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At least 109 records · Page 6Linked to original sources

High-resolution proton NMR measures mobile lipids associated with Triton-resistant membrane domains in haematopoietic K562 cells lacking or expressing caveolin-1.

High-resolution proton NMR spectra of intact tumour cells generally exhibit intense signals due to isotropically mobile lipids (MLs) of still uncertain nature and origin. NMR studies performed on intact wild-type and caveolin-1-infected haematopoietic K562 cells showed that, under our experimental conditions, part of the ML signals are due to lipid complexes resistant to extraction in Triton X-100 at 4 degrees C. This evidence suggests that a portion of NMR-visible lipid structures are compatible with Triton-resistant membrane rafts and therefore biophysically distinct from NMR-visible Triton-soluble lipid bodies. Similarly to lipid rafts and caveolae, the organization of the Triton-insoluble ML domains could be compromised by treatment with beta-octylglucoside or methyl-beta-cyclodextrin. Exposure to exogenous sphingomyelinase caused an increase in ML NMR visibility, indicating the possible involvement of ceramides in ML formation. The mobility of these lipids was found to be temperature sensitive, suggesting a transition in cells going from 4 degrees C to 25-37 degrees C. These new results are here discussed in the light of possible contributions of plasma membrane microdomains to NMR-visible ML signals.

Caveolin 1↗

Radiation effects in cultured tumour cells examined by 1H MRS: mobile lipids modulation and proliferative arrest.

Much attention has been devoted in the past to monitor changes of mobile lipid (ML) (1)H MRS signals in spectra of tumour cells. The purpose of this work is to exploit ML signals to provide information on cell metabolism after irradiation, comparing tumour cells characterised by different radiosensitivity and relating MRS findings to changes in cell proliferation and delays in cell cycle phases. Irradiated HeLa cells present less intense ML signals with respect to controls. The opposite is true for MCF-7 cells. A G(2) arrest is observed for both cell lines after irradiation. In HeLa cells, G(1) decreases and S phase is maintained; a sub G(1) peak is also visible. In MCF-7 cells, G(1) is decreased and S phase is strongly reduced, while no sub G(1) is present. The observed changes in ML are tentatively associated to cell cycle regulation of phospholipid synthesis. Mathematical modelling of ML variations is in progress.

Cell Cycle↗

Alterations in lateral lipid mobility in the plasma membrane of urodelean ectodermal cells during gastrulation.

The mobility characteristics of lipids were studied in the plasmalemma of dissociated presumptive ectodermal cells from embryos of Pleurodeles Waltl at different stages of development, from early blastula to early neurula, using a Fluorescence Recovery After Photobleaching technique (FRAP), after incorporation of the lipophilic fluorescent probe 5N-(hexadecanoyl)-aminofluoresceine (HEDAF) into the cell plasma membrane. At all stages of development, fluorescence recovery was found to extrapolate to 100%, which suggested that the lipid phase in these plasma membranes can be regarded as dynamically homogeneous (no immobilized fraction). It appears as a continuum over a wide cell surface area, in which lipids are free to move laterally. The lateral diffusion coefficient of the probe, obtained from statistical analysis of the fluorescence recovery data, was found to decrease significantly from blastula to gastrula, slightly increasing at the neurula stage. These changes in the dynamic properties of the lipid probe HEDAF during gastrulation suggest that the lipid phase of the plasma membrane of these ectodermal cells undergo structural changes. The results lend support to the idea that the plasma membrane of these cells is actively involved in the morphogenetic movements which characterize the development of the embryo.

Animals↗

Effects of dogfish urotensin II on lipid mobilization in the fasted dogfish, Scyliorhinus canicula.

Previous studies, both in vitro and in vivo, have shown that urotensin II stimulates hepatic release of nonesterified fatty acids in teleost fish, but the effects of the peptide on lipid mobilization in elasmobranchs are unknown. Infusions of synthetic dogfish urotensin II (40 pmol/min/kg for 30 min followed by 400 pmol/min/kg for 30 min) into the first afferent gill artery of the fasted, unrestrained dogfish, Scyliorhinus canicula (n = 6), produced no significant change in the circulating concentrations of acetoacetate, 3-hydroxybutyrate, and triacylglycerols. Circulating nonesterified fatty acids were undetectable (< 0.1 mmol/liter) both before and after the urotensin II infusions. There was no significant change in the circulating concentrations of glucose and lactate during either the low- or the high-dose infusions. The data support the hypothesis that lipid is converted primarily to ketone bodies in the livers of elasmobranch fish and do not indicate a role for urotensin II in regulating either lipid or glycogen mobilization.

3-Hydroxybutyric Acid↗

Temperature-induced protein conformational changes in barley root plasma membrane-enriched microsomes: I. Effect of temperature on membrane protein and lipid mobility.

A protein spin label and lipid spin probes were used to study the temperature-dependent motion of protein and lipid, respectively, in barley (Hordeum vulgare L. cv Conquest) root plasma membrane-enriched microsomes. Using membranes from seedlings grown at 20 degrees C, the temperature-dependence of the relative motion of membrane surface spin probes and a spin label covalently attached to membrane proteins suggested abrupt changes in the lipid and protein mobilities at about 12 degrees C. Spin probe spin-spin exchange broadening and fluorescent probe eximer formation indicated apparent temperature-induced alterations in probe lateral diffusion within the membrane at about 12 to 14 degrees C. The results suggest the presence of temperature-induced quasicrystalline lipid clusters which may influence the activity of membrane-bound enzymes.

Journal Article↗

Two-step formation of 1H NMR visible mobile lipids during apoptosis of paclitaxel-treated K562 cells.

Despite increasing evidence on the formation of 1H NMR-detectable mobile lipid (ML) domains in cells induced to programmed cell death by continuous exposure to anticancer drugs, the time course of ML generation during the apoptotic cascade has not yet been fully elucidated. The present study shows that ML formation occurs at two different stages of apoptosis induced in human erythroleukemia K562 cells by a brief (3 hr) exposure to paclitaxel (Taxol), an antitumour drug with a stabilising effect on microtubules, or to paclitaxel plus tyrphostin AG957, a selective inhibitor of the p210(BCR-ABL) tyrosine kinase activity. A first wave of ML generation was in fact detected in paclitaxel-treated cells at the onset of the effector phase (8-24hr after exposure to the drug), plateaued at 24-48 hr and was eventually followed by further ML accumulation during the degradative phase (48-72 hr). Addition of AG957 to paclitaxel shifted to the 3-8 hr interval in both the early ML production and the onset of apoptotic events, such as chromatin condensation, phosphatidylserine externalization, cytochrome c release and caspase-3 activation. A significant loss of mitochondrial membrane potential was almost concomitant with the second wave of ML accumulation, associated in both cell systems with the phase of terminal cell degeneration, likely connected to non-regulated degradation of cell lipid components.

Annexin A5↗

Probing lipid mobility of raft-exhibiting model membranes by fluorescence correlation spectroscopy.

Confocal fluorescence microscopy and fluorescence correlation spectroscopy (FCS) have been employed to investigate the lipid spatial and dynamic organization in giant unilamellar vesicles (GUVs) prepared from ternary mixtures of dioleoyl-phosphatidylcholine/sphingomyelin/cholesterol. For a certain range of cholesterol concentration, formation of domains with raft-like properties was observed. Strikingly, the lipophilic probe 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI-C18) was excluded from sphingomyelin-enriched regions, where the raft marker ganglioside GM1 was localized. Cholesterol was shown to promote lipid segregation in dioleoyl-phosphatidylcholine-enriched, liquid-disordered, and sphingomyelin-enriched, liquid-ordered phases. Most importantly, the lipid mobility in sphingomyelin-enriched regions significantly increased by increasing the cholesterol concentration. These results pinpoint the key role, played by cholesterol in tuning lipid dynamics in membranes. At cholesterol concentrations >50 mol%, domains vanished and the lipid diffusion slowed down upon further addition of cholesterol. By taking the molecular diffusion coefficients as a fingerprint of membrane phase compositions, FCS is proven to evaluate domain lipid compositions. Moreover, FCS data from ternary and binary mixtures have been used to build a ternary phase diagram, which shows areas of phase coexistence, transition points, and, importantly, how lipid dynamics varies between and within phase regions.

Carbocyanines↗

Effects of high-dose medroxyprogesterone acetate and various other steroid hormones on plasma membrane lipid mobility in CAMA-1 mammary cancer cells.

The influence of medroxyprogesterone acetate (MPA) and various other steroid hormones on the lateral diffusion of the fluorescent lipid probe 1-acyl-2-(N-4-nitrobenzo-2-oxa-1.3-diazolyl)-aminocarpropylphos phatidylcholine (NBD-PC) in the plasma membrane of intact mammary cancer cells (CAMA-1 cell line) has been studied by fluorescence recovery after photobleaching technique. Incubation of cells with MPA in serum free medium at ambient temperature for 1 hr led to a significant decrease in the lateral diffusion coefficient of NBD-PC. MPA induced this change over a limited concentration range with 10(-7)-10(-5) mol/l near-maximal or maximal effects, and 10(-8) mol/l exhibiting no effect. Exposure of the CAMA-1 cells to 10(-7) mol/l MPA in undiluted serum induced a significant effect following 1.5 hr of treatment with no increase in effectiveness up to 4 hr of incubation. As compared to MPA, the other steroids tested were less effective or ineffective. The influence on lateral lipid mobility diminished as follows: MPA greater than progesterone greater than 5a-DHT approximately 17 beta-estradiol greater than dexamethasone, and roughly seems to parallel their lipid solubility as estimated by partition coefficients in an n-octanol-water system. Any involvement of classical steroid hormone receptors in the mechanism of membrane action could be excluded. Nongenomic steroid effects on the plasma membrane are assumed. As the structure and function of biomembranes are modulated by lipid-bilayer fluidity and membranes crucially participate in nearly all aspects of cell biology, it is concluded that direct interactions of MPA with membranes potentially play a role in the antitumor activity of the compound.

Algorithms↗

Lipid mobilization by obese and lean pigs infused with the beta-adrenergic agonist isoproterenol.

Generally obese and lean pigs in both the fed and fasted states were anesthetized and then acutely infused with increasing concentrations of the beta-adrenergic agonist isoproterenol. Plasma free fatty acid (FFA), blood glycerol, glucose and lactate, and heart rate were monitored during the infusion period. Data were reduced by estimating the parameters of the generalized logistic function (minimum, maximum, ED50 and slope) and subsequently analyzed to compare the lean and obese genotypes within nutritional state. Lactate data could not be fitted to this function because the upper asymptote was not approached during the experiment. The minimum plasma concentration of FFA tended (P less than .1) to be less in obese than in lean pigs. The maximum, ED50 and slope for the responses of FFA were similar for obese and lean pigs in fed pigs and in fasted pigs. In fed pigs, the minimum glycerol concentration was greater in obese than in lean pigs, and the ED50 for heart rate tended to be lower in lean than in obese pigs. All other estimated parameters for the variables were similar in fed obese and lean pigs. In fasted pigs, the maximum glucose concentration was greater in obese than in lean pigs. All other parameters for the variables were similar in fasted obese and lean pigs. The results suggest that there was no major defect in lipid mobilization in these obese pigs (only a lower minimum FFA concentration was detected) and that an increased maximum blood glucose concentration in the fasting state might contribute to the obesity.

Animals↗

[Lipid-mobilizing effect of reduced glutathione and its intensifying action on lecithin-cholesterol acyltransferase activity in blood serum of rats].

Effect of reduced glutathione (50 mg/100 g) on lipid distribution between organs (liver and kidney) and lecithin-cholesterol acyltransferase (LCAT) activity in blood serum of rats was investigated. The accumulation of common lipids as a result of speeding up the absorbtion of blood serum unsaturated fatty acids and relative decrease of lipids unsaturation in the liver and lipid content dynamics in kidneys owing to the intensification of two processes in this organ: the transport of polyene fatty acids in composition of blood serum lipoprotein lipids to kidney cells and peroxidation of membrane phospholipids were found out. The activating effect of GSH (in vivo and in vitro) on LCAT activity of rat blood serum was shown. It was summarised that GSH-intensification of blood serum etherification ability may be a basic component of reduced glutathione lipid mobilization effect.

Animals↗

Gender differences in adrenergic regulation of lipid mobilization during exercise.

Gender differences in adrenergic regulation of glycerol levels in subcutaneous, abdominal adipose tissue were investigated during submaximal exercise in non-obese, healthy men and women, using microdialysis. During exercise, glycerol levels in venous plasma and venous serum concentrations of free fatty acids increased more in women and reached about two-fold higher values than in men (p < 0.005 or less). Plasma noradrenaline and insulin did not differ between the sexes, whereas plasma adrenaline was two-fold higher in men than in women during exercise (p < 0.01). The glycerol levels in adipose tissue increased during exercise and decreased in the post-exercise period in either sex. When the non-selective beta adrenoceptor blocking agent propranolol was added to the microdialysis perfusate before exercise was initiated, the subsequent increase in dialysate glycerol was significantly diminished in both sexes (p < 0.05). A similar addition of the alpha adrenoceptor blocking agent phentolamine, however, caused a significant further rise in tissue glycerol in men (p < 0.05), whereas the exercise induced increase in glycerol levels remained unaffected by phentolamine in women. Adipose tissue blood flow did not change during exercise in either men or women. In either sex, dialysate lactate levels increased during exercise. This increase was not altered if alpha- or beta-blocking agents were added to the perfusate. In summary, during short term submaximal work, women have a higher increase in circulating lipid than men. This appears, at least in part, to be due to a sex difference in the adrenergic regulation of lipid mobilization during exercise. In men exercise activates beta- as well as alpha-adrenergic receptors in adipose tissue, whereas only beta receptors are activated in adipose tissue of women. Finally, methodological investigations indicate that microdialysis is a valid method for short-term exercise experiments.

Adipose Tissue↗

Increased rate of capping of concanavalin A receptors during early Xenopus development is related to changes in protein and lipid mobility.

The mobility characteristics of plasma membrane constituents were studied in dissociated cells from embryos of Xenopus laevis at various stages of development from early blastula until neurulation. An increased rate of fluorescein isothiocyanate-concanavalin A induced patching and capping of Con A-binding proteins during this period of development was correlated with a threefold increase in the lateral mobility of the receptor molecules, as determined by the fluorescent photobleaching recovery (FPR) method, the major change occurring at the onset of gastrulation. Using the same method, it was demonstrated that the lateral mobility of plasma membrane lipids increases twofold during this period of development. The major change being detectable, however, at the late blastula stage. This is in coincidence with the initiation of cell motility in dissociated Xenopus embryo cells. It is concluded that the lateral mobility of membrane proteins and lipids increases significantly during early Xenopus development, but are at least in part subject to different control mechanisms. The results suggest that the initiation of morphogenetic movements is related to changes in the dynamic properties of plasma membrane constituents.

Animals↗