Increase In Cardiolipin Content Research Articles

Amino-nanopolystyrene exposures of oyster (Crassostrea gigas) embryos induced no apparent intergenerational effects

Early life stages (ELS) of numerous marine invertebrates mustcope with man-made contaminants, including plastic debris, during their pelagic phase. Among the diversity of plastic particles, nano-sized debris, known as nanoplastics, can induce effects with severe outcomes in ELS of various biological models, including the Pacific oyster Crassostrea gigas. Here, we investigated the effects of a sub-lethal dose (0.1 µg mL−1) of 50 nm polystyrene nanobeads (nano-PS) with amine functions on oyster embryos (24 h exposure) and we assessed consequences on larval and adult performances over two generations of oysters. Only a few effects were observed. Lipid analyses revealed that first-generation (G1) embryos exposed to nano-PS displayed a relative increase in cardiolipin content (+9.7%), suggesting a potential modification of mitochondrial functioning. G1-larvae issued from exposed embryos showed decreases in larval growth (−9%) and lipid storage (−20%). No effect was observed at the G1 adult stage in terms of growth, ecophysiological parameters (clearance and respiration rates, absorption efficiency), or reproductive outputs (gonadic development, gamete quality). Second generation (G2) larvae issued from control G1 displayed a significant growth reduction after G2 embryonic exposure to nano-PS (−24%) compared to control (as observed at the first generation), while no intergenerational effect was detected on G2 larvae issued from G1 exposed embryos. Overall, the present experimental study suggests a low incidence of a short embryonic exposure to nano-PS on oyster phenotypes along the entire life cycle until the next larval generation.

Bacillus subtilis cardiolipin protects its own membrane against surfactin-induced permeabilization

Surfactin, a cyclic lipoheptapeptide produced by Bacillus subtilis, is a surface-active antimicrobial that targets the barrier function of lipid membranes. It inserts itself into the membrane, where it forms conductive pores. Depending on its concentration, it eventually disintegrates the membrane in a detergent-like manner. The molecular details of this activity are not yet sufficiently understood, nor are the mechanisms that the surfactin producer employs to resist its own toxic product. We have previously shown that B. subtilis modifies its membrane lipid composition upon the onset of surfactin production, mainly increasing the cardiolipin content. Here we show that the increased cardiolipin content leads to a decreased surfactin-induced leakage of liposomes reconstituted from lipids isolated from the surfactin producer. This stabilizing effect of cardiolipin is concentration-dependent. Using a propidium iodide-based cell permeabilization assay, we further confirmed that the cytoplasmic membrane of the mutant B. subtilis strain lacking cardiolipin was substantially more susceptible to the action of surfactin, even though the amount of bound surfactin was the same as in the wild-type strain. We propose that membrane remodelling; due to the increase in cardiolipin content, contributes to the surfactin tolerance of B. subtilis.

Reduced cardiolipin content decreases respiratory chain capacities and increases ATP synthesis yield in the human HepaRG cells

Cardiolipin (CL) is a unique mitochondrial phospholipid potentially affecting many aspects of mitochondrial function/processes, i.e. energy production through oxidative phosphorylation. Most data focusing on implication of CL content and mitochondrial bioenergetics were performed in yeast or in cellular models of Barth syndrome. Previous work reported that increase in CL content leads to decrease in liver mitochondrial ATP synthesis yield. Therefore the aim of this study was to determine the effects of moderate decrease in CL content on mitochondrial bioenergetics in human hepatocytes. For this purpose, we generated a cardiolipin synthase knockdown (shCLS) in HepaRG hepatoma cells showing bioenergetics features similar to primary human hepatocytes. shCLS cells exhibited a 55% reduction in CLS gene and a 40% decrease in protein expression resulting in a 45% lower content in CL compared to control (shCTL) cells. Oxygen consumption was significantly reduced in shCLS cells compared to shCTL regardless of substrate used and energy state analyzed. Mitochondrial low molecular weight supercomplex content was higher in shCLS cells (+60%) compared to shCTL. Significant fragmentation of the mitochondrial network was observed in shCLS cells compared to shCTL cells. Surprisingly, mitochondrial ATP synthesis was unchanged in shCLS compared to shCTL cells but exhibited a higher ATP:O ratio (+46%) in shCLS cells. Our results suggest that lowered respiratory chain activity induced by moderate reduction in CL content may be due to both destabilization of supercomplexes and mitochondrial network fragmentation. In addition, CL content may regulate mitochondrial ATP synthesis yield.

Compensatory role of the cis‐trans‐isomerase and cardiolipin synthase in the membrane fluidity of Pseudomonas putida DOT‐T1E

In Gram-negative bacteria, cell membrane fluidity is influenced by phospholipid head group composition and linked fatty acids. Exposure of Pseudomonas putida to stressing agents results in short- and long-term modifications in membrane lipids. The main adaptive change observed in response to organic solvents in the short term is the cis- to trans-isomerization of unsaturated fatty acids in a reaction mediated by cis/trans-isomerase (CTI); whereas in the long term an increase in cardiolipin content takes place. Despite the interest of these genes in the context of stress responses, the transcriptional regulation of the cti and cls genes has not been studied before. The cti and cls (cardiolipin synthase) genes in the solvent-tolerant P. putida DOT-T1E strain form monocistronic units and are expressed from sigma-70 promoters. Expression from the cls promoter is sixfold higher in the stationary phase than in the log phase, and expression of the cls gene is not influenced by solvents. The cti gene is expressed at fairly constant levels in the log and stationary phase, but its level of expression is moderately upregulated in response to toluene. We used fluorescence polarization assays to show that mutants deficient in the cti gene exhibit less rigid membranes than the wild-type strain, whereas mutants with a knockout in the cls gene exhibit increased membrane rigidity. A double cti/cls mutant has similar membrane rigidity as the wild-type strain, which points towards a compensatory effect of the mutations with regard to membrane fluidity. However, the cls and cls/cti mutants were more sensitive to solvents than the wild-type and the cti mutant because of the impaired functioning of efflux drug transporters.

Membrane phospholipid synthesis in Escherichia coli: alteration by glycerol and physiological consequences in a pss mutant.

Escherichia coli mutants harboring the pss-1 allele (coding for a temperature-sensitive phosphatidylserine synthase) are temperature sensitive for growth and synthesize less phosphatidylethanolamine at higher temperatures, giving rise to abnormal membrane phospholipid compositions. To obtain information concerning the determinant for the phospholipid polar headgroup composition and the lethal factor in the defective membranes, we have examined the effect of increased supply of sn-glycerol 3-phosphate on the phospholipid synthesis and the growth ability of a pss-1 mutant. For this purpose, a pair of E. coli K-12 derivatives isogenic except for the pss-1 allele was constructed from strain BB26-36 to harbor the mutations related to glycerol metabolism (glpD3, glpR2, glpKi, and phoA8). Pulse- and uniform-labeling of phospholipids with 32P at 42 degrees C in a synthetic medium with (0.2%) or without glycerol showed that glycerol further lowered the temperature-sensitive formation of phosphatidylethanolamine, removed the phosphatidate and CDP-diacylglycerol accumulated in the absence of glycerol, and resulted in an increase in cardiolipin content in the pss-1 mutant. The phospholipid synthesis and contents in the pss+ strain were not significantly affected by glycerol. Glycerol in the medium markedly enhanced the growth defect of the pss-1 mutant, which was remediable by sucrose. The results indicate that the intracellular pool of sn-glycerol 3-phosphate is the limiting factor for acidic phospholipid synthesis in the pss-1 mutant, and cardiolipin unusually accumulated is injurious to the functional E. coli membranes. Possible determinants for the phospholipid composition of the wild-type E. coli cells are also discussed on the basis of the present observations.

Phospholipids and glycerides composition during spheroplasts formation of Mycobacterium smegmatis ATCC 14468.

The phospholipids and glycerides composition of spheroplasts of Mycobacterium smegmatis ATCC 14468 was examined. The percent total phospholipids in total lipids as well as cardiolipin were found to be higher in spheroplasts as compared to their parent forms. Increase in cardiolipin and free fatty acids content and decrease in triglycerides levels were observed during spheroplasts formation. The results suggest that increase in cardiolipin content in spheroplasts is an adaptational change concomitant with the loss of cell walls.

Alteration in phospholipid composition of Staphylococcus aureus during formation of autoplast

The phospholipid composition of autoplasts (protoplasts made by autolysis) of Staphylococcus aureus 209P was examined. The autoplasts were prepared by incubation of 209P cells in 1.2 M sucrose — 0.33 M acetate buffer (pH 5.8). Cardiolipin comprised nearly half the total phospholipid in these autoplasts. Autoplasts had a lower phosphatidylglycerol content than intact cells but a similar lysylphosphatidylglycerol content. The increase in cardiolipin content during release of autoplasts was not affected by pH or temperature. The result indicates that removal of the cell wall caused the increase in cardiolipin content. The total amount of phospholipids increased slightly during autoplast formation, but there was no significant increase in fatty acids or diglycerides. The changes of phospholipid composition during formation of the autoplasts was due to de novo synthesis of cardiolipin from phosphatidylglycerol.

Lipid-protein interactions in Escherichia coli. Membrane-associated f1 bacteriophage coat protein and phospholipid metabolism

The effects of insertion of the major coat protein of f1 bacteriophage into Escherichia coli membranes were investigated. The relative level of phosphatidylethanolamine decreased due to the failure to accumulate phosphatidylethanolamine when the cellular levels of phosphatidylglycerol and cardiolipin were increasing. This decreased accumulation was correlated with a 4-fold reduction in phosphatidylethanolamine synthesis. A 10- to 20-fold increase in cardiolipin content resulted from both a 3-fold increase in cardiolipin synthesis and a decrease in cardiolipin turnover. As long as cell division and protein synthesis continued, the number of cardiolipin molecules per coat protein molecules in the bacterium attained a constant value. The coat protein had little effect of phosphatidylglycerol synthesis. This data suggests that the coat protein froms a specific association with cardiolipin in the host membranes. Additional evidence suggests that cardiolipin also may facilitate the entry of coat protein into membranes.