Intravital imaging of metastasis in adult Zebrafish
Background Metastasis is a major clinical problem whose biology is not yet fully understood. This lack of understanding is especially true for the events at the metastatic site, which include arrest, extravasation, and growth into macrometastases. Intravital imaging is a powerful te...
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| Format: | Article |
| Language: | English |
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BioMed Central
2017
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| Online Access: | http://hdl.handle.net/1721.1/111672 https://orcid.org/0000-0001-5215-6777 https://orcid.org/0000-0001-7603-8396 |
| _version_ | 1811086961782292480 |
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| author | Benjamin, David Colin Hynes, Richard O. |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Benjamin, David Colin Hynes, Richard O. |
| author_sort | Benjamin, David Colin |
| collection | MIT |
| description | Background
Metastasis is a major clinical problem whose biology is not yet fully understood. This lack of understanding is especially true for the events at the metastatic site, which include arrest, extravasation, and growth into macrometastases. Intravital imaging is a powerful technique that has shown great promise in increasing our understanding of these events. To date, most intravital imaging studies have been performed in mice, which has limited its adoption. Zebrafish are also a common system for the intravital imaging of metastasis. However, as imaging in embryos is technically simpler, relatively few studies have used adult zebrafish to study metastasis and none have followed individual cells at the metastatic site over time. The aim of this study was to demonstrate that adult casper zebrafish offer a convenient model system for performing intravital imaging of the metastatic site over time with single-cell resolution.
Methods
ZMEL1 zebrafish melanoma cells were injected into 6 to 10-week-old casper fish using an intravenous injection protocol. Because casper fish are transparent even as adults, they could be imaged without surgical intervention. Individual cells were followed over the course of 2 weeks as they arrested, extravasated, and formed macroscopic metastases. Results
Our injection method reliably delivered cells into circulation and led to the formation of tumors in multiple organs. Cells in the skin and sub-dermal muscle could be imaged at high resolution over 2 weeks using confocal microscopy. Arrest was visualized and determined to be primarily due to size restriction. Following arrest, extravasation was seen to occur between 1 and 6 days post-injection. Once outside of the vasculature, cells were observed migrating as well as forming protrusions.
Conclusions
Casper fish are a useful model for studying the events at the metastatic site using intravital imaging. The protocols described in this study are relatively simple. Combined with the reasonably low cost of zebrafish, they offer to increase access to intravital imaging. |
| first_indexed | 2024-09-23T13:37:34Z |
| format | Article |
| id | mit-1721.1/111672 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T13:37:34Z |
| publishDate | 2017 |
| publisher | BioMed Central |
| record_format | dspace |
| spelling | mit-1721.1/1116722022-10-01T16:08:40Z Intravital imaging of metastasis in adult Zebrafish Benjamin, David Colin Hynes, Richard O. Massachusetts Institute of Technology. Department of Biology Koch Institute for Integrative Cancer Research at MIT Benjamin, David Colin Hynes, Richard O. Background Metastasis is a major clinical problem whose biology is not yet fully understood. This lack of understanding is especially true for the events at the metastatic site, which include arrest, extravasation, and growth into macrometastases. Intravital imaging is a powerful technique that has shown great promise in increasing our understanding of these events. To date, most intravital imaging studies have been performed in mice, which has limited its adoption. Zebrafish are also a common system for the intravital imaging of metastasis. However, as imaging in embryos is technically simpler, relatively few studies have used adult zebrafish to study metastasis and none have followed individual cells at the metastatic site over time. The aim of this study was to demonstrate that adult casper zebrafish offer a convenient model system for performing intravital imaging of the metastatic site over time with single-cell resolution. Methods ZMEL1 zebrafish melanoma cells were injected into 6 to 10-week-old casper fish using an intravenous injection protocol. Because casper fish are transparent even as adults, they could be imaged without surgical intervention. Individual cells were followed over the course of 2 weeks as they arrested, extravasated, and formed macroscopic metastases. Results Our injection method reliably delivered cells into circulation and led to the formation of tumors in multiple organs. Cells in the skin and sub-dermal muscle could be imaged at high resolution over 2 weeks using confocal microscopy. Arrest was visualized and determined to be primarily due to size restriction. Following arrest, extravasation was seen to occur between 1 and 6 days post-injection. Once outside of the vasculature, cells were observed migrating as well as forming protrusions. Conclusions Casper fish are a useful model for studying the events at the metastatic site using intravital imaging. The protocols described in this study are relatively simple. Combined with the reasonably low cost of zebrafish, they offer to increase access to intravital imaging. National Cancer Institute (U.S.) (Grant P30-CA14051) National Institutes of Health (U.S.) (Grant 5-T32-GM007287) 2017-10-02T18:06:56Z 2017-10-02T18:06:56Z 2017-09 2017-04 2017-10-01T03:51:56Z Article http://purl.org/eprint/type/JournalArticle 1471-2407 http://hdl.handle.net/1721.1/111672 Benjamin, David C. and Hynes, Richard O. "Intravital imaging of metastasis in adult Zebrafish." BMC Cancer 17 (September 2017): 660 © 2017 The Author(s) https://orcid.org/0000-0001-5215-6777 https://orcid.org/0000-0001-7603-8396 en http://dx.doi.org/10.1186/s12885-017-3647-0 BMC Cancer Creative Commons Attribution http://creativecommons.org/licenses/by/4.0/ The Author(s). application/pdf BioMed Central BioMed Central |
| spellingShingle | Benjamin, David Colin Hynes, Richard O. Intravital imaging of metastasis in adult Zebrafish |
| title | Intravital imaging of metastasis in adult Zebrafish |
| title_full | Intravital imaging of metastasis in adult Zebrafish |
| title_fullStr | Intravital imaging of metastasis in adult Zebrafish |
| title_full_unstemmed | Intravital imaging of metastasis in adult Zebrafish |
| title_short | Intravital imaging of metastasis in adult Zebrafish |
| title_sort | intravital imaging of metastasis in adult zebrafish |
| url | http://hdl.handle.net/1721.1/111672 https://orcid.org/0000-0001-5215-6777 https://orcid.org/0000-0001-7603-8396 |
| work_keys_str_mv | AT benjamindavidcolin intravitalimagingofmetastasisinadultzebrafish AT hynesrichardo intravitalimagingofmetastasisinadultzebrafish |