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Polyene-lipids: A new tool to image lipids

Nature Methods volume 2, pages 39–45 (2005)Cite this article

Abstract

Microscopy of lipids in living cells is currently hampered by a lack of adequate fluorescent tags. The most frequently used tags, NBD and BODIPY, strongly influence the properties of lipids, yielding analogs with quite different characteristics. Here, we introduce polyene-lipids containing five conjugated double bonds as a new type of lipid tag. Polyene-lipids exhibit a unique structural similarity to natural lipids, which results in minimal effects on the lipid properties. Analyzing membrane phase partitioning, an important biophysical and biological property of lipids, we demonstrated the superiority of polyene-lipids to both NBD- and BODIPY-tagged lipids. Cells readily take up various polyene-lipid precursors and generate the expected end products with no apparent disturbance by the tag. Applying two-photon excitation microscopy, we imaged the distribution of polyene-lipids in living mammalian cells. For the first time, ether lipids, important for the function of the brain, were successfully visualized.

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Figure 1: A versatile procedure to introduce conjugated pentaene tags into lipids.
Figure 2: Analysis of phase partitioning of fluorescent SM analogs.
Figure 3: Comparison of lipid metabolites of polyene-labeled or radioactive fatty acids in COS7 cells.
Figure 4: Lipid metabolites of polyene–fatty acid in different cell types.
Figure 5: Imaging of fluorescent polyene-lipids in various types of living mammalian cells.
Figure 6: Localization of different lipid classes in living COS7 cells.

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Acknowledgements

We thank colleagues who kindly provided plasmids: P. Keller and J. White (ManII), M. Rolls (Sec61β) and R.E. Tsien (mRFP1). We thank M. Gruner and A. Rudolph for NMR analysis and K. Sandhoff for helpful suggestions. We acknowledge financial support by the Deutsche Forschungsgemeinschaft (SFB-TR 13, D2).

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Authors and Affiliations

  1. Max-Planck-Institute of Molecular Cell Biology and Genetics, Pfotenhauerstr. 108, Dresden, 01307, Germany

    Lars Kuerschner, Christer S Ejsing, Andrej Shevchenko, Kurt I Anderson & Christoph Thiele

  2. AstraZeneca R&D, Moelndal, 43183, Sweden

    Kim Ekroos

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  1. Lars Kuerschner
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  2. Christer S Ejsing
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Correspondence to Christoph Thiele.

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Supplementary information

Supplementary Fig. 1

Mass spectrometric analysis of cellular glycerophospholipids containing the t/c16:5-fatty acid. (PDF 2477 kb)

Supplementary Fig. 2

Molecular modeling structures of stearic acid, t18:5-fatty acid, c18:5-fatty acid, and oleic acid. (PDF 531 kb)

Supplementary Table 1

Pentaene lipids of COS7 cells labeled with t16:5- or c16:5-fatty acids. (PDF 22 kb)

Supplementary Methods (PDF 56 kb)

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Kuerschner, L., Ejsing, C., Ekroos, K. et al. Polyene-lipids: A new tool to image lipids. Nat Methods 2, 39–45 (2005). https://doi.org/10.1038/nmeth728

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