Using Zebrafish to Dissect the Interaction of Mycobacteria with the Autophagic Machinery in Macrophages
Existing drug treatment against tuberculosis is no match against the increasing number of multi-drug resistant strains of its causative agent, <i>Mycobacterium tuberculosis</i> (<i>Mtb</i>). A better understanding of how mycobacteria subvert the host immune defenses is crucia...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-06-01
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| Series: | Biology |
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| Online Access: | https://www.mdpi.com/2079-7737/12/6/817 |
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| author | Salomé Muñoz-Sánchez Mónica Varela Michiel van der Vaart Annemarie H. Meijer |
| author_facet | Salomé Muñoz-Sánchez Mónica Varela Michiel van der Vaart Annemarie H. Meijer |
| author_sort | Salomé Muñoz-Sánchez |
| collection | DOAJ |
| description | Existing drug treatment against tuberculosis is no match against the increasing number of multi-drug resistant strains of its causative agent, <i>Mycobacterium tuberculosis</i> (<i>Mtb</i>). A better understanding of how mycobacteria subvert the host immune defenses is crucial for developing novel therapeutic strategies. A potential approach is enhancing the activity of the autophagy machinery, which can direct bacteria to autophagolysosomal degradation. However, the interplay specifics between mycobacteria and the autophagy machinery must be better understood. Here, we analyzed live imaging data from the zebrafish model of tuberculosis to characterize mycobacteria-autophagy interactions during the early stages of infection in vivo. For high-resolution imaging, we microinjected fluorescent <i>Mycobacterium marinum</i> (<i>Mm</i>) into the tail fin tissue of zebrafish larvae carrying the GFP-LC3 autophagy reporter. We detected phagocytosed <i>Mm</i> clusters and LC3-positive <i>Mm</i>-containing vesicles within the first hour of infection. LC3 associations with these vesicles were transient and heterogeneous, ranging from simple vesicles to complex compound structures, dynamically changing shape by fusions between <i>Mm</i>-containing and empty vesicles. LC3-<i>Mm</i>-vesicles could adopt elongated shapes during cell migration or alternate between spacious and compact morphologies. LC3-<i>Mm</i>-vesicles were also observed in cells reverse migrating from the infection site, indicating that the autophagy machinery fails to control infection before tissue dissemination. |
| first_indexed | 2024-03-11T02:44:45Z |
| format | Article |
| id | doaj.art-6630e805fb06483e896d695771dd8e07 |
| institution | Directory Open Access Journal |
| issn | 2079-7737 |
| language | English |
| last_indexed | 2024-03-11T02:44:45Z |
| publishDate | 2023-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biology |
| spelling | doaj.art-6630e805fb06483e896d695771dd8e072023-11-18T09:22:58ZengMDPI AGBiology2079-77372023-06-0112681710.3390/biology12060817Using Zebrafish to Dissect the Interaction of Mycobacteria with the Autophagic Machinery in MacrophagesSalomé Muñoz-Sánchez0Mónica Varela1Michiel van der Vaart2Annemarie H. Meijer3Institute of Biology Leiden, Leiden University, Einsteinweg 55, 2333 CC Leiden, The NetherlandsInstitute of Biology Leiden, Leiden University, Einsteinweg 55, 2333 CC Leiden, The NetherlandsInstitute of Biology Leiden, Leiden University, Einsteinweg 55, 2333 CC Leiden, The NetherlandsInstitute of Biology Leiden, Leiden University, Einsteinweg 55, 2333 CC Leiden, The NetherlandsExisting drug treatment against tuberculosis is no match against the increasing number of multi-drug resistant strains of its causative agent, <i>Mycobacterium tuberculosis</i> (<i>Mtb</i>). A better understanding of how mycobacteria subvert the host immune defenses is crucial for developing novel therapeutic strategies. A potential approach is enhancing the activity of the autophagy machinery, which can direct bacteria to autophagolysosomal degradation. However, the interplay specifics between mycobacteria and the autophagy machinery must be better understood. Here, we analyzed live imaging data from the zebrafish model of tuberculosis to characterize mycobacteria-autophagy interactions during the early stages of infection in vivo. For high-resolution imaging, we microinjected fluorescent <i>Mycobacterium marinum</i> (<i>Mm</i>) into the tail fin tissue of zebrafish larvae carrying the GFP-LC3 autophagy reporter. We detected phagocytosed <i>Mm</i> clusters and LC3-positive <i>Mm</i>-containing vesicles within the first hour of infection. LC3 associations with these vesicles were transient and heterogeneous, ranging from simple vesicles to complex compound structures, dynamically changing shape by fusions between <i>Mm</i>-containing and empty vesicles. LC3-<i>Mm</i>-vesicles could adopt elongated shapes during cell migration or alternate between spacious and compact morphologies. LC3-<i>Mm</i>-vesicles were also observed in cells reverse migrating from the infection site, indicating that the autophagy machinery fails to control infection before tissue dissemination.https://www.mdpi.com/2079-7737/12/6/817zebrafishmacrophageslive imaginginfectioninnate immunityautophagy |
| spellingShingle | Salomé Muñoz-Sánchez Mónica Varela Michiel van der Vaart Annemarie H. Meijer Using Zebrafish to Dissect the Interaction of Mycobacteria with the Autophagic Machinery in Macrophages Biology zebrafish macrophages live imaging infection innate immunity autophagy |
| title | Using Zebrafish to Dissect the Interaction of Mycobacteria with the Autophagic Machinery in Macrophages |
| title_full | Using Zebrafish to Dissect the Interaction of Mycobacteria with the Autophagic Machinery in Macrophages |
| title_fullStr | Using Zebrafish to Dissect the Interaction of Mycobacteria with the Autophagic Machinery in Macrophages |
| title_full_unstemmed | Using Zebrafish to Dissect the Interaction of Mycobacteria with the Autophagic Machinery in Macrophages |
| title_short | Using Zebrafish to Dissect the Interaction of Mycobacteria with the Autophagic Machinery in Macrophages |
| title_sort | using zebrafish to dissect the interaction of mycobacteria with the autophagic machinery in macrophages |
| topic | zebrafish macrophages live imaging infection innate immunity autophagy |
| url | https://www.mdpi.com/2079-7737/12/6/817 |
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