Intestinal goblet cells sample and deliver lumenal antigens by regulated endocytic uptake and transcytosis
- University of Gothenburg, Sweden
- Washington University in Saint Louis School of Medicine, United States
- University of Michigan, United States
- Washington University School of Medicine, United States
- Harvard Medical School, United States
Abstract
Intestinal goblet cells maintain the protective epithelial barrier through mucus secretion and yet sample lumenal substances for immune processing through formation of goblet cell associated antigen passages (GAPs). The cellular biology of GAPs and how these divergent processes are balanced and regulated by goblet cells remains unknown. Using high resolution light and electron microscopy, we found that in mice, GAPs were formed by an acetylcholine (ACh) dependent endocytic event remarkable for delivery of fluid phase cargo retrograde into the trans golgi network and across the cell by transcytosis - in addition to the expected transport of fluid phase cargo by endosomes to multi-vesicular bodies and lysosomes. While ACh also induced goblet cells to secrete mucins, ACh-induced GAP formation and mucin secretion were functionally independent and mediated by different receptors and signaling pathways, enabling goblet cells to differentially regulate these processes to accommodate the dynamically changing demands of the mucosal environment for barrier maintenance and sampling of lumenal substances.
Data availability
All data generated and analysed for this study are included in the manuscript and source data files for figure 8
Article and author information
Author details
Funding
National Institute of Diabetes and Digestive and Kidney Diseases (DK097317)
- Rodney D Newberry
Stiftelserna Wilhelm och Martina Lundgrens
- Jenny K Gustafsson
Åke Wiberg Stiftelse
- Jenny K Gustafsson
Jeanssons Stiftelser
- Jenny K Gustafsson
National Institute of Allergy and Infectious Diseases (AI131342)
- Rodney D Newberry
National Institute of Diabetes and Digestive and Kidney Diseases (DK109006)
- Kathryn A Knoop
National Institute of Allergy and Infectious Diseases (AI136515)
- Rodney D Newberry
National Institute of Allergy and Infectious Diseases (AI140755)
- Rodney D Newberry
National Institute of Allergy and Infectious Diseases (AI112626)
- Simon P Hogan
National Institute of Diabetes and Digestive and Kidney Diseases (DK048106)
- Wayne I Lencer
Crohn's and Colitis Foundation (34835)
- Jenny K Gustafsson
Vetenskapsrådet (2014-00366)
- Jenny K Gustafsson
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 procedures and protocols were performed in accordance with the Institutional Animal Care and Use Committee at Washington University School of Medicine (Animal Wellfare Assurance number: A-3381-01) and the Swedish animal welfare legislation and approved by the Swedish Laboratory Animal Ethical Committee in Gothenburg (Ethical permit ID number: 5.8.18-11053/2019.
Copyright
© 2021, Gustafsson 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
-
- 5,877
- views
-
- 818
- downloads
-
- 50
- 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)
Intestinal goblet cells sample and deliver lumenal antigens by regulated endocytic uptake and transcytosis
eLife 10:e67292.
https://doi.org/10.7554/eLife.67292
Further reading
-
- Cell Biology
Excessive mitochondrial fragmentation is associated with the pathologic mitochondrial dysfunction implicated in the pathogenesis of etiologically diverse diseases, including many neurodegenerative disorders. The integrated stress response (ISR) – comprising the four eIF2α kinases PERK, GCN2, PKR, and HRI – is a prominent stress-responsive signaling pathway that regulates mitochondrial morphology and function in response to diverse types of pathologic insult. This suggests that pharmacologic activation of the ISR represents a potential strategy to mitigate pathologic mitochondrial fragmentation associated with human disease. Here, we show that pharmacologic activation of the ISR kinases HRI or GCN2 promotes adaptive mitochondrial elongation and prevents mitochondrial fragmentation induced by the calcium ionophore ionomycin. Further, we show that pharmacologic activation of the ISR reduces mitochondrial fragmentation and restores basal mitochondrial morphology in patient fibroblasts expressing the pathogenic D414V variant of the pro-fusion mitochondrial GTPase MFN2 associated with neurological dysfunctions, including ataxia, optic atrophy, and sensorineural hearing loss. These results identify pharmacologic activation of ISR kinases as a potential strategy to prevent pathologic mitochondrial fragmentation induced by disease-relevant chemical and genetic insults, further motivating the pursuit of highly selective ISR kinase-activating compounds as a therapeutic strategy to mitigate mitochondrial dysfunction implicated in diverse human diseases.
-
- Cell Biology
Oscillations in insulin secretion, driven by islet Ca2+ waves, are crucial for glycemic control. Prior studies, performed with single-plane imaging, suggest that subpopulations of electrically coupled β-cells have privileged roles in leading and coordinating the propagation of Ca2+ waves. Here, we used three-dimensional (3D) light-sheet imaging to analyze the location and Ca2+ activity of single β-cells within the entire islet at >2 Hz. In contrast with single-plane studies, 3D network analysis indicates that the most highly synchronized β-cells are located at the islet center, and remain regionally but not cellularly stable between oscillations. This subpopulation, which includes ‘hub cells’, is insensitive to changes in fuel metabolism induced by glucokinase and pyruvate kinase activation. β-Cells that initiate the Ca2+ wave (leaders) are located at the islet periphery, and strikingly, change their identity over time via rotations in the wave axis. Glucokinase activation, which increased oscillation period, reinforced leader cells and stabilized the wave axis. Pyruvate kinase activation, despite increasing oscillation frequency, had no effect on leader cells, indicating the wave origin is patterned by fuel input. These findings emphasize the stochastic nature of the β-cell subpopulations that control Ca2+ oscillations and identify a role for glucokinase in spatially patterning ‘leader’ β-cells.
