Laurdan monitors different lipids content in eukaryotic membrane during embryonic neural development.
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.
Department of Drug Science, Section of Biochemistry, University of Catania, Catania, Italy, gabriele.bonaventura@gmail.com.
Gabriele
G
Bonaventura
Maria Luisa
ML
Barcellona
Ottavia
O
Golfetto
Jamison L
JL
Nourse
Lisa A
LA
Flanagan
Enrico
E
Gratton
D015081:2-Naphthylamine / Q000031:analogs & derivatives* / Q000737:chemistry; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism*; D002784:Cholesterol / Q000378:metabolism; D047108:Embryonic Development; D005260:Female; D005456:Fluorescent Dyes / Q000737:chemistry*; D007848:Laurates / Q000737:chemistry*; D050356:Lipid Metabolism; D008560:Membrane Fluidity; D051379:Mice; D008856:Microscopy, Fluorescence / Q000379:methods*; D041681:NIH 3T3 Cells; D009474:Neurons / Q000166:cytology*; D011247:Pregnancy; D013997:Time Factors
D005456:Fluorescent Dyes; D007848:Laurates; D002784:Cholesterol; D015081:2-Naphthylamine; C065580:laurdan