Whole-body cellular mapping in mouse using standard IgG antibodies.
Journal
Nature biotechnology
ISSN: 1546-1696
Titre abrégé: Nat Biotechnol
Pays: United States
ID NLM: 9604648
Informations de publication
Date de publication:
10 Jul 2023
10 Jul 2023
Historique:
received:
12
08
2022
accepted:
26
05
2023
medline:
11
7
2023
pubmed:
11
7
2023
entrez:
10
7
2023
Statut:
aheadofprint
Résumé
Whole-body imaging techniques play a vital role in exploring the interplay of physiological systems in maintaining health and driving disease. We introduce wildDISCO, a new approach for whole-body immunolabeling, optical clearing and imaging in mice, circumventing the need for transgenic reporter animals or nanobody labeling and so overcoming existing technical limitations. We identified heptakis(2,6-di-O-methyl)-β-cyclodextrin as a potent enhancer of cholesterol extraction and membrane permeabilization, enabling deep, homogeneous penetration of standard antibodies without aggregation. WildDISCO facilitates imaging of peripheral nervous systems, lymphatic vessels and immune cells in whole mice at cellular resolution by labeling diverse endogenous proteins. Additionally, we examined rare proliferating cells and the effects of biological perturbations, as demonstrated in germ-free mice. We applied wildDISCO to map tertiary lymphoid structures in the context of breast cancer, considering both primary tumor and metastases throughout the mouse body. An atlas of high-resolution images showcasing mouse nervous, lymphatic and vascular systems is accessible at http://discotechnologies.org/wildDISCO/atlas/index.php .
Identifiants
pubmed: 37430076
doi: 10.1038/s41587-023-01846-0
pii: 10.1038/s41587-023-01846-0
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : Deutsche Forschungsgemeinschaft (German Research Foundation)
ID : EXC 2145 SyNergy, ID 390857198
Organisme : Deutsche Forschungsgemeinschaft (German Research Foundation)
ID : SFB 1052, project A9; TR 296 project 03
Organisme : Bundesministerium für Bildung und Forschung (Federal Ministry of Education and Research)
ID : 01KX2121
Informations de copyright
© 2023. The Author(s).
Références
Richardson, D. S. et al. Tissue clearing. Nat. Rev. Meth. Primers 1, 85 (2021).
doi: 10.1038/s43586-021-00080-9
Ueda, H. R. et al. Tissue clearing and its applications in neuroscience. Nat. Rev. Neurosci. 21, 61–79 (2020).
doi: 10.1038/s41583-019-0250-1
pubmed: 31896771
pmcid: 8121164
Chung, K. et al. Structural and molecular interrogation of intact biological systems. Nature 497, 332–337 (2013).
doi: 10.1038/nature12107
pubmed: 23575631
pmcid: 4092167
Livet, J. et al. Transgenic strategies for combinatorial expression of fluorescent proteins in the nervous system. Nature 450, 56–62 (2007).
doi: 10.1038/nature06293
pubmed: 17972876
Cai, R. et al. Panoptic imaging of transparent mice reveals whole-body neuronal projections and skull–meninges connections. Nat. Neurosci. 22, 317–327 (2019).
doi: 10.1038/s41593-018-0301-3
pubmed: 30598527
Rios, A. C. et al. Intraclonal plasticity in mammary tumors revealed through large-scale single-cell resolution 3D imaging. Cancer Cell 35, 618–632.e616 (2019).
doi: 10.1016/j.ccell.2019.02.010
pubmed: 30930118
Yang, B. et al. Single-cell phenotyping within transparent intact tissue through whole-body clearing. Cell 158, 945–958 (2014).
doi: 10.1016/j.cell.2014.07.017
pubmed: 25088144
pmcid: 4153367
Park, Y.-G. et al. Protection of tissue physicochemical properties using polyfunctional crosslinkers. Nat. Biotechnol. 37, 73–83 (2019).
doi: 10.1038/nbt.4281
Ku, T. et al. Elasticizing tissues for reversible shape transformation and accelerated molecular labeling. Nat. Methods 17, 609–613 (2020).
doi: 10.1038/s41592-020-0823-y
pubmed: 32424271
pmcid: 8056749
Murray, E. et al. Simple, scalable proteomic imaging for high-dimensional profiling of intact systems. Cell 163, 1500–1514 (2015).
doi: 10.1016/j.cell.2015.11.025
pubmed: 26638076
pmcid: 5275966
Renier, N. et al. iDISCO: a simple, rapid method to immunolabel large tissue samples for volume imaging. Cell 159, 896–910 (2014).
doi: 10.1016/j.cell.2014.10.010
pubmed: 25417164
Ertürk, A. et al. Three-dimensional imaging of solvent-cleared organs using 3DISCO. Nat. Protoc. 7, 1983–1995 (2012).
doi: 10.1038/nprot.2012.119
pubmed: 23060243
Susaki, E. A. et al. Whole-brain imaging with single-cell resolution using chemical cocktails and computational analysis. Cell 157, 726–739 (2014).
doi: 10.1016/j.cell.2014.03.042
pubmed: 24746791
Dodt, H.-U. et al. Ultramicroscopy: three-dimensional visualization of neuronal networks in the whole mouse brain. Nat. Methods 4, 331–336 (2007).
doi: 10.1038/nmeth1036
pubmed: 17384643
Nudell, V. et al. HYBRiD: hydrogel-reinforced DISCO for clearing mammalian bodies. Nat. Methods 19, 479–485 (2022).
doi: 10.1038/s41592-022-01427-0
pubmed: 35347322
pmcid: 9337799
Zhao, S. et al. Cellular and molecular probing of intact human organs. Cell 180, 796–812.e719 (2020).
doi: 10.1016/j.cell.2020.01.030
pubmed: 32059778
pmcid: 7557154
Belle, M. et al. Tridimensional visualization and analysis of early human development. Cell 169, 161–173.e112 (2017).
doi: 10.1016/j.cell.2017.03.008
pubmed: 28340341
Feng, G. et al. Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP. Neuron 28, 41–51 (2000).
doi: 10.1016/S0896-6273(00)00084-2
pubmed: 11086982
Mahammad, S. & Parmryd, I. in Methods in Membrane Lipids (ed. Owen, D. M.) 91–102 (Springer, 2015).
Serno, T., Geidobler, R. & Winter, G. Protein stabilization by cyclodextrins in the liquid and dried state. Adv. Drug Delivery Rev. 63, 1086–1106 (2011).
doi: 10.1016/j.addr.2011.08.003
Bernier-Latmani, J. & Petrova, T. V. High-resolution 3D analysis of mouse small-intestinal stroma. Nat. Protoc. 11, 1617–1629 (2016).
doi: 10.1038/nprot.2016.092
pubmed: 27560169
Cugurra, A. et al. Skull and vertebral bone marrow are myeloid cell reservoirs for the meninges and CNS parenchyma. Science 373, eabf7844 (2021).
doi: 10.1126/science.abf7844
pubmed: 34083447
pmcid: 8863069
Louveau, A. et al. Structural and functional features of central nervous system lymphatic vessels. Nature 523, 337–341 (2015).
doi: 10.1038/nature14432
pubmed: 26030524
pmcid: 4506234
Jacob, L. et al. Conserved meningeal lymphatic drainage circuits in mice and humans. J. Exp. Med. 219, e20220035 (2022).
doi: 10.1084/jem.20220035
pubmed: 35776089
pmcid: 9253621
Luczynski, P. et al. Growing up in a bubble: using germ-free animals to assess the influence of the gut microbiota on brain and behavior. Int. J. Neuropsychopharmacolog. 19, pyw020 (2016).
doi: 10.1093/ijnp/pyw020
Desbonnet, L., Clarke, G., Shanahan, F., Dinan, T. G. & Cryan, J. F. Microbiota is essential for social development in the mouse. Mol. Psychiatry 19, 146–148 (2014).
doi: 10.1038/mp.2013.65
pubmed: 23689536
Heijtz, R. D. et al. Normal gut microbiota modulates brain development and behavior. Proc. Natl Acad. Sci. USA 108, 3047–3052 (2011).
doi: 10.1073/pnas.1010529108
pmcid: 3041077
Fülling, C., Dinan, T. G. & Cryan, J. F. Gut microbe to brain signaling: what happens in vagus…. Neuron 101, 998–1002 (2019).
doi: 10.1016/j.neuron.2019.02.008
pubmed: 30897366
Schumacher, T. N. & Thommen, D. S. Tertiary lymphoid structures in cancer. Science 375, eabf9419 (2022).
doi: 10.1126/science.abf9419
pubmed: 34990248
Uxa, S. et al. Ki-67 gene expression. Cell Death Differentiat. 28, 3357–3370 (2021).
doi: 10.1038/s41418-021-00823-x
Pan, C. et al. Shrinkage-mediated imaging of entire organs and organisms using uDISCO. Nat. Methods 13, 859–867 (2016).
doi: 10.1038/nmeth.3964
pubmed: 27548807
Jing, D. et al. Tissue clearing of both hard and soft tissue organs with the PEGASOS method. Cell Res. 28, 803–818 (2018).
doi: 10.1038/s41422-018-0049-z
pubmed: 29844583
pmcid: 6082844
Kosmidis, S., Negrean, A., Dranovsky, A., Losonczy, A. & Kandel, E. R. A fast, aqueous, reversible three-day tissue clearing method for adult and embryonic mouse brain and whole body. Cell Rep. Meth. 1, 100090 (2021).
doi: 10.1016/j.crmeth.2021.100090
Susaki, E. A. et al. Versatile whole-organ/body staining and imaging based on electrolyte-gel properties of biological tissues. Nat. Commun. 11, 1982 (2020).
doi: 10.1038/s41467-020-15906-5
pubmed: 32341345
pmcid: 7184626
Goc, J., Fridman, W. H., Sautès-Fridman, C. & Dieu-Nosjean, M. C. Characteristics of tertiary lymphoid structures in primary cancers. Oncoimmunology 2, e26836 (2013).
doi: 10.4161/onci.26836
pubmed: 24498556
pmcid: 3912008
Fletcher, A. L., Acton, S. E. & Knoblich, K. Lymph node fibroblastic reticular cells in health and disease. Nat. Rev. Immunol. 15, 350–361 (2015).
doi: 10.1038/nri3846
pubmed: 25998961
pmcid: 5152733
Takemura, S. et al. Lymphoid neogenesis in rheumatoid synovitis1. J. Immunol. 167, 1072–1080 (2001).
doi: 10.4049/jimmunol.167.2.1072
pubmed: 11441118
Srikakulapu, P. & McNamara, C. A. B cells and atherosclerosis. Am. J. Physiol. Heart. Circ. Physiol. 312, H1060–H1067 (2017).
doi: 10.1152/ajpheart.00859.2016
pubmed: 28314764
pmcid: 5451581
Magliozzi, R. et al. A gradient of neuronal loss and meningeal inflammation in multiple sclerosis. Ann. Neurol. 68, 477–493 (2010).
doi: 10.1002/ana.22230
pubmed: 20976767
Boergens, K. M. et al. webKnossos: efficient online 3D data annotation for connectomics. Nat. Methods 14, 691–694 (2017).
doi: 10.1038/nmeth.4331
pubmed: 28604722