1. Stem Cells and Regenerative Medicine

Long-term live imaging of the Drosophila adult midgut reveals real-time dynamics of division, differentiation, and loss

  1. Judy Lisette Martin  Is a corresponding author
  2. Erin Nicole Sanders
  3. Paola Moreno-Roman
  4. Leslie Ann Jaramillo Koyama
  5. Shruthi Balachandra
  6. XinXin Du
  7. Lucy Erin O'Brien  Is a corresponding author
  1. Stanford University School of Medicine, United States
https://doi.org/10.7554/eLife.36248
Share this article
Cite this article
  1. Judy Lisette Martin
  2. Erin Nicole Sanders
  3. Paola Moreno-Roman
  4. Leslie Ann Jaramillo Koyama
  5. Shruthi Balachandra
  6. XinXin Du
  7. Lucy Erin O'Brien
(2018)
Long-term live imaging of the Drosophila adult midgut reveals real-time dynamics of division, differentiation, and loss
eLife 7:e36248.
https://doi.org/10.7554/eLife.36248
  2. Download BibTeX
  3. Download .RIS

Abstract

Organ renewal is governed by the dynamics of cell division, differentiation, and loss. To study these dynamics in real time, we present a platform for extended live imaging of the adult Drosophila midgut, a premier genetic model for stem cell-based organs. A window cut into a living animal allows the midgut to be imaged while intact and physiologically functioning. This approach prolongs imaging sessions to 12-16 hours and yields movies that document cell and tissue dynamics at vivid spatiotemporal resolution. Applying a pipeline for movie processing and analysis, we uncover new, intriguing cell behaviors: that mitotic stem cells dynamically re-orient, that daughter cells use slow kinetics of Notch activation to reach a fate-specifying threshold, and that enterocytes extrude via ratcheted constriction of a junctional ring. By enabling real-time study of midgut phenomena that were previously inaccessible, our platform opens a new realm for dynamic understanding of adult organ renewal.

Data availability

All data generated or analyzed during this study are included in the manuscript and supporting files. Source data files for figures have also been uploaded to Dryad (https://dx.doi.org/10.5061/dryad.1v1g1b0).

The following data sets were generated

Article and author information

Author details

  1. Judy Lisette Martin

    Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, United States
    For correspondence
    jlmart@stanford.edu
    Competing interests
    The authors declare that no competing interests exist.

  2. Erin Nicole Sanders

    Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, United States
    Competing interests
    The authors declare that no competing interests exist.

  3. Paola Moreno-Roman

    Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, United States
    Competing interests
    The authors declare that no competing interests exist.

  4. Leslie Ann Jaramillo Koyama

    Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, United States
    Competing interests
    The authors declare that no competing interests exist.

  5. Shruthi Balachandra

    Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, United States
    Competing interests
    The authors declare that no competing interests exist.

  6. XinXin Du

    Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, United States
    Competing interests
    The authors declare that no competing interests exist.

  7. Lucy Erin O'Brien

    Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, United States
    For correspondence
    lucye@stanford.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7660-2524

Funding

National Institutes of Health (R01GM116000-01A1)

  • Judy Lisette Martin
  • Erin Nicole Sanders
  • Paola Moreno-Roman
  • Leslie Ann Jaramillo Koyama
  • Shruthi Balachandra
  • XinXin Du
  • Lucy Erin O'Brien

National Institutes of Health (1F31GM123736-01)

  • Leslie Ann Jaramillo Koyama

National Institutes of Health (Stanford Discovery Fund Innovation Program)

  • Lucy Erin O'Brien

Stanford University (Center for Biomedical Imaging at Stanford Seed Grant)

  • Judy Lisette Martin
  • Lucy Erin O'Brien

National Science Foundation (GRFP DGE-1656518)

  • Erin Nicole Sanders

National Institutes of Health (2T32GM00779038)

  • Erin Nicole Sanders
  • Leslie Ann Jaramillo Koyama

William K. Bowes, Jr. Foundation (Stanford Bio X Bowes Graduate Fellowship)

  • Paola Moreno-Roman

Stanford University (Stanford DARE (Diversifying Academia Recruiting Excellence) Fellowship)

  • Paola Moreno-Roman

National Institutes of Health (NRSA 1F32GM115065)

  • XinXin Du

Stanford University (Stanford Dean's Postdoctoral Fellowship)

  • XinXin Du

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

Reviewing Editor

  1. Allan C Spradling, Howard Hughes Medical Institute, Carnegie Institution for Science, United States

Publication history

  1. Received: February 27, 2018
  2. Accepted: November 12, 2018
  3. Accepted Manuscript published: November 14, 2018 (version 1)
  4. Version of Record published: December 3, 2018 (version 2)

Copyright

© 2018, Martin 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

  • 7,300
    Page views
  • 881
    Downloads
  • 35
    Citations

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

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. Judy Lisette Martin
  2. Erin Nicole Sanders
  3. Paola Moreno-Roman
  4. Leslie Ann Jaramillo Koyama
  5. Shruthi Balachandra
  6. XinXin Du
  7. Lucy Erin O'Brien
(2018)
Long-term live imaging of the Drosophila adult midgut reveals real-time dynamics of division, differentiation, and loss
eLife 7:e36248.
https://doi.org/10.7554/eLife.36248

Categories and tags

Research organism

Further reading

    1. Stem Cells and Regenerative Medicine

    Hepatic lipid overload triggers biliary epithelial cell activation via E2Fs

    Ece Yildiz, Gaby El Alam ... Kristina Schoonjans
    Research Article Updated

    During severe or chronic hepatic injury, biliary epithelial cells (BECs) undergo rapid activation into proliferating progenitors, a crucial step required to establish a regenerative process known as ductular reaction (DR). While DR is a hallmark of chronic liver diseases, including advanced stages of non-alcoholic fatty liver disease (NAFLD), the early events underlying BEC activation are largely unknown. Here, we demonstrate that BECs readily accumulate lipids during high-fat diet feeding in mice and upon fatty acid treatment in BEC-derived organoids. Lipid overload induces metabolic rewiring to support the conversion of adult cholangiocytes into reactive BECs. Mechanistically, we found that lipid overload activates the E2F transcription factors in BECs, which drive cell cycle progression while promoting glycolytic metabolism. These findings demonstrate that fat overload is sufficient to reprogram BECs into progenitor cells in the early stages of NAFLD and provide new insights into the mechanistic basis of this process, revealing unexpected connections between lipid metabolism, stemness, and regeneration.

    1. Stem Cells and Regenerative Medicine

    Live imaging reveals chromatin compaction transitions and dynamic transcriptional bursting during stem cell differentiation in vivo

    Dennis May, Sangwon Yun ... Valentina Greco
    Research Article Updated

    Stem cell differentiation requires dramatic changes in gene expression and global remodeling of chromatin architecture. How and when chromatin remodels relative to the transcriptional, behavioral, and morphological changes during differentiation remain unclear, particularly in an intact tissue context. Here, we develop a quantitative pipeline which leverages fluorescently-tagged histones and longitudinal imaging to track large-scale chromatin compaction changes within individual cells in a live mouse. Applying this pipeline to epidermal stem cells, we reveal that cell-to-cell chromatin compaction heterogeneity within the stem cell compartment emerges independent of cell cycle status, and instead is reflective of differentiation status. Chromatin compaction state gradually transitions over days as differentiating cells exit the stem cell compartment. Moreover, establishing live imaging of Keratin-10 (K10) nascent RNA, which marks the onset of stem cell differentiation, we find that Keratin-10 transcription is highly dynamic and largely precedes the global chromatin compaction changes associated with differentiation. Together, these analyses reveal that stem cell differentiation involves dynamic transcriptional states and gradual chromatin rearrangement.

    1. Stem Cells and Regenerative Medicine
    2. Cancer Biology

    Mesenchymal stem cell suppresses the efficacy of CAR-T toward killing lymphoma cells by modulating the microenvironment through stanniocalcin-1

    Rui Zhang, Qingxi Liu ... Wenjian Ma
    Research Article Updated

    Stem cells play critical roles both in the development of cancer and therapy resistance. Although mesenchymal stem cells (MSCs) can actively migrate to tumor sites, their impact on chimeric antigen receptor modified T cell (CAR-T) immunotherapy has been little addressed. Using an in vitro cell co-culture model including lymphoma cells and macrophages, here we report that CAR-T cell-mediated cytotoxicity was significantly inhibited in the presence of MSCs. MSCs caused an increase of CD4+ T cells and Treg cells but a decrease of CD8+ T cells. In addition, MSCs stimulated the expression of indoleamine 2,3-dioxygenase and programmed cell death-ligand 1 which contributes to the immune-suppressive function of tumors. Moreover, MSCs suppressed key components of the NLRP3 inflammasome by modulating mitochondrial reactive oxygen species release. Interestingly, all these suppressive events hindering CAR-T efficacy could be abrogated if the stanniocalcin-1 (STC1) gene, which encodes the glycoprotein hormone STC-1, was knockdown in MSC. Using xenograft mice, we confirmed that CAR-T function could also be inhibited by MSC in vivo, and STC1 played a critical role. These data revealed a novel function of MSC and STC-1 in suppressing CAR-T efficacy, which should be considered in cancer therapy and may also have potential applications in controlling the toxicity arising from the excessive immune response.