Abstract
Cell membrane is made up of a complex structure of lipids and proteins that diffuse laterally giving rise to what we call membrane fluidity. During cellular development, such as differentiation cell membranes undergo dramatic fluidity changes induced by proteins such as ARC and Cofilin among others. In this study we used the generalized polarization (GP) property of fluorescent probe Laurdan using two-photon microscopy to determine membrane fluidity as a function of time and for various cell lines. A low GP value corresponds to a higher fluidity and a higher GP value is associated with a more rigid membrane. Four different cell lines were monitored such as hN2, NIH3T3, HEK293 and L6 cells. Membrane fluidity was measured at 12h, 72h and 92 h. Our results show significant changes in membrane fluidity among all cell types at different time points. GP values tend to increase significantly within 92 h in hN2 cells and 72 h in NIH3T3 cells and only at 92 h in HEK293 cells. L6 showed a marked decrease in membrane fluidity at 72 h and starts to increase at 92 h. As expected, NIH3T3 cells have more rigid membrane at earlier time points. On the other hand, neurons tend to have the highest membrane fluidity at early time points emphasizing its correlation with plasticity and the need for this malleability during differentiation. This study sheds light on the involvement of membrane fluidity during neuronal differentiation and development of other cell lines.
Highlights
Membrane fluidity is one of the major macroscopic biophysical properties characterizing cellular membranes
In L6 cells there was a significant decrease in generalized polarization (GP) values at 72 h compared to 12 h and a further increase at 92 h compared to 72 h (Fig 2D) (L6 12 h vs 72 h P = 6.93186E-12); (L6 12 h vs 92 h P = 1.69026E-08)
The variations in GP values among different cell lines studied above can be an indication of the difference in their lipid composition
Summary
Membrane fluidity is one of the major macroscopic biophysical properties characterizing cellular membranes. Several studies pointed at its implication during cellular events such as endocytosis, membrane fusion and importantly during development [1,2,3,4,5,6,7,8,9,10,11]. Differentiation of stem cells in vitro resembles to a certain extent embryo development. Lipids such as ceramide are involved in stem cell differentiation [12]. During this process prominent morphological changes occur.
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