Gain-of-function mutations in the UNC-2/CaV2α channel lead to excitation-dominant synaptic transmission in C. elegans

  1. Yung-Chi Huang
  2. Jennifer K Pirri
  3. Diego Rayes
  4. Shangbang Gao
  5. Ben Mulcahy
  6. Jeff Grant
  7. Yasunori Saheki
  8. Michael M Francis
  9. Mei Zhen
  10. Mark J Alkema  Is a corresponding author
  1. University of Massachusetts Medical School, United States
  2. Mount Sinai Hospital, Canada
  3. The Rockefeller University, United States

Abstract

Mutations in pre-synaptic voltage gated calcium channels can lead to familial hemiplegic migraine type 1 (FHM1). While mammalian studies indicate that the migraine brain is hyperexcitable due to enhanced excitation or reduced inhibition, the molecular and cellular mechanisms underlying this excitatory/inhibitory (E/I) imbalance are poorly understood. We identified a gain-of-function (gf) mutation in the Caenorhabditis elegans CaV2 channel α1 subunit, UNC-2, which leads to increased calcium currents. unc-2(zf35gf) mutants exhibit hyperactivity and seizure-like motor behaviors. Expression of the unc-2 gene with FHM1 substitutions R192Q and S218L leads to hyperactivity similar to that of unc-2(zf35gf) mutants. unc-2(zf35gf) mutants display increased cholinergic- and decreased GABAergic-transmission. Moreover, increased cholinergic transmission in unc-2(zf35gf) mutants leads to an increase of cholinergic synapses and a TAX-6/calcineurin dependent reduction of GABA synapses. Our studies reveal mechanisms through which CaV2 gain-of-function mutations disrupt excitation-inhibition balance in the nervous system.

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 all figures.

Article and author information

Author details

  1. Yung-Chi Huang

    Department of Neurobiology, University of Massachusetts Medical School, Worcester, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Jennifer K Pirri

    Department of Neurobiology, University of Massachusetts Medical School, Worcester, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Diego Rayes

    Department of Neurobiology, University of Massachusetts Medical School, Worcester, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Shangbang Gao

    Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, Canada
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5431-4628
  5. Ben Mulcahy

    Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, Canada
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-3336-245X
  6. Jeff Grant

    Department of Neurobiology, University of Massachusetts Medical School, Worcester, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Yasunori Saheki

    Lulu and Anthony Wang Laboratory of Neural Circuits and Behavior, The Rockefeller University, New York, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. Michael M Francis

    Department of Neurobiology, University of Massachusetts Medical School, Worcester, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8076-6668
  9. Mei Zhen

    Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, Canada
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-0086-9622
  10. Mark J Alkema

    Department of Neurobiology, University of Massachusetts Medical School, Worcester, United States
    For correspondence
    mark.alkema@umassmed.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-1311-5179

Funding

National Institutes of Health (GM084491)

  • Mark J Alkema

National Institutes of Health (NS082525)

  • Mei Zhen
  • Mark J Alkema

Canadian Institutes of Health Research (154274)

  • Mei Zhen

Natural Sciences and Engineering Research Council of Canada (RGPIN-2017-06738)

  • Mei Zhen

National Natural Science Foundation of China (31671052)

  • Shangbang Gao

National Institutes of Health (NS064263)

  • Michael M Francis

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

Reviewing Editor

  1. Piali Sengupta, Brandeis University, United States

Publication history

  1. Received: February 18, 2019
  2. Accepted: July 30, 2019
  3. Accepted Manuscript published: July 31, 2019 (version 1)
  4. Accepted Manuscript updated: August 5, 2019 (version 2)
  5. Version of Record published: August 28, 2019 (version 3)

Copyright

© 2019, Huang 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

  • 2,651
    Page views
  • 339
    Downloads
  • 8
    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)

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. Yung-Chi Huang
  2. Jennifer K Pirri
  3. Diego Rayes
  4. Shangbang Gao
  5. Ben Mulcahy
  6. Jeff Grant
  7. Yasunori Saheki
  8. Michael M Francis
  9. Mei Zhen
  10. Mark J Alkema
(2019)
Gain-of-function mutations in the UNC-2/CaV2α channel lead to excitation-dominant synaptic transmission in C. elegans
eLife 8:e45905.
https://doi.org/10.7554/eLife.45905

Further reading

    1. Cell Biology
    2. Genetics and Genomics
    Marlene Henríquez-Urrutia et al.
    Research Article

    Circadian clocks are important for an individual’s fitness, and recent studies have underlined their role in the outcome of biological interactions. However, the relevance of circadian clocks in fungal-fungal interactions remains largely unexplored. We sought to characterize a functional clock in the biocontrol agent Trichoderma atroviride to assess its importance in the mycoparasitic interaction against the phytopathogen Botrytis cinerea. By utilizing luciferase reporters to monitor the T. atroviride core-clock, we confirmed the existence of circadian oscillations of ~26h that are temperature-compensated and modulated by environmental cues such as light and temperature. Notably, the presence of such rhythms appears to be highly dependent on the nutritional composition of the media. Heterologous expression of the T. atroviride negative clock component (tafrq) in a clock null (Δfrq) strain of Neurospora crassa restored core clock function in the latter fungus, with the same period observed in T. atroviride, confirming the role of tafrq as a bona fide core-clock component. Confrontation assays between wild-type and clock mutant strains of T. atroviride and B. cinerea, in constant light or darkness, revealed an inhibitory effect of light on T. atroviride's mycoparasitic capabilities. Interestingly, when confrontation assays were performed under light/dark cycles, T. atroviride's overgrowth capacity was enhanced when inoculations were at dawn compared to dusk. Deleting the core-clock negative element FRQ in B. cinerea, but not in T. atroviride, was vital for the daily differential phenotype, suggesting that the B. cinerea clock has a more significant influence on the result of this interaction. Additionally, we observed that T. atroviride clock components modulate development and secondary metabolism in this fungus, affecting the production of several molecules, including volatile compounds, such as 6-pentyl-α-pyrone (6-PP). Notably, we detected the rhythmic production of distinct T. atroviride volatile organic compounds (VOCs), which depended on its circadian clock. Thus, this study provides evidence on how clock components impact diverse aspects of T. atroviride lifestyle and how daily changes modulate fungal interactions and dynamics.

    1. Cancer Biology
    2. Genetics and Genomics
    Gabriel Renaud et al.
    Research Article Updated

    Sequencing of cell-free DNA (cfDNA) is currently being used to detect cancer by searching both for mutational and non-mutational alterations. Recent work has shown that the length distribution of cfDNA fragments from a cancer patient can inform tumor load and type. Here, we propose non-negative matrix factorization (NMF) of fragment length distributions as a novel and completely unsupervised method for studying fragment length patterns in cfDNA. Using shallow whole-genome sequencing (sWGS) of cfDNA from a cohort of patients with metastatic castration-resistant prostate cancer (mCRPC), we demonstrate how NMF accurately infers the true tumor fragment length distribution as an NMF component - and that the sample weights of this component correlate with ctDNA levels (r=0.75). We further demonstrate how using several NMF components enables accurate cancer detection on data from various early stage cancers (AUC = 0.96). Finally, we show that NMF, when applied across genomic regions, can be used to discover fragment length signatures associated with open chromatin.