Development of an Optimized Clearing Protocol to Examine Adipocyte Subpopulations in White Adipose Tissue
Abstract
1. Introduction
2. Materials and Methods
3. Results
3.1. BABB and BABB-D4 Clearing Increased Adipose Tissue Transparency and Reduced Tissue Shrinkage Compared to DBE-Based Clearing
3.2. Type-1 Adipocytes Are Spatially Clustered and Are Preferentially Concentrated in Mesenteric Fat Associated with the Ileum and Colon
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Ariel, P. A beginner’s guide to tissue clearing. Int. J. Biochem. Cell Biol. 2017, 84, 35–39. [Google Scholar] [CrossRef]
- Azaripour, A.; Lagerweij, T.; Scharfbillig, C.; Jadczak, A.E.; Willershausen, B.; Van Noorden, C.J. A survey of clearing techniques for 3D imaging of tissues with special reference to connective tissue. Prog. Histochem. Cytochem. 2016, 51, 9–23. [Google Scholar] [CrossRef] [PubMed]
- Ueda, H.R.; Ertürk, A.; Chung, K.; Gradinaru, V.; Chédotal, A.; Tomancak, P.; Keller, P.J. Tissue clearing and its applications in neuroscience. Nat. Rev. Neurosci. 2020, 21, 61–79. [Google Scholar] [CrossRef]
- Orlich, M.; Kiefer, F. A qualitative comparison of ten tissue clearing techniques. Histol Histopathol. 2018, 33, 181–199. [Google Scholar] [PubMed]
- Silvestri, L.; Costantini, I.; Sacconi, L.; Pavone, F.S. Clearing of fixed tissue: A review from a microscopist’s perspective. J. Biomed. Opt. 2016, 21, 081205. [Google Scholar] [CrossRef] [PubMed]
- Tchkonia, T.; Tchoukalova, Y.D.; Giorgadze, N.; Pirtskhalava, T.; Karagiannides, I.; Forse, R.A.; Koo, A.; Stevenson, M.; Chinnappan, D.; Cartwright, A.; et al. Abundance of two human preadipocyte subtypes with distinct capacities for replication, adipogenesis, and apoptosis varies among fat depots. Am. J. Physiol. Metab. 2005, 288, E267–E277. [Google Scholar] [CrossRef]
- Lee, K.Y.; Luong, Q.; Sharma, R.; Dreyfuss, J.M.; Ussar, S.; Kahn, C.R. Developmental and functional heterogeneity of white adipocytes within a single fat depot. EMBO J. 2018, 10, e99291. [Google Scholar] [CrossRef]
- Sanchez-Gurmaches, J.; Guertin, D.A. Adipocytes arise from multiple lineages that are heterogeneously and dynamically distributed. Nat. Commun. 2014, 5, 4099. [Google Scholar] [CrossRef]
- Sanchez-Gurmaches, J.; Hsiao, W.-Y.; Guertin, D.A. Highly Selective In Vivo Labeling of Subcutaneous White Adipocyte Precursors with Prx1-Cre. Stem Cell Rep. 2015, 4, 541–550. [Google Scholar] [CrossRef] [PubMed]
- Sebo, Z.L.; Jeffery, E.; Holtrup, B.; Rodeheffer, M.S. A mesodermal fate map for adipose tissue. Development 2018, 145, dev.166801. [Google Scholar] [CrossRef]
- Cao, Y.; Wang, H.; Wang, Q.; Han, X.; Zeng, W. Three-dimensional volume fluorescence-imaging of vascular plasticity in adipose tissues. Mol. Metab. 2018, 14, 71–81. [Google Scholar] [CrossRef]
- Jiang, H.; Ding, X.; Cao, Y.; Wang, H.; Zeng, W. Dense Intra-adipose Sympathetic Arborizations Are Essential for Cold-Induced Beiging of Mouse White Adipose Tissue. Cell Metab. 2017, 26, 686–692.e3. [Google Scholar] [CrossRef]
- Cao, Y.; Wang, H.; Zeng, W. Whole-tissue 3D imaging reveals intra-adipose sympathetic plasticity regulated by NGF-TrkA signal in cold-induced beiging. Protein Cell 2018, 9, 527–539. [Google Scholar] [CrossRef]
- Chi, J.; Wu, Z.; Choi, C.H.J.; Nguyen, L.; Tegegne, S.; Ackerman, S.E.; Crane, A.; Marchildon, F.; Tessier-Lavigne, M.; Cohen, P. Three-Dimensional Adipose Tissue Imaging Reveals Regional Variation in Beige Fat Biogenesis and PRDM16-Dependent Sympathetic Neurite Density. Cell Metab. 2018, 27, 226–236.e3. [Google Scholar] [CrossRef]
- Kolesová, H.; Čapek, M.; Radochová, B.; Janáček, J.; Sedmera, D. Comparison of different tissue clearing methods and 3D imaging techniques for visualization of GFP-expressing mouse embryos and embryonic hearts. Histochem. Cell Biol. 2016, 146, 141–152. [Google Scholar] [CrossRef] [PubMed]
- Muzumdar, M.; Tasic, B.; Miyamichi, K.; Li, L.; Luo, L. A global double-fluorescent Cre reporter mouse. Genes 2007, 45, 593–605. [Google Scholar] [CrossRef] [PubMed]
- Chi, J.; Crane, A.; Wu, Z.; Cohen, P. Adipo-Clear: A Tissue Clearing Method for Three-Dimensional Imaging of Adipose Tissue. J. Vis. Exp. 2018, e58271. [Google Scholar] [CrossRef]
- Pan, C.; Cai, R.; Quacquarelli, F.P.; Ghasemigharagoz, A.; Lourbopoulos, A.; Matryba, P.; Plesnila, N.; Dichgans, M.; Hellal, F.; Ertürk, A. Shrinkage-mediated imaging of entire organs and organisms using uDISCO. Nat. Methods 2016, 13, 859–867. [Google Scholar] [CrossRef] [PubMed]
- Botev, Z.I.; Grotowski, J.F.; Kroese, D.P. Kernel density estimation via diffusion. Ann. Stat. 2010, 38, 2916–2957. [Google Scholar] [CrossRef]
- Chang, W. 6.12 Making a Density Plot of Two-Dimensional Data. R Graphics Cookbook, 2nd ed. Available online: https://r-graphics.org (accessed on 23 April 2021).
- François, M.; Torres, H.; Huesing, C.; Zhang, R.; Saurage, C.; Lee, N.; Qualls-Creekmore, E.; Yu, S.; Morrison, C.D.; Burk, D.; et al. Sympathetic innervation of the interscapular brown adipose tissue in mouse. Ann. N. Y. Acad. Sci. 2019, 1454, 3–13. [Google Scholar] [CrossRef] [PubMed]
- Ertürk, A.; Becker, K.; Jährling, N.; Mauch, C.P.; Hojer, C.D.; Egen, J.G.; Hellal, F.; Bradke, F.; Sheng, M.; Dodt, H.-U. Three-dimensional imaging of solvent-cleared organs using 3DISCO. Nat. Protoc. 2012, 7, 1983–1995. [Google Scholar] [CrossRef] [PubMed]
- Renier, N.; Wu, Z.; Simon, D.J.; Yang, J.; Ariel, P.; Tessier-Lavigne, M. iDISCO: A Simple, Rapid Method to Immunolabel Large Tissue Samples for Volume Imaging. Cell 2014, 159, 896–910. [Google Scholar] [CrossRef]
- Dodt, H.-U.; Leischner, U.; Schierloh, A.; Jährling, N.; Mauch, C.P.; Deininger, K.; Deussing, J.; Eder, M.; Zieglgänsberger, W.; Becker, K. Ultramicroscopy: Three-dimensional visualization of neuronal networks in the whole mouse brain. Nat. Methods 2007, 4, 331–336. [Google Scholar] [CrossRef] [PubMed]
- Susaki, E.A.; Ueda, H.R. Whole-body and Whole-Organ Clearing and Imaging Techniques with Single-Cell Resolution: Toward Organism-Level Systems Biology in Mammals. Cell Chem. Biol. 2016, 23, 137–157. [Google Scholar] [CrossRef] [PubMed]
- Chau, Y.-Y.; Bandiera, R.; Serrels, A.; Martínez-Estrada, O.M.; Qing, W.; Lee, M.; Slight, J.; Thornburn, A.; Berry, R.; McHaffie, S.; et al. Visceral and subcutaneous fat have different origins and evidence supports a mesothelial source. Nat. Cell Biol. 2014, 16, 367–375. [Google Scholar] [CrossRef]
- Dobosz, M.; Ntziachristos, V.; Scheuer, W.; Strobel, S. Multispectral Fluorescence Ultramicroscopy: Three-Dimensional Visualization and Automatic Quantification of Tumor Morphology, Drug Penetration, and Antiangiogenic Treatment Response. Neoplasia 2014, 16, 1–13. [Google Scholar] [CrossRef] [PubMed]
- Chen, Y.; Shen, Q.; White, S.L.; Gokmen-Polar, Y.; Badve, S.; Goodman, L.J. Three-dimensional imaging and quantitative analysis in CLARITY processed breast cancer tissues. Sci. Rep. 2019, 9, 5624. [Google Scholar] [CrossRef]
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. |
© 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Luong, Q.V.; Israel, A.; Sharma, R.; Ussar, S.; Lee, K.Y. Development of an Optimized Clearing Protocol to Examine Adipocyte Subpopulations in White Adipose Tissue. Methods Protoc. 2021, 4, 39. https://doi.org/10.3390/mps4020039
Luong QV, Israel A, Sharma R, Ussar S, Lee KY. Development of an Optimized Clearing Protocol to Examine Adipocyte Subpopulations in White Adipose Tissue. Methods and Protocols. 2021; 4(2):39. https://doi.org/10.3390/mps4020039
Chicago/Turabian StyleLuong, Quyen Vi, Andreas Israel, Rita Sharma, Siegfried Ussar, and Kevin Y. Lee. 2021. "Development of an Optimized Clearing Protocol to Examine Adipocyte Subpopulations in White Adipose Tissue" Methods and Protocols 4, no. 2: 39. https://doi.org/10.3390/mps4020039
APA StyleLuong, Q. V., Israel, A., Sharma, R., Ussar, S., & Lee, K. Y. (2021). Development of an Optimized Clearing Protocol to Examine Adipocyte Subpopulations in White Adipose Tissue. Methods and Protocols, 4(2), 39. https://doi.org/10.3390/mps4020039