{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T14:01:36Z","timestamp":1792677696805,"version":"4.5.0"},"update-to":[{"DOI":"10.12688\/f1000research.16473.1","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2018,11,8]],"date-time":"2018-11-08T00:00:00Z","timestamp":1541635200000}}],"reference-count":58,"publisher":"F1000 Research Ltd","license":[{"start":{"date-parts":[[2018,11,8]],"date-time":"2018-11-08T00:00:00Z","timestamp":1541635200000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000364","name":"Stroke Association","doi-asserted-by":"publisher","award":["TSALECT2017\/02"],"award-info":[{"award-number":["TSALECT2017\/02"]}],"id":[{"id":"10.13039\/501100000364","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000274","name":"British Heart Foundation","doi-asserted-by":"publisher","award":["FS\/14\/64\/31291"],"award-info":[{"award-number":["FS\/14\/64\/31291"]}],"id":[{"id":"10.13039\/501100000274","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000274","name":"British Heart Foundation","doi-asserted-by":"publisher","award":["FS\/15\/67\/32038"],"award-info":[{"award-number":["FS\/15\/67\/32038"]}],"id":[{"id":"10.13039\/501100000274","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002347","name":"Bundesministerium f\u00fcr Bildung und Forschung","doi-asserted-by":"publisher","award":["MR\/M501803\/1"],"award-info":[{"award-number":["MR\/M501803\/1"]}],"id":[{"id":"10.13039\/501100002347","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000849","name":"National Centre for the Replacement, Refinement and Reduction of Animals in Research","doi-asserted-by":"publisher","award":["NC\/N002598\/1"],"award-info":[{"award-number":["NC\/N002598\/1"]}],"id":[{"id":"10.13039\/501100000849","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["f1000research.com"],"crossmark-restriction":false},"short-container-title":["F1000Res"],"abstract":"<ns5:p>Intracerebral haemorrhage (ICH) is a devastating condition with limited treatment options, and current understanding of pathophysiology is incomplete.\u00a0Spontaneous cerebral bleeding is a characteristic of the human condition that has proven difficult to recapitulate in existing pre-clinical rodent models. Zebrafish larvae are frequently used as vertebrate disease models and are associated with several advantages, including high fecundity, optical translucency and non-protected status prior to 5 days post-fertilisation. Furthermore, other groups have shown that zebrafish larvae can exhibit spontaneous ICH. The aim of this study was to investigate whether such models can be utilised to study the pathological consequences of bleeding in the brain, in the context of pre-clinical ICH research.<\/ns5:p>\n                  <ns5:p>\n                    Here, we compared existing genetic (bubblehead) and chemically inducible (atorvastatin) zebrafish larval models of spontaneous ICH and studied the subsequent disease processes. Through live, non-invasive imaging of transgenic fluorescent reporter lines and behavioural assessment we quantified brain injury, locomotor function and neuroinflammation following ICH. We show that ICH in both zebrafish larval models is comparable in timing, frequency and location. ICH results in increased brain cell death and a persistent locomotor deficit. Additionally, in haemorrhaged larvae we observed a significant increase in macrophage recruitment to the site of injury. Live\n                    <ns5:italic>in vivo<\/ns5:italic>\n                    imaging allowed us to track active macrophage-based phagocytosis of dying brain cells 24 hours after haemorrhage. Morphological analyses and quantification indicated that an increase in overall macrophage activation occurs in the haemorrhaged brain.\n                  <\/ns5:p>\n                  <ns5:p>Our study shows that in zebrafish larvae, bleeding in the brain induces quantifiable phenotypic outcomes that mimic key features of human ICH. We hope that this methodology will enable the pre-clinical ICH community to adopt the zebrafish larval model as an alternative to rodents, supporting future high throughput drug screening and as a complementary approach to elucidating crucial mechanisms associated with ICH pathophysiology.<\/ns5:p>","DOI":"10.12688\/f1000research.16473.2","type":"journal-article","created":{"date-parts":[[2018,11,8]],"date-time":"2018-11-08T11:30:10Z","timestamp":1541676610000},"page":"1617","update-policy":"https:\/\/doi.org\/10.12688\/f1000research.crossmark-policy","source":"Crossref","is-referenced-by-count":33,"title":["Using zebrafish larval models to study brain injury, locomotor and neuroinflammatory outcomes following intracerebral haemorrhage"],"prefix":"10.12688","volume":"7","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1046-9884","authenticated-orcid":false,"given":"Siobhan","family":"Crilly","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3839-5146","authenticated-orcid":false,"given":"Alexandra","family":"Njegic","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sarah E.","family":"Laurie","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elisavet","family":"Fotiou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Georgina","family":"Hudson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jack","family":"Barrington","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kirsty","family":"Webb","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Helen L.","family":"Young","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrew 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Neurol."},{"key":"ref-45","doi-asserted-by":"publisher","first-page":"791-800","DOI":"10.1007\/s13311-016-0458-y","article-title":"Clinical Trials of Immunomodulation in Ischemic Stroke.","volume":"13","author":"R Veltkamp","year":"2016","journal-title":"Neurotherapeutics."},{"key":"ref-46","doi-asserted-by":"publisher","first-page":"571-577","DOI":"10.1038\/jcbfm.2014.27","article-title":"Zebrafish models of cerebrovascular disease.","volume":"34","author":"B Walcott","year":"2014","journal-title":"J Cereb Blood Flow Metab."},{"key":"ref-47","doi-asserted-by":"publisher","first-page":"478-487","DOI":"10.1007\/s12975-016-0472-8","article-title":"Microglia Activation and Polarization After Intracerebral Hemorrhage in Mice: the Role of Protease-Activated Receptor-1.","volume":"7","author":"S Wan","year":"2016","journal-title":"Transl Stroke Res."},{"key":"ref-48","doi-asserted-by":"publisher","first-page":"432","DOI":"10.3389\/fncel.2017.00432","article-title":"Neuroprotection Exerted by Netrin-1 and Kinesin Motor KIF1A in Secondary Brain Injury following Experimental Intracerebral Hemorrhage in Rats.","volume":"11","author":"J Wang","year":"2018","journal-title":"Front Cell Neurosci."},{"key":"ref-49","article-title":"The zebrafish book: a guide for the laboratory use of zebrafish","author":"M Westerfield","year":"2000"},{"key":"ref-50","article-title":"The top 10 causes of death [Online]","year":"2017"},{"key":"ref-51","doi-asserted-by":"publisher","first-page":"53-63","DOI":"10.1016\/S1474-4422(05)70283-0","article-title":"Mechanisms of brain injury after intracerebral haemorrhage.","volume":"5","author":"G Xi","year":"2006","journal-title":"Lancet Neurol."},{"key":"ref-52","doi-asserted-by":"publisher","first-page":"152-159","DOI":"10.1016\/S1052-3057(03)00036-3","article-title":"Comparison of brain cell death and inflammatory reaction in three models of intracerebral hemorrhage in adult rats.","volume":"12","author":"M Xue","year":"2003","journal-title":"J Stroke Cerebrovasc Dis."},{"key":"ref-53","doi-asserted-by":"publisher","first-page":"1009-13","DOI":"10.1161\/STROKEAHA.114.007154","article-title":"Prognostic significance of perihematomal edema in acute intracerebral hemorrhage: pooled analysis from the intensive blood pressure reduction in acute cerebral hemorrhage trial studies.","volume":"46","author":"J Yang","year":"2015","journal-title":"Stroke."},{"key":"ref-54","doi-asserted-by":"publisher","first-page":"61","DOI":"10.3389\/fnins.2016.00061","article-title":"Interleukin-4 Ameliorates the Functional Recovery of Intracerebral Hemorrhage Through the Alternative Activation of Microglia\/Macrophage.","volume":"10","author":"J Yang","year":"2016a","journal-title":"Front Neurosci."},{"key":"ref-56","doi-asserted-by":"publisher","DOI":"10.3390\/ijms17071181","article-title":"High Morphologic Plasticity of Microglia\/Macrophages Following Experimental Intracerebral Hemorrhage in Rats.","volume":"17","author":"S Yang","year":"2016b","journal-title":"Int J Mol Sci."},{"key":"ref-55","doi-asserted-by":"publisher","first-page":"337-348","DOI":"10.1242\/dmm.027268","article-title":"Miconazole protects blood vessels from MMP9-dependent rupture and hemorrhage.","volume":"10","author":"R Yang","year":"2017","journal-title":"Dis Model Mech."},{"key":"ref-57","doi-asserted-by":"publisher","first-page":"257-266","DOI":"10.1007\/s12028-013-9861-y","article-title":"High dose deferoxamine in intracerebral hemorrhage (HI-DEF) trial: rationale, design, and methods.","volume":"19","author":"S Yeatts","year":"2013","journal-title":"Neurocrit Care."},{"key":"ref-58","doi-asserted-by":"publisher","first-page":"1033-1043","DOI":"10.1161\/STROKEAHA.116.015609","article-title":"Neuronal Death After Hemorrhagic Stroke In Vitro and In Vivo Shares Features of Ferroptosis and Necroptosis.","volume":"48","author":"M Zille","year":"2017","journal-title":"Stroke."}],"container-title":["F1000Research"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/f1000research.com\/articles\/7-1617\/v2\/pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/f1000research.com\/articles\/7-1617\/v2\/xml","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/f1000research.com\/articles\/7-1617\/v2\/iparadigms","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2020,9,29]],"date-time":"2020-09-29T04:00:26Z","timestamp":1601352026000},"score":1,"resource":{"primary":{"URL":"https:\/\/f1000research.com\/articles\/7-1617\/v2"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,11,8]]},"references-count":58,"URL":"https:\/\/doi.org\/10.12688\/f1000research.16473.2","relation":{"has-review":[{"id-type":"doi","id":"10.5256\/f1000research.18516.r40436","asserted-by":"subject"},{"id-type":"doi","id":"10.5256\/f1000research.18516.r40436","asserted-by":"object"}]},"ISSN":["2046-1402"],"issn-type":[{"value":"2046-1402","type":"electronic"}],"subject":[],"published":{"date-parts":[[2018,11,8]]},"assertion":[{"value":"Indexed","URL":"https:\/\/f1000research.com\/articles\/7-1617\/v2#article-reports","order":0,"name":"referee-status","label":"Referee status","group":{"name":"current-referee-status","label":"Current Referee Status"}},{"value":"10.5256\/f1000research.18006.r39725, Claire L. Gibson, University of Nottingham , Nottingham, UK, 24 Oct 2018, version 1, 1 approved, 1 approved with reservations","URL":"https:\/\/f1000research.com\/articles\/7-1617\/v1#referee-response-39725","order":0,"name":"referee-response-39725","label":"Referee Report","group":{"name":"article-reports","label":"Article Reports"}},{"value":"<b>siobhan crilly<\/b>; \n<i>Posted: 02 Nov 2018<\/i>; Dear Prof Gibson,Many thanks indeed for reviewing our manuscript and your helpful comments. Please see below for responses to your specific questions.\n<b><i>1. Authors state that a proportion of ATV-treated embryos did not develop ICH and therefore were used as controls \u2013 what proportion was unsuccessful?&nbsp; Can the authors suggest why not all were successful? Did this result in a group of animals forming a control group which was treated separately to another control group (i.e. the untreated siblings)?<\/i><\/b>The ATV treated non-haemorrhaged group formed the ICH- control group for the ATV model. As you suggest, this group was separate from the untreated control group, and our subsequent analyses compared all 3 groups (i.e. untreated, ICH- and ICH+ groups), as presented in Figure 2B and 3C. The proportion of ICH- controls varied with ATV dose, as presented in Figure 1B. For our cell death and motility assays, we used an ATV dosage of 1.0&micro;g\/ml, which resulted in approximately 20% of non-haemorrhaged animals, which we used as our ICH- group. We currently cannot explain why a proportion of ATV-treated larvae do not develop ICH. However as zebrafish are a relatively outbred model species, we hypothesise that a genetic component may explain increased neuroendothelial stability in some animals.\n<b><i>2. Not clear to me (although I am a rodent researcher!) how a brief exposure to anaesthesia at 72hpf would remove locomotor function bias or was necessary? Couldn\u2019t a baseline measurement be taken and data compared to this? The authors states that the exposure to this brief anaesthesia was at 72hpf but also state functional analyses were done at 72, 96 and 120 hpf \u2013 did the first time point occur coincidentally with the brief anaesthesia?<\/i><\/b>Following fertilisation, we routinely plate n=50-100 embryos per petri dish. At 72hpf, natural variation in swimming behaviour\/ability exists between individual larvae. For allocation to the locomotion assay, larvae are transferred from the petri dish into a 24 well plate using a pipette. To avoid always \u2018catching\u2019 the weakest\/slowest swimmers, we briefly anaesthetise the animals so that larvae are randomly allocated to the assay. These fish were segregated at 72 hpf for repeat assays at 96 and 120 hpf and so there was no need to re-anaesthetise the animals for these later timepoints.Kind regardsSiobhan Crilly &amp; Paul Kasher","URL":"https:\/\/f1000research.com\/articles\/7-1617\/v2#referee-comment-4102","order":1,"name":"referee-comment-4102","label":"Referee Comment","group":{"name":"article-reports","label":"Article Reports"}},{"value":"10.5256\/f1000research.18516.r40436, Marietta Zille, University of L\u00fcbeck, Institute for Experimental and Clinical Pharmacology and Toxicology, L\u00fcbeck, Germany, 12 Nov 2018, version 2, indexed","URL":"https:\/\/f1000research.com\/articles\/7-1617\/v2#referee-response-40436","order":2,"name":"referee-response-40436","label":"Referee Report","group":{"name":"article-reports","label":"Article Reports"}},{"value":"This study was supported by the NC3Rs (NC\/N002598\/1), Stroke Association (TSA LECT 2017\/02), ERA-NET NEURON (MR\/M501803\/1) and The British Heart Foundation (FS\/14\/64\/31291, FS\/15\/67\/32038). We are also particularly thankful to The Natalie Kate Moss Trust for their continued financial support.","order":3,"name":"grant-information","label":"Grant Information"},{"value":"This is an open access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.Data associated with the article are available under the terms of the Creative Commons Zero \"No rights reserved\" data waiver (CC0 1.0 Public domain dedication).","order":0,"name":"copyright-info","label":"Copyright"}]}}