Very long chain fatty acids: Occurrence and biosynthesis in membrane fractions from etiolated maize coleoptiles

The endoplasmic reticulum from maize coleoptiles elongates stearoyl-CoA more effectively than the plasmalemma-enriched fraction. The alkane and very lo     

Characterization of the membrane-destabilizing properties of different pH-sensitive methacrylic acid copolymers

The intracellular delivery of active biomacromolecules from endosomes into the cytoplasm generally requires a membrane-disrupting agent. Since endosomes have a slightly acidic pH, anionic carboxylated polymers could be potentially useful for this purpose since they can destabilize membrane bilayers by pH-triggered conformational change. In this study, five different pH-sensitive methacrylic acid (MAA) copolymers were characterized with respect to their physicochemical and membrane lytic properties as a function of pH. pH-dependent conformational changes were studied in aqueous solution by turbidimetry and spectrofluorimetry. The hydrophobic domains that formed upon a decrease in pH were found to be dependent on copolymer's composition. Hemolysis and cytotoxicity assays demonstrated that the presence of the hydrophobic ethyl acrylate monomer and/or sufficient protonation of the carboxylic acid groups were important parameters for efficient membrane destabilization. Excessive copolymer hydrophobicity was not associated with membrane destabilization, but resulted in high macrophage cytotoxicity. Overall, this study gave more insights into the structure-activity relationship of MAA copolymers with membrane bilayers. Gaining knowledge of modulation of the physicochemical properties of copolymers and the optimization of copolymer-lipid interactions may lead to the elaboration of much more efficient drug delivery systems.     

Transport of Sterols to the Plasma Membrane of Leek Seedlings

To investigate the intracellular transport of sterols in etiolated leek (Allium porrum L.) seedlings, in vivo pulse-chase experiments with [1-¹⁴C]acetate were performed. Then, endoplasmic reticulum-, Golgi-, and plasma membrane (PM)-enriched fractions were prepared and analyzed for the radioactivity incorporated into free sterols. In leek seedlings sterols are present as a mixture in which (24R)-24-ethylcholest-5-en-3β-ol is by far the major compound (around 60%). The other sterols are represented by cholest-5-en-3β-ol, 24-methyl-cholest-5-en-3β-ol, (24S)-24-ethylcholesta-5,22E-dien-3β-ol, and stigmasta-5,24(24¹)Z-dien-3β-ol. These compounds are shown to reside mainly in the PM. Our results clearly indicate that free sterols are actively transported from the endoplasmic reticulum to the PM during the first 60 min of chase, with kinetics very similar to that of phosphatidylserine. Such a transport was found to be decreased at low temperature (12°C) and following treatment with monensin and brefeldin A. These data are consistent with a membrane-mediated process for the intracellular transport of sterols to the PM, which likely involves the Golgi apparatus.     

Application of proteomics to the assessment of the response to ionising radiation in Arabidopsis thaliana

Ionising radiation (IR) affects cellular and tissue function. However, the biological effects and interactions induced by IR are unclear. The aim of this study was to decipher the proteomic patterns that influence these pathways. The proteomes of Arabidopsis thaliana roots and rosettes were analysed in response to sub-lethal IR doses (0, 10, and 40 Gy). For each dose, the dynamic response was observed at different time points (2, 24 and 72 h). To quantitatively measure the effect of IR on the proteome, total proteins were extracted and subjected to 2-DE, and the changes in the 2-DE protein profiles were analysed. Statistical analysis revealed a total of 172 proteins (145 in leaves and 27 in roots) that were differentially expressed. These proteins were subsequently analysed by MALDI-TOF/TOF MS and comparative database analysis, and 144 (118 in leaves and 26 in roots) proteins were identified. The changes in the protein profile were quantitatively more significant for the 40 Gy dose than for the 10 Gy dose. In addition, specific functional groups of proteins were identified based on the consistency of the dose- and time-responses. The molecular mechanisms involved in the response to IR and a comparison of the observed responses are discussed.     

Lipid rafts in higher plant cells: purification and characterization of Triton X-100-insoluble microdomains from tobacco plasma membrane

A large body of evidence from the past decade supports the existence of functional microdomains in membranes of animal and yeast cells, which play important roles in protein sorting, signal transduction, or infection by pathogens. They are based on the dynamic clustering of sphingolipids and cholesterol or ergosterol and are characterized by their insolubility, at low temperature, in nonionic detergents. Here we show that similar microdomains also exist in plant plasma membrane isolated from both tobacco leaves and BY2 cells. Tobacco lipid rafts were found to be greatly enriched in a sphingolipid, identified as glycosylceramide, as well as in a mixture of stigmasterol, sitosterol, 24-methylcholesterol, and cholesterol. Phospho- and glycoglycerolipids of the plasma membrane were largely excluded from lipid rafts. Membrane proteins were separated by one- and two-dimensional gel electrophoresis and identified by tandem mass spectrometry or use of specific antibody. The data clearly indicate that tobacco microdomains are able to recruit a specific set of the plasma membrane proteins and exclude others. We demonstrate the recruitment of the NADPH oxidase after elicitation by cryptogein and the presence of the small G protein NtRac5, a negative regulator of NADPH oxidase, in lipid rafts.     

Laser ablation of the sonic hedgehog-a-expressing cells during fin regeneration affects ray branching morphogenesis

The zebrafish fin is an excellent system to study the mechanisms of dermal bone patterning. Fin rays are segmented structures that form successive bifurcations both during ontogenesis and regeneration. Previous studies showed that sonic hedgehog (shha) may regulate regenerative bone patterning based on its expression pattern and functional analysis. The present study investigates the role of the shha-expressing cells in the patterning of fin ray branches. The shha expression domain in the basal epidermis of each fin ray splits into two prior to ray bifurcation. In addition, the osteoblast proliferation profile follows the dynamic expression pattern of shha. A zebrafish transgenic line, 2.4shh:gfpABC#15, in which GFP expression recapitulates the endogenous expression of shha, was used to specifically ablate shha-expressing cells with a laser beam. Such ablations lead to a delay in the sequence of events leading to ray bifurcation without affecting the overall growth of the fin ray. These results suggest that shha-expressing cells direct localized osteoblast proliferation and thus regulate branching morphogenesis. This study reveals the fin ray as a new accessible system to investigate epithelial-mesenchymal interactions leading to organ branching.     

Enzymatic variability in the cattail (Typha angustifolia andT. latifolia,Typhaceae)

The enzymatic spectra ofTypha angustifolia, T. latifolia and their hybrids were examined by disc electrophoresis for three enzymatic systems: esterases, alcohol deshydrogenase, and glutamate deshydrogenase. The intermediate position of the hybrids is confirmed. The study of alcohol deshydrogenase reveales the existence of an intraspecific variability inT. angustifolia between individuals from the Massif Armoricain in France and Dellys in Algeria.     

Simple standard procedure for the routine determination of organic matter in marine sediment

For the routine determination of organic matter inmarine sediment, a rough estimate is often obtained bymeasuring the loss of weight on ignition(Difference-On-Ignition method: D.O.I.). This D.O.I. is to beused in ecological studies where organic material insediment is used as an environmental variable amongothers. A review of papers using this method showed agreat variability within conditions used. Theseconditions are generally used without accuratejustification. We propose a simple standard procedurebased on grain-size fraction of sediment to determineoptimal temperature and time of ignition, and theinfluence of the sample weight. The method proposed, using the loss of weight on ignition, will give thesame accuracy in measurements of organic matter inmarine sediments.