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Laurdan Monitors Different Lipids Content in Eukaryotic Membrane During Embryonic Neural Development

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Abstract

We describe a method based on fluorescence-lifetime imaging microscopy (FLIM) to assess the fluidity of various membranes in neuronal cells at different stages of development [day 12 (E12) and day 16 (E16) of gestation]. For the FLIM measurements, we use the Laurdan probe which is commonly used to assess membrane water penetration in model and in biological membranes using spectral information. Using the FLIM approach, we build a fluidity scale based on calibration with model systems of different lipid compositions. In neuronal cells, we found a marked difference in fluidity between the internal membranes and the plasma membrane, being the plasma membrane the less fluid. However, we found no significant differences between the two cell groups, E12 and E16. Comparison with NIH3T3 cells shows that the plasma membranes of E12 and E16 cells are significantly more fluid than the plasma membrane of the cancer cells.

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Acknowledgments

Funding was provided by National Institutes of Health P50 GM076516, 5P41RR003155-27-8 P41 GM103540-27 (EG and OG), and UL1 TR000153 from the National Center for Research Resources (NCRR), a component of the National Institutes of Health (NIH) and the NIH Roadmap for Medical Research. GB and MB acknowledge funds from the Italian Ministry of University and Research MIUR 2012/13 PhD Neurobiology, Department of Chemical Sciences, University of Catania, Italy.

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Authors and Affiliations

  1. Department of Drug Science, Section of Biochemistry, University of Catania, Catania, Italy

    Gabriele Bonaventura & Maria Luisa Barcellona

  2. Laboratory for Fluorescence Dynamics, Department of Biomedical Engineering, University of California, Irvine, California

    Ottavia Golfetto & Enrico Gratton

  3. Department of Neurology, School of Medicine, University of California, Irvine, California

    Jamison L. Nourse & Lisa A. Flanagan

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  1. Gabriele Bonaventura
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  2. Maria Luisa Barcellona
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  3. Ottavia Golfetto
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  4. Jamison L. Nourse
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  5. Lisa A. Flanagan
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  6. Enrico Gratton
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Correspondence to Gabriele Bonaventura.

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Bonaventura, G., Barcellona, M.L., Golfetto, O. et al. Laurdan Monitors Different Lipids Content in Eukaryotic Membrane During Embryonic Neural Development. Cell Biochem Biophys 70, 785–794 (2014). https://doi.org/10.1007/s12013-014-9982-8

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