The fluorescent probe 1,6-diphenyl-1,3,5-hexatriene (DPH) and DPH derivatives have been used to characterize structural and physicochemical properties of specific membrane domains. Steady-state and fluorescence decay measurements of three probes, DPH (1,6-diphenyl-1,3,5-hexatriene), TMA-DPH [1-(4-trimethyl-ammonium-phenyl)-6-phenyl-1,3,5-hexatriene], and a phosphatidylcholine derivative of DPH, DPH-pPC [2-(3-(diphenylhexatriene)propanoyl)-3-pamitoyl-L-α-phosphatidyl choline], have been performed in erythrocyte membranes and in lymphocyte plasma membranes. The steady-state fluorescence polarization of the three probes showed a similar trend in both membranes.
View Article and Find Full Text PDFThe interaction between lipoproteins and the platelet membrane has been proved to cause a modification in cellular functions. We studied 12 men with insulin-dependent diabetes mellitus (IDDM), 14 men with noninsulin-dependent diabetes mellitus (NIDDM), and 26 age-matched healthy men on the same diet. We determined fluidity by measuring the fluorescence polarization (P) of the probe 1,6-diphenyl-1,3,5-hexatriene (DPH) both in platelet membranes and in lipoproteins isolated by ultracentrifugation in NaBr density gradient.
View Article and Find Full Text PDFHepatic ethanol (EtOH) metabolism has been assumed to involve hepatocytes differently, according to their location in the hepatic acinus. This study's aim was to gain information on plasma membrane (PM) order parameter in periportal (PP) and perivenular (PV) hepatocyte-enriched fractions isolated by a digitonin-collagenase perfusion technique from rats pair-fed for 6-8 wk liquid diets containing either EtOH or isocaloric carbohydrates. Fluorescence polarization (P) studies have been performed to measure PM order parameter of PP and PV hepatocytes cultured for 2-6 h on glass cover slips and labeled with 1-[4-(trimethylamino)phenyl]-6-phenylhexa-1,3,5-triene (TMA-DPH), a specific probe for PM of living cells.
View Article and Find Full Text PDFUsing static and dynamic fluorescence we studied the fluorescence properties of a phosphatidylcholine analog of 1,6-diphenyl-1,3,5-hexatriene (DPH-PC) incorporated in lymphocyte plasma membranes with respect to DPH and its cationic derivative 1-(4-trimethylammoniumphenyl)-6-phenyl-1,3,5-hexatriene (TMA-DPH), in order to study if phospholipid derivatives of DPH may be used to investigate structural and physicochemical properties of specific membrane lipid domains. DPH-PC and TMA-DPH showed similar fluorescence polarization values that were significantly higher with respect to DPH, suggesting a localization of the fluorescent portion of DPH-PC in a more ordered region of the membrane which was probably due to the elecrostatic interactions between phospholipid head-groups. The localization of the fluorescent moiety of DPH-PC near the membrane surface was also supported by the study of the fluorescence decay of the three probes using frequency-domain fluorometry.
View Article and Find Full Text PDFIn this study we have demonstrated a transfer of a phosphatidylcholine derivative of 1,6-diphenyl-1,3,5-hexatriene (DPH-PC) from self-quenched lipid vesicles to intact lymphocytes. Membrane labeling was followed measuring the time dependent reexpression of fluorescence. The results of fluorescence quenching by 2,4,6-trinitrobenzene sulfonate and the decrease of fluorescence polarization values during incubation at 37 degrees C, suggest that the probe could remain localized at level of the plasma membrane until 20-30 minutes.
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