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      Monitoring membrane viscosity in differentiating stem cells using BODIPY-based molecular rotors and FLIM

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          Abstract

          Membrane fluidity plays an important role in many cell functions such as cell adhesion, and migration. In stem cell lines membrane fluidity may play a role in differentiation. Here we report the use of viscosity-sensitive fluorophores based on a BODIPY core, termed “molecular rotors”, in combination with Fluorescence Lifetime Imaging Microscopy, for monitoring of plasma membrane viscosity changes in mesenchymal stem cells (MSCs) during osteogenic and chondrogenic differentiation. In order to correlate the viscosity values with membrane lipid composition, the detailed analysis of the corresponding membrane lipid composition of differentiated cells was performed by time-of-flight secondary ion mass spectrometry. Our results directly demonstrate for the first time that differentiation of MSCs results in distinct membrane viscosities, that reflect the change in lipidome of the cells following differentiation.

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          Restriction of receptor movement alters cellular response: physical force sensing by EphA2.

          Khalid Salaita, Pradeep Nair, Rebecca Petit … (2010)
          Activation of the EphA2 receptor tyrosine kinase by ephrin-A1 ligands presented on apposed cell surfaces plays important roles in development and exhibits poorly understood functional alterations in cancer. We reconstituted this intermembrane signaling geometry between live EphA2-expressing human breast cancer cells and supported membranes displaying laterally mobile ephrin-A1. Receptor-ligand binding, clustering, and subsequent lateral transport within this junction were observed. EphA2 transport can be blocked by physical barriers nanofabricated onto the underlying substrate. This physical reorganization of EphA2 alters the cellular response to ephrin-A1, as observed by changes in cytoskeleton morphology and recruitment of a disintegrin and metalloprotease 10. Quantitative analysis of receptor-ligand spatial organization across a library of 26 mammary epithelial cell lines reveals characteristic differences that strongly correlate with invasion potential. These observations reveal a mechanism for spatio-mechanical regulation of EphA2 signaling pathways.
          Article 0 comments Cited 91 times – based on 0 reviews     Review now
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            Sphingolipids--the enigmatic lipid class: biochemistry, physiology, and pathophysiology.

            Paul R. Riley, K.-O. Voss, J Shayman … (1996)
            The "sphingosin" backbone of sphingolipids was so named by J. L. W. Thudichum in 1884 for its enigmatic ("Sphinx-like") properties. Although still an elusive class of lipids, research on the involvement of sphingolipids in the signal transduction pathways that mediate cell growth, differentiation, multiple cell functions, and cell death has been rapidly expanding our understanding of these compounds. In addition to the newly discovered role of ceramide as an intracellular second messenger for tumor necrosis factor-alpha, IL-1beta, and other cytokines, sphingosine, sphingosine-1-phosphate, and other sphingolipid metabolites have recently been demonstrated to modulate cellular calcium homeostasis and cell proliferation. Perturbation of sphingolipid metabolism using synthetic and naturally occurring inhibitors of key enzymes of the biosynthetic pathways is aiding the characterization of these processes; for examples, inhibition of cerebroside synthase has indicated a role for ceramide in cellular stress responses including heat shock, and inhibition of ceramide synthase (by fumonisins) has revealed the role of disruption of sphingolipid metabolism in several animal diseases. Fumonisins are currently the focus of a FDA long-term tumor study. This review summarizes recent research on (i) the role of sphingolipids as important components of the diet, (ii) the role of sphingoid base metabolites and the ceramide cycle in expression of genes regulating cell growth, differentiation, and apoptosis, (iii) the use of cerebroside synthase inhibitors as tools for understanding the role of sphingolipids as mediators of cell cycle progression, renal disease, and stress responses, and (iv) the involvement of disrupted sphingolipid metabolism in animal disease and cellular deregulation associated with exposure to inhibitors of ceramide synthase and serine palmitoyltransferase, key enzymes in de novo sphingolipid biosynthesis. These findings illustrate how an understanding of the function of sphingolipids can help solve questions in toxicology and this is undoubtedly only the beginning of this story.
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              Lipid imaging with time-of-flight secondary ion mass spectrometry (ToF-SIMS).

              Nicholas Winograd, Melissa K. Passarelli (2011)
              Fundamental advances in secondary ion mass spectrometry (SIMS) now allow for the examination and characterization of lipids directly from biological materials. The successful application of SIMS-based imaging in the investigation of lipids directly from tissue and cells are demonstrated. Common complications and technical pitfalls are discussed. In this review, we examine the use of cluster ion sources and cryogenically compatible sample handling for improved ion yields and to expand the application potential of SIMS. Methodological improvements, including pre-treating the sample to improve ion yields and protocol development for 3-dimensional analyses (i.e. molecular depth profiling), are also included in this discussion. New high performance SIMS instruments showcasing the most advanced instrumental developments, including tandem MS capabilities and continuous ion beam compatibility, are described and the future direction for SIMS in lipid imaging is evaluated. Copyright © 2011 Elsevier B.V. All rights reserved.
              Article 0 comments Cited 73 times – based on 0 reviews     Review now
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                Author and article information

                Contributors
                Aleksandra V. Meleshina: almele@yandex.ru
                Marina K. Kuimova: m.kuimova@imperial.ac.uk
                Journal
                Journal ID (nlm-ta): Sci Rep
                Journal ID (iso-abbrev): Sci Rep
                Title: Scientific Reports
                Publisher: Nature Publishing Group UK (London )
                ISSN (Electronic): 2045-2322
                Publication date (Electronic): 20 August 2020
                Publication date PMC-release: 20 August 2020
                Publication date Collection: 2020
                Volume: 10
                Electronic Location Identifier: 14063
                Affiliations
                [1 ]Privolzhsky Research Medical University, 10/1 Minin and Pozharsky Sq., Nizhny Novgorod, Russian Federation 603950
                [2 ]GRID grid.7445.2, ISNI 0000 0001 2113 8111, Department of Chemistry, , Imperial College London, Molecular Sciences Research Hub, ; White City Campus, London, W12 0BZ UK
                [3 ]GRID grid.4886.2, ISNI 0000 0001 2192 9124, N.N. Semenov Federal Research Center for Chemical Physics, , Russian Academy of Sciences (FRCCP RAS), ; Kosygin st. 4, Moscow, Russian Federation 119991
                [4 ]GRID grid.14476.30, ISNI 0000 0001 2342 9668, Department of Chemistry, , Lomonosov Moscow State University, ; Leninskiye Gory 1-3, Moscow, Russian Federation 119991
                [5 ]Lobachevsky State University of Nizhny Novgorod, 23 Gagarin Avenue, Novgorod, Nizhny Novgorod, Russian Federation 603950
                Article
                Publisher ID: 70972
                DOI: 10.1038/s41598-020-70972-5
                PMC ID: 7441180
                PubMed ID: 32820221
                SO-VID: a5f79b4e-8560-4ace-8b19-2a6703b43901
                Copyright © © The Author(s) 2020
                License:

                Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

                History
                Date received : 3 April 2020
                Date accepted : 29 July 2020
                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/501100006769, Russian Science Foundation;
                Award ID: 17-75-20178
                Award ID: 17-75-20178
                Award ID: 17-75-20178
                Award ID: 17-75-20178
                Award ID: 17-75-20178
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/501100002261, Russian Foundation for Basic Research;
                Award ID: 18-33-00940
                Award Recipient :
                Categories
                Subject: Article
                Custom metadata
                issue-copyright-statement © The Author(s) 2020

                ScienceOpen disciplines: Uncategorized
                Keywords: stem cells,mesenchymal stem cells,cellular imaging
                Data availability:
                ScienceOpen disciplines: Uncategorized
                Keywords: stem cells, mesenchymal stem cells, cellular imaging

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