[Show abstract][Hide abstract] ABSTRACT: Our study emphasizes the effect of gamma irradiation on intestinal cell membrane fluidity and addresses the potential relationships existing between radiation-induced lipoperoxidation, membrane fluidity, and changes in membrane protein activities. Male Wistar rats were exposed to an 8-Gy total body irradiation (60Co source) and studied 1, 4, and 7 days after irradiation (D1, D4, and D7). Membrane enzyme activities and fluorescence anisotropy were determined on small intestinal crude membrane preparations. The supernatants of membrane preparations as well as plasma were used for malonedialdehyde (MDA) quantification. The effect of carbamylcholine on electrical parameters was estimated on distal ileum placed in Ussing chambers. We observed a decrease in fluorescence anisotropy for at least 7 days, an increase in membrane production of MDA at D4, a decrease in membrane enzyme activities at D4, but an amplification of carbamylcholine-induced increase in short-circuit current at D4 and D7. Furthermore, correlations were observed between the 1,6-diphenyl-1,3,5-hexatriene anisotropy coefficient and sucrase activity and between MDA levels and leucine aminopeptidase activity. Thus, total body irradiation induces changes in intestinal membrane fluidity and an increase in lipoperoxidation. These modifications may have an impact on the activity of membrane proteins involved in intestinal function.
Canadian Journal of Physiology and Pharmacology 08/2002; 80(7):686-93. DOI:10.1139/y02-091 · 1.77 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Modifications of intracellular transfer, resulting from a loss of membrane integrity may contribute toward setting the cell onto the pathway of apoptosis.
We have developed an original technique of measuring simultaneously, with flow cytometry, changes in membrane fluidity and cell death status. Our aim was to assess the extent to which radio-induced cell death and membrane alterations are linked. Investigations were performed on lymphocytes 24 h after whole human blood gamma-irradiation.
Our results confirmed the expected increase in the percentage of apoptotic cells as a function of dose, but revealed that the percentage of necrotic cells appeared stable after irradiation. At the same time, the fluorescence anisotropy of the living lymphocyte subpopulation decreased significantly and dose dependently as measured 24 h post-irradiation. With TMA-DPH, the anisotropy index of apoptotic lymphocytes was always lower than that of the viable lymphocyte subpopulation. On the other hand, 1,6-diphenyl-1,3,5-hexatriene (DPH) anisotropy was similar in apoptotic and viable cells after irradiation. These findings suggest that apoptotic lymphocytes are characterised by a membrane fluidization that mainly occurs on the cell membrane surface.
Our study made technical advances in using cytometric fluorescence anisotropy measurement as an early biological indicator of apoptosis after cellular exposure to ionising radiation.
[Show abstract][Hide abstract] ABSTRACT: This study focused on radio-induced membrane alterations in order to assess some related parameters as potential biological indicators of ionizing radiation effects in cases of accidental overexposure.
Radio-induced membrane alterations were assessed after gamma-irradiation of human blood. Biophysical techniques based on fluorescent probe incorporation into isolated living lymphocytes and erythrocytes membranes were applied.
Using the technique of fluorescence polarization, the lipophilic phase of the membrane was shown to be more fluid whereas the lipid-protein interface of the membrane was shown to be more rigid after gamma-irradiation. Fluorescent anisotropy modifications showed dose-time effect relationships after radiation exposure. Ionizing radiation induced a decrease in steady-state anisotropy values but did not affect the probe's lifetime as assessed by fluorescence lifetime distribution technique. These data suggest that the anisotropy variations are representative of the local properties of the fluorescent probe's micro-environment. However, the distribution width showed a decrease pointing towards radiation-induced changes of membrane domain organization, probably due not only to membrane water penetration related to lipoperoxidation, but also to compositional changes and redistribution of membrane components. In contrast, the lack of radiation effect observed using the lateral diffusion index technique may be related to the integrated overview of the radio-induced modifications of the membrane provided by this technique, which pointed out radio-induced damage to the membrane in micro-domains.
These findings suggest the utility of structural membrane modification measurements as an early bio-indicator of ionizing radiation exposure.
International Journal of Radiation Biology 09/1999; 75(8):1043-53. DOI:10.1080/095530099139818 · 1.69 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Effects of ionizing radiation on biological membranes include alterations in membrane proteins, peroxidation of unsaturated lipids accompanied by perturbations of the lipid bilayer polarity. We have measured radiation-induced membrane modifications using two fluorescent lipophilic membrane probes (TMA-DPH and DPH) by the technique of fluorescence polarization on two different cell lines (Chinese hamster ovary CHO-K1 and lymphoblastic RPMI 1788 cell lines). gamma-Irradiation was performed using a 60Co source with dose rates of 0.1 and 1 Gy/min for final doses of 4 and 8 Gy. Irradiation induced a decrease of fluorescence intensity and anisotropy of DPH and TMA-DPH in both cell lines, which was dose-dependent but varied inversely with the dose rate. Moreover, the fluorescence anisotropy measured in lymphoblastic cells using TMA-DPH was found to decrease as early as 1 h after irradiation, and remained significantly lower 24 h after irradiation. This study indicates that some alterations of membrane fluidity are observed after low irradiation doses and for some time thereafter. The changes in membrane fluidity might reflect oxidative damage, thus confirming a radiation-induced fluidization of biological membranes. The use of membrane fluidity changes as a potential biological indicator of radiation injury is discussed.