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

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.

Reviewing Editor

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

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).

Version 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

  • 2,694
    views
  • 335
    downloads
  • 19
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

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)

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

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

  1. Elvira Diamantopoulou
  2. Sarah Baxendale
  3. Antonio de la Vega de León
  4. Anzar Asad
  5. Celia J Holdsworth
  6. Leila Abbas
  7. Valerie J Gillet
  8. Giselle R Wiggin
  9. Tanya T Whitfield
(2019)
Identification of compounds that rescue otic and myelination defects in the zebrafish adgrg6 (gpr126) mutant
eLife 8:e44889.
https://doi.org/10.7554/eLife.44889

Share this article

https://doi.org/10.7554/eLife.44889

Further reading

    1. Developmental Biology
    2. Immunology and Inflammation
    Tobias Weinberger, Messerer Denise ... Christian Schulz
    Research Article

    Cardiac macrophages are heterogenous in phenotype and functions, which has been associated with differences in their ontogeny. Despite extensive research, our understanding of the precise role of different subsets of macrophages in ischemia/reperfusion (I/R) injury remains incomplete. We here investigated macrophage lineages and ablated tissue macrophages in homeostasis and after I/R injury in a CSF1R-dependent manner. Genomic deletion of a fms-intronic regulatory element (FIRE) in the Csf1r locus resulted in specific absence of resident homeostatic and antigen-presenting macrophages, without affecting the recruitment of monocyte-derived macrophages to the infarcted heart. Specific absence of homeostatic, monocyte-independent macrophages altered the immune cell crosstalk in response to injury and induced proinflammatory neutrophil polarization, resulting in impaired cardiac remodeling without influencing infarct size. In contrast, continuous CSF1R inhibition led to depletion of both resident and recruited macrophage populations. This augmented adverse remodeling after I/R and led to an increased infarct size and deterioration of cardiac function. In summary, resident macrophages orchestrate inflammatory responses improving cardiac remodeling, while recruited macrophages determine infarct size after I/R injury. These findings attribute distinct beneficial effects to different macrophage populations in the context of myocardial infarction.

    1. Cell Biology
    2. Developmental Biology
    Corey D Holman, Alexander P Sakers ... Patrick Seale
    Research Article

    The energy-burning capability of beige adipose tissue is a potential therapeutic tool for reducing obesity and metabolic disease, but this capacity is decreased by aging. Here, we evaluate the impact of aging on the profile and activity of adipocyte stem and progenitor cells (ASPCs) and adipocytes during the beiging process in mice. We found that aging increases the expression of Cd9 and other fibro-inflammatory genes in fibroblastic ASPCs and blocks their differentiation into beige adipocytes. Fibroblastic ASPC populations from young and aged mice were equally competent for beige differentiation in vitro, suggesting that environmental factors suppress adipogenesis in vivo. Examination of adipocytes by single nucleus RNA-sequencing identified compositional and transcriptional differences in adipocyte populations with aging and cold exposure. Notably, cold exposure induced an adipocyte population expressing high levels of de novo lipogenesis (DNL) genes, and this response was severely blunted in aged animals. We further identified Npr3, which encodes the natriuretic peptide clearance receptor, as a marker gene for a subset of white adipocytes and an aging-upregulated gene in adipocytes. In summary, this study indicates that aging blocks beige adipogenesis and dysregulates adipocyte responses to cold exposure and provides a resource for identifying cold and aging-regulated pathways in adipose tissue.