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Fischer, A. ; Correa-Gallegos, D. ; Wannemacher, J. ; Christ, S. ; Machens, H.G.* ; Rinkevich, Y.

In vivo fluorescent labeling and tracking of extracellular matrix.

Nat. Protoc. 18, 2876-2890 (2023)
DOI PMC
Open Access Green as soon as Postprint is submitted to ZB.
Connective tissues are essential building blocks for organ development, repair and regeneration. However, we are at the early stages of understanding connective tissue dynamics. Here, we detail a method that enables in vivo fate mapping of organ extracellular matrix (ECM) by taking advantage of a crosslinking chemical reaction between amine groups and N-hydroxysuccinimide esters. This methodology enables robust labeling of ECM proteins, which complement previous affinity-based single-protein methods. This protocol is intended for entry-level scientists and the labeling step takes between 5 and 10 min. ECM 'tagging' with fluorophores using N-hydroxysuccinimide esters enables visualization of ECM spatial modifications and is particularly useful to study connective tissue dynamics in organ fibrosis, tumor stroma formation, wound healing and regeneration. This in vivo chemical fate mapping methodology is highly versatile, regardless of the tissue/organ system, and complements cellular fate-mapping techniques. Furthermore, as the basic chemistry of proteins is highly conserved between species, this method is also suitable for cross-species comparative studies of ECM dynamics.
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Publication type Article: Journal article
Document type Scientific Article
Corresponding Author
Keywords Movement; Ecm
ISSN (print) / ISBN 1754-2189
e-ISSN 1750-2799
Journal Nature Protocols
Quellenangaben Volume: 18, Issue: 10, Pages: 2876-2890 Article Number: , Supplement: ,
Publisher Nature Publishing Group
Publishing Place Heidelberger Platz 3, Berlin, 14197, Germany
Non-patent literature Publications
Reviewing status Peer reviewed
Institute(s) Institute of Regenerative Biology (IRBM)
Grants
European Foundation for the Study of Diabetes (EFSD) Anniversary Fund Program
LEO Foundation
European Research Council consolidator grant (ERC-COG)