eprintid: 10099402 rev_number: 14 eprint_status: archive userid: 608 dir: disk0/10/09/94/02 datestamp: 2020-06-03 13:07:01 lastmod: 2021-10-03 23:57:38 status_changed: 2020-06-03 13:07:01 type: article metadata_visibility: show creators_name: Yoshida, N creators_name: Domart, M-C creators_name: Peddie, CJ creators_name: Yakimovich, A creators_name: Mazon-Moya, MJ creators_name: Hawkins, TA creators_name: Collinson, L creators_name: Mercer, J creators_name: Frickel, E-M creators_name: Mostowy, S title: The zebrafish as a novel model for the in vivo study of Toxoplasma gondii replication and interaction with macrophages ispublished: inpress divisions: UCL divisions: B02 divisions: C08 divisions: D09 divisions: F96 keywords: CLEM, In Vivo, Macrophages, Toxoplasma gondii, Zebrafish note: © 2020 Published by The Company of Biologists Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0). abstract: Toxoplasma gondii is an obligate intracellular parasite capable of invading any nucleated cell. Three main clonal lineages (type I, II, III) exist and murine models have driven the understanding of general and strain-specific immune mechanisms underlying Toxoplasma infection. However, murine models are limited for studying parasite-leukocyte interactions in vivo, and discrepancies exist between cellular immune responses observed in mouse versus human cells. Here, we developed a zebrafish infection model to study the innate immune response to Toxoplasma in vivo By infecting the zebrafish hindbrain ventricle, and using high-resolution microscopy techniques coupled with computer vision driven automated image analysis, we reveal that Toxoplasma invades brain cells and replicates inside a parasitophorous vacuole to which type I and III parasites recruit host cell mitochondria. We also show that type II and III strains maintain a higher infectious burden than type I strains. To understand how parasites are cleared in vivo, we further analyzed Toxoplasma-macrophage interactions using time-lapse microscopy and three-dimensional correlative light and electron microscopy (3D CLEM). Time-lapse microscopy revealed that macrophages are recruited to the infection site and play a key role in Toxoplasma control. High-resolution 3D CLEM revealed parasitophorous vacuole breakage in brain cells and macrophages in vivo, suggesting that cell-intrinsic mechanisms may be used to destroy the intracellular niche of tachyzoites. Together, our results demonstrate in vivo control of Toxoplasma by macrophages, and highlight the possibility that zebrafish may be further exploited as a novel model system for discoveries within the field of parasite immunity. date: 2020-05-27 date_type: published official_url: https://doi.org/10.1242/dmm.043091 oa_status: green full_text_type: other language: eng primo: open primo_central: open_green verified: verified_manual elements_id: 1786833 doi: 10.1242/dmm.043091 pii: dmm.043091 lyricists_name: Hawkins, Thomas lyricists_id: TAHAW57 actors_name: Kalinowski, Damian actors_id: DKALI47 actors_role: owner full_text_status: public publication: Disease Models & Mechanisms event_location: England issn: 1754-8403 citation: Yoshida, N; Domart, M-C; Peddie, CJ; Yakimovich, A; Mazon-Moya, MJ; Hawkins, TA; Collinson, L; ... Mostowy, S; + view all <#> Yoshida, N; Domart, M-C; Peddie, CJ; Yakimovich, A; Mazon-Moya, MJ; Hawkins, TA; Collinson, L; Mercer, J; Frickel, E-M; Mostowy, S; - view fewer <#> (2020) The zebrafish as a novel model for the in vivo study of Toxoplasma gondii replication and interaction with macrophages. Disease Models & Mechanisms 10.1242/dmm.043091 <https://doi.org/10.1242/dmm.043091>. (In press). Green open access document_url: https://discovery.ucl.ac.uk/id/eprint/10099402/1/Hawkins_The%20zebrafish%20as%20a%20novel%20model%20for%20the%20in%20vivo%20study%20of%20Toxoplasma%20gondii%20replication%20and%20interaction%20with%20macrophages_AAM.pdf