and that part of them deliver peripheral and intraspinal feedback to the cerebellum. Silencing of dI2 neurons leads to destabilized stepping in posthatching day 8 hatchlings, with occasional collapses, variable step profiles, and a wide-base walking gait, suggesting that dI2 neurons may contribute to the stabilization of the bipedal gait.

    1. Developmental Biology
    2. Evolutionary Biology
    Tom Dierschke et al.
    Research Article

    Eukaryotic life cycles alternate between haploid and diploid phases and in phylogenetically diverse unicellular eukaryotes, expression of paralogous homeodomain genes in gametes primes the haploid-to-diploid transition. In the unicellular Chlorophyte alga Chlamydomonas KNOX and BELL TALE-homeodomain genes mediate this transition. We demonstrate that in the liverwort Marchantia polymorpha paternal (sperm) expression of three of five phylogenetically diverse BELL genes, MpBELL234, and maternal (egg) expression of both MpKNOX1 and MpBELL34 mediate the haploid-to-diploid transition. Loss-of-function alleles of MpKNOX1 result in zygotic arrest, whereas loss of either maternal or paternal MpBELL234 results in variable zygotic and early embryonic arrest. Expression of MpKNOX1 and MpBELL34 during diploid sporophyte development is consistent with a later role for these genes in patterning the sporophyte. These results indicate that the ancestral mechanism to activate diploid gene expression was retained in early diverging land plants and subsequently co-opted during evolution of the diploid sporophyte body.