We describe a method based on fluorescence lifetime imaging microscopy (FLIM)

We describe a method based on fluorescence lifetime imaging microscopy (FLIM) to assess the fluidity of various membranes in neuronal cells at different stage of development (day time 12 (E12) and day time 16 (E16) of gestation). the plasma membranes of E12 and E16 cells are significantly more fluid than the plasma membrane of the malignancy cells. Intro Membrane fluidity changes are implicated in a range of biological processes including signaling membrane fusion endocytosis and many others. Although the part of membrane fluidity during development has been discussed (1-10) a systematic study of changes in membrane fluidity during embryo development has not been SDZ 205-557 HCl carried out. Lipids and lipids domains play a fundamental role in the structural corporation of the cytoplasmatic membrane of eukaryotic cells. Lipids in biological membranes are fundamental for the boundary functions of cells including stimuli to growth and to immunological and stress response i.e. info delivered from the environment to the cell interior. Membranes of internal organelles allow the compartmentalization of cell functions. The complexity of the membrane lipid composition has suggested the coexistence of domains characterized by different dynamical properties in the membrane aircraft as sites for any putative preferential partitioning of proteins and solutes for modulating membrane activity and for diffusion along the aircraft and through the bilayer (11-19). In the last two decades studies on mind lipids have unequivocally demonstrated that many lipids have essential cell signaling functions (20-26). They SDZ 205-557 HCl are called ��bioactive lipids��. Lipid microdomains display high affinity to specific cell signaling proteins such as growth factors or cytokine receptors which lead to clustering and activation of these receptors (27-34). These domains are areas in the cell membrane (or intracellular membranes) that arise from your self-assembly of lipids in an ordered (Lo) structure in the liquid phase of the membrane. The lipid raft hypothesis has had a broad impact on the field of cellular biology especially in neural cells as practical membrane domains for cell-cell relationships and signal transduction (20-26). The minimum Rabbit Polyclonal to TUBGCP3. lipid SDZ 205-557 HCl composition of a raft is definitely cholesterol and sphingomyelin which is associated with glycerolsphingolipids (GSLs) or having a glycerolphospholipid such as phosphatidylcholine. Glycerophospholipids cholesterol and the sphingolipid backbone ceramide are synthesized in the ER. Considering the bidirectional vesicular connection between the ER and the plasma membrane the most impressive feature of the lipid corporation in mammalian cells is the enrichment of sphingolipids and cholesterol in the late Golgi plasma membrane and endosomes. Cholesterol spontaneously techniques between and across membranes like a monomer. Its location is determined by its high affinity for sphingolipids and saturated glycerophospholipids. The composition and concentration of gangliosides or sialic acid-containing GSLs switch dramatically during central nervous system (CNS) development. This switch in the composition of gangliosides is definitely correlated with defined developmental events and is evolutionarily conserved among many mammalian varieties (35-41). In general during CNS development the composition of GSLs begins with a relatively simple pattern with GM3 and GD3 predominating in early neuroectorderm. This pattern is definitely soon followed by the transient appearance of c-series gangliosides during the period of neural tube SDZ 205-557 HCl formation followed by a more complex pattern with four gangliosides of the a- and b-series GM1 GD1a GD1b and GT1b. These second option complex gangliosides constitute the major gangliosides in mature mind (35-41). Subcellular localization studies exposed that GM1 and GD1a are localized primarily within the plasma membrane and partly in the cytoplasm both as punctate clusters. This punctate distribution suggests localization of GM1 and/or GD1a in specialized structures such as membrane microdomains. It has been suggested that in mammalian neural cells membrane lipid rafts could serve as key assembly and sorting platforms for cell-cell relationships or transmission transduction complexes and modulate multiple cellular processes such as axonal growth and.