In vivo fluorescent labeling and tracking of extracellular matrix.
Journal
Nature protocols
ISSN: 1750-2799
Titre abrégé: Nat Protoc
Pays: England
ID NLM: 101284307
Informations de publication
Date de publication:
10 2023
10 2023
Historique:
received:
18
10
2022
accepted:
02
06
2023
medline:
9
10
2023
pubmed:
10
8
2023
entrez:
9
8
2023
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 37558896
doi: 10.1038/s41596-023-00867-y
pii: 10.1038/s41596-023-00867-y
doi:
Substances chimiques
N-hydroxysuccinimide
MJE3791M4T
Succinimides
0
Extracellular Matrix Proteins
0
Types de publication
Journal Article
Review
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
2876-2890Informations de copyright
© 2023. Springer Nature Limited.
Références
Baron, C. S. & van Oudenaarden, A. Unravelling cellular relationships during development and regeneration using genetic lineage tracing. Nat. Rev. Mol. Cell Biol. 20, 753–765 (2019).
doi: 10.1038/s41580-019-0186-3
pubmed: 31690888
Hastings, J. F., Skhinas, J. N., Fey, D., Croucher, D. R. & Cox, T. R. The extracellular matrix as a key regulator of intracellular signalling networks. Br. J. Pharmacol. 176, 82–92 (2019).
doi: 10.1111/bph.14195
pubmed: 29510460
Manou, D. et al. The complex interplay between extracellular matrix and cells in tissues. Methods Mol. Biol. 1952, 1–20 (2019).
doi: 10.1007/978-1-4939-9133-4_1
pubmed: 30825161
Loganathan, R. et al. Extracellular matrix motion and early morphogenesis. Development 143, 2056–2065 (2016).
doi: 10.1242/dev.127886
pubmed: 27302396
pmcid: 4920166
Sivakumar, P. et al. New insights into extracellular matrix assembly and reorganization from dynamic imaging of extracellular matrix proteins in living osteoblasts. J. Cell Sci. 119, 1350–1360 (2006).
doi: 10.1242/jcs.02830
pubmed: 16537652
Aufschnaiter, R. et al. In vivo imaging of basement membrane movement: ECM patterning shapes Hydra polyps. J. Cell Sci. 124, 4027–4038 (2011).
doi: 10.1242/jcs.087239
pubmed: 22194305
pmcid: 3244984
Zamir, E. A., Czirók, A., Cui, C., Little, C. D. & Rongish, B. J. Mesodermal cell displacements during avian gastrulation are due to both individual cell-autonomous and convective tissue movements. Proc. Natl Acad. Sci. USA 103, 19806–19811 (2006).
doi: 10.1073/pnas.0606100103
pubmed: 17179040
pmcid: 1705812
Czirók, A., Rongish, B. J. & Little, C. D. Extracellular matrix dynamics during vertebrate axis formation. Dev. Biol. 268, 111–122 (2004).
doi: 10.1016/j.ydbio.2003.09.040
pubmed: 15031109
Dzamba, B. J. & DeSimone, D. W. Extracellular matrix (ECM) and the sculpting of embryonic tissues. Curr. Top. Dev. Biol. 130, 245–274 (2018).
doi: 10.1016/bs.ctdb.2018.03.006
pubmed: 29853179
Duffield, M. Dynamic views. In Practical App Development with Aurelia Ch. 18, 185–194 (Apress, 2018).
Pankov, R. & Yamada, K. M. Non‐radioactive quantification of fibronectin matrix assembly. Curr. Protoc. Cell Biol. 25, 1–9 (2004).
doi: 10.1002/0471143030.cb1013s25
Aper, S. J. A. et al. Colorful protein-based fluorescent probes for collagen imaging. PLoS ONE 9, 1–21 (2014).
doi: 10.1371/journal.pone.0114983
Leonard, A. K. et al. Methods for the visualization and analysis of extracellular matrix protein structure and degradation. Methods Cell Biol. 143, 79–95 (2019).
doi: 10.1016/bs.mcb.2017.08.005
Arnoldini, S. et al. Novel peptide probes to assess the tensional state of fibronectin fibers in cancer. Nat. Commun. 8, 1793 (2017).
doi: 10.1038/s41467-017-01846-0
pubmed: 29176724
pmcid: 5702617
Keeley, D. P. et al. Comprehensive endogenous tagging of basement membrane components reveals dynamic movement within the matrix scaffolding. Dev. Cell 54, 60–74 (2021).
doi: 10.1016/j.devcel.2020.05.022
Morris, J. L. et al. Live imaging of collagen deposition during skin development and repair in a collagen I–GFP fusion transgenic zebrafish line. Dev. Biol. 441, 4–11 (2018).
doi: 10.1016/j.ydbio.2018.06.001
pubmed: 29883658
pmcid: 6080847
Ohashi, T., Kiehart, D. P. & Erickson, H. P. Dual labeling of the fibronectin matrix and actin cytoskeleton with green fluorescent protein variants. J. Cell Sci. 115, 1221–1229 (2002).
doi: 10.1242/jcs.115.6.1221
pubmed: 11884521
Lu, Y. et al. Live imaging of type I collagen assembly dynamics in osteoblasts stably expressing GFP and mCherry-tagged collagen constructs. J. Bone Miner. Res. 33, 1166–1182 (2018).
doi: 10.1002/jbmr.3409
pubmed: 29461659
Poole, J. J. A. & Mostaço-guidolin, L. B. Optical microscopy and the extracellular matrix structure: a review. Cells 10, 1760 (2021).
doi: 10.3390/cells10071760
pubmed: 34359929
pmcid: 8308089
Berg, E. A. & Fishman, J. B. Labeling antibodies. Cold Spring Harb. Protoc. 2020, 252–263 (2020).
Fischer, A. et al. Neutrophils direct preexisting matrix to initiate repair in damaged tissues. Nat. Immunol. 23, 518–531 (2022).
doi: 10.1038/s41590-022-01166-6
pubmed: 35354953
pmcid: 8986538
Correa-Gallegos, D. et al. Patch repair of deep wounds by mobilized fascia. Nature 576, 287–292 (2019).
doi: 10.1038/s41586-019-1794-y
pubmed: 31776510
Wan, L. et al. Connexin43 gap junction drives fascia mobilization and repair of deep skin wounds. Matrix Biol. 97, 58–71 (2021).
doi: 10.1016/j.matbio.2021.01.005
pubmed: 33508427
Kashimoto, R. et al. Lattice-patterned collagen fibers and their dynamics in axolotl skin regeneration. iScience 25, 104524 (2022).
doi: 10.1016/j.isci.2022.104524
pubmed: 35754731
pmcid: 9213773