1. Genetics and Genomics
  2. Neuroscience

Neural mechanisms of parasite-induced summiting behavior in 'zombie' Drosophila

  1. Carolyn Elya  Is a corresponding author
  2. Danylo Lavrentovich
  3. Emily Lee
  4. Cassandra Pasadyn
  5. Jasper Duval
  6. Maya Basak
  7. Valerie Saykina
  8. Benjamin L de Bivort  Is a corresponding author
  1. Harvard University, United States
https://doi.org/10.7554/eLife.85410
Share this article
Cite this article
  1. Carolyn Elya
  2. Danylo Lavrentovich
  3. Emily Lee
  4. Cassandra Pasadyn
  5. Jasper Duval
  6. Maya Basak
  7. Valerie Saykina
  8. Benjamin L de Bivort
(2023)
Neural mechanisms of parasite-induced summiting behavior in 'zombie' Drosophila
eLife 12:e85410.
https://doi.org/10.7554/eLife.85410
  2. Download BibTeX
  3. Download .RIS

Abstract

For at least two centuries, scientists have been enthralled by the 'zombie' behaviors induced by mind-controlling parasites. Despite this interest, the mechanistic bases of these uncanny processes have remained mostly a mystery. Here, we leverage the recently established Entomophthora muscae-Drosophila melanogaster 'zombie fly' system to reveal the molecular and cellular underpinnings of summit disease, a manipulated behavior evoked by many fungal parasites. Using a new, high-throughput behavior assay to measure summiting, we discovered that summiting behavior is characterized by a burst of locomotion and requires the host circadian and neurosecretory systems, specifically DN1p circadian neurons, pars intercerebralis to corpora allata projecting (PI-CA) neurons and corpora allata (CA), who are solely responsible for juvenile hormone (JH) synthesis and release. Summiting is a fleeting phenomenon, posing a challenge for physiological and biochemical experiments requiring tissue from summiting flies. We addressed this with a machine learning classifier to identify summiting animals in real time. PI-CA neurons and CA appear to be intact in summiting animals, despite E. muscae cells invading the host brain, particularly in the superior medial protocerebrum (SMP), the neuropil that contains DN1p axons and PI-CA dendrites. The blood-brain barrier of flies late in their infection was significantly permeabilized, suggesting that factors in the hemolymph may have greater access to the central nervous system during summiting. Metabolomic analysis of hemolymph from summiting flies revealed differential abundance of several compounds compared to non-summiting flies. Transfusing the hemolymph of summiting flies into non-summiting recipients induced a burst of locomotion, demonstrating that factor(s) in the hemolymph likely cause summiting behavior. Altogether, our work reveals a neuro-mechanistic model for summiting wherein fungal cells perturb the fly's hemolymph, activating the neurohormonal pathway linking clock neurons to juvenile hormone production in the CA, ultimately inducing locomotor activity in their host.

Data availability

Data supporting these results and the analysis code are available at http://lab.debivort.org/zombie-summiting/ and https://doi.org/10.5281/zenodo.7464925. All raw behavioral tracking (centroid versus time) data are available via Harvard Dataverse at https://doi.org/10.7910/DVN/LTMCFR.

The following data sets were generated

Article and author information

Author details

  1. Carolyn Elya

    Department of Organismic and Evolutionary Biology, Harvard University, Cambridge, United States
    For correspondence
    cnelya@gmail.com
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9634-0303

  2. Danylo Lavrentovich

    Department of Organismic and Evolutionary Biology, Harvard University, Cambridge, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8432-9596

  3. Emily Lee

    Department of Organismic and Evolutionary Biology, Harvard University, Cambridge, United States
    Competing interests
    The authors declare that no competing interests exist.

  4. Cassandra Pasadyn

    Department of Organismic and Evolutionary Biology, Harvard University, Cambridge, United States
    Competing interests
    The authors declare that no competing interests exist.

  5. Jasper Duval

    Department of Organismic and Evolutionary Biology, Harvard University, Cambridge, United States
    Competing interests
    The authors declare that no competing interests exist.

  6. Maya Basak

    Department of Organismic and Evolutionary Biology, Harvard University, Cambridge, United States
    Competing interests
    The authors declare that no competing interests exist.

  7. Valerie Saykina

    Department of Organismic and Evolutionary Biology, Harvard University, Cambridge, United States
    Competing interests
    The authors declare that no competing interests exist.

  8. Benjamin L de Bivort

    Department of Organismic and Evolutionary Biology, Harvard University, Cambridge, United States
    For correspondence
    debivort@oeb.harvard.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6165-7696

Funding

Howard Hughes Medical Institute (GT11087)

  • Carolyn Elya

Harvard Quantitative Biology Initiative

  • Danylo Lavrentovich

Alfred P. Sloan Foundation (Research Fellowship)

  • Benjamin L de Bivort

Esther A. and Joseph Klingenstein Fund (Klingenstein-Simons Fellowship Award)

  • Benjamin L de Bivort

Richard and Susan Smith Family Foundation (Odyssey Award)

  • Benjamin L de Bivort

Harvard/MIT (Basic Neuroscience Grant)

  • Benjamin L de Bivort

National Science Foundation (IOS-1557913)

  • Benjamin L de Bivort

National Institute of Neurological Disorders and Stroke (1R01NS121874-01)

  • Benjamin L de Bivort

NSF-Simons Center for Mathematical and Statistical Analysis of Biology (1764269)

  • Danylo Lavrentovich

Harvard Mind Brain and Behavior Initiative (Postdoctoral Fellow Award)

  • Carolyn Elya

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

Copyright

© 2023, Elya 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

  • 4,663
    views
  • 492
    downloads
  • 20
    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. Carolyn Elya
  2. Danylo Lavrentovich
  3. Emily Lee
  4. Cassandra Pasadyn
  5. Jasper Duval
  6. Maya Basak
  7. Valerie Saykina
  8. Benjamin L de Bivort
(2023)
Neural mechanisms of parasite-induced summiting behavior in 'zombie' Drosophila
eLife 12:e85410.
https://doi.org/10.7554/eLife.85410

Share this article

https://doi.org/10.7554/eLife.85410

Categories and tags

Research organism

Further reading

    1. Genetics and Genomics

    Deterministic genetic barcoding for multiplexed behavioral and single-cell transcriptomic studies

    Jorge Blanco Mendana, Margaret Donovan ... Daryl M Gohl
    Tools and Resources

    Advances in single-cell sequencing technologies have provided novel insights into the dynamics of gene expression and cellular heterogeneity within tissues and have enabled the construction of transcriptomic cell atlases. However, linking anatomical information to transcriptomic data and positively identifying the cell types that correspond to gene expression clusters in single-cell sequencing data sets remains a challenge. We describe a straightforward genetic barcoding approach that takes advantage of the powerful genetic tools in Drosophila to allow in vivo tagging of defined cell populations. This method, called Targeted Genetically-Encoded Multiplexing (TaG-EM), involves inserting a DNA barcode just upstream of the polyadenylation site in a Gal4-inducible UAS-GFP construct so that the barcode sequence can be read out during single-cell sequencing, labeling a cell population of interest. By creating many such independently barcoded fly strains, TaG-EM enables positive identification of cell types in cell atlas projects, identification of multiplet droplets, and barcoding of experimental timepoints, conditions, and replicates. Furthermore, we demonstrate that TaG-EM barcodes can be read out using next-generation sequencing to facilitate population-scale behavioral measurements. Thus, TaG-EM has the potential to enable large-scale behavioral screens in addition to improving the ability to multiplex and reliably annotate single-cell transcriptomic experiments.

    1. Genetics and Genomics

    Intergenerational transport of double-stranded RNA in C. elegans can limit heritable epigenetic changes

    Nathan M Shugarts Devanapally, Aishwarya Sathya ... Antony M Jose
    Research Article

    RNAs in circulation carry sequence-specific regulatory information between cells in plant, animal, and host-pathogen systems. Such RNA can cross generational boundaries, as evidenced by somatic double-stranded RNA (dsRNA) in the nematode Caenorhabditis elegans silencing genes of matching sequence in progeny. Here we dissect the intergenerational path taken by dsRNA from parental circulation and discover that cytosolic import through the dsRNA importer SID-1 in the parental germline and/or developing progeny varies with developmental time and dsRNA substrates. Loss of SID-1 enhances initiation of heritable RNA silencing within the germline and causes changes in the expression of the sid-1-dependent gene sdg-1 that last for more than 100 generations after restoration of SID-1. The SDG-1 protein is enriched in perinuclear germ granules required for heritable RNA silencing but is expressed from a retrotransposon targeted by such silencing. This auto-inhibitory loop suggests how retrotransposons could persist by hosting genes that regulate their own silencing.

    1. Cell Biology
    2. Genetics and Genomics

    The PRC2.1 subcomplex opposes G1 progression through regulation of CCND1 and CCND2

    Adam D Longhurst, Kyle Wang ... David P Toczyski
    Tools and Resources

    Progression through the G1 phase of the cell cycle is the most highly regulated step in cellular division. We employed a chemogenetic approach to discover novel cellular networks that regulate cell cycle progression. This approach uncovered functional clusters of genes that altered sensitivity of cells to inhibitors of the G1/S transition. Mutation of components of the Polycomb Repressor Complex 2 rescued proliferation inhibition caused by the CDK4/6 inhibitor palbociclib, but not to inhibitors of S phase or mitosis. In addition to its core catalytic subunits, mutation of the PRC2.1 accessory protein MTF2, but not the PRC2.2 protein JARID2, rendered cells resistant to palbociclib treatment. We found that PRC2.1 (MTF2), but not PRC2.2 (JARID2), was critical for promoting H3K27me3 deposition at CpG islands genome-wide and in promoters. This included the CpG islands in the promoter of the CDK4/6 cyclins CCND1 and CCND2, and loss of MTF2 lead to upregulation of both CCND1 and CCND2. Our results demonstrate a role for PRC2.1, but not PRC2.2, in antagonizing G1 progression in a diversity of cell linages, including chronic myeloid leukemia (CML), breast cancer, and immortalized cell lines.