Plasmalogens (vinyl-ether phospholipids) are an emergent class of lipid drugs against various diseases involving neuro-inflammation, oxidative stress, mitochondrial dysfunction, and altered lipid metabolism. They can activate neurotrophic and neuroprotective signaling pathways but low bioavailabilities limit their efficiency in curing neurodegeneration. Here, liquid crystalline lipid nanoparticles (LNPs) are created for the protection and non-invasive intranasal delivery of purified scallop-derived plasmalogens. The in vivo results with a transgenic mouse Parkinson's disease (PD) model (characterized by motor impairments and α-synuclein deposition) demonstrate the crucial importance of LNP composition, which determines the self-assembled nanostructure type. Vesicle and hexosome nanostructures (characterized by small-angle X-ray scattering) display different efficacy of the nanomedicine-mediated recovery of motor function, lipid balance, and transcriptional regulation (e.g., reduced neuro-inflammation and PD pathogenic gene expression). Intranasal vesicular and hexosomal plasmalogen-based LNP treatment leads to improvement of the behavioral PD symptoms and downregulation of the Il6, Il33, and Tnfa genes. Moreover, RNA-sequencing and lipidomic analyses establish a dramatic effect of hexosomal nanomedicines on PD amelioration, lipid metabolism, and the type and number of responsive transcripts that may be implicated in neuroregeneration.
- Keywords
- RNA‐seq, hexosomes, in vivo Parkinson's disease transgenic mouse model, liquid crystalline lipid nanoparticles, nanomedicine,
- MeSH
- Administration, Intranasal * MeSH
- Liposomes MeSH
- Lipid Metabolism drug effects MeSH
- Disease Models, Animal * MeSH
- Mice, Transgenic MeSH
- Mice MeSH
- Nanoparticles * chemistry MeSH
- Nanomedicine * methods MeSH
- Parkinson Disease * metabolism drug therapy MeSH
- Plasmalogens * chemistry pharmacology MeSH
- Gene Expression Regulation drug effects MeSH
- Animals MeSH
- Check Tag
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Lipid Nanoparticles MeSH Browser
- Liposomes MeSH
- Plasmalogens * MeSH
Plasmalogens are a group of lipids mainly found in the cell membranes. They occur in anaerobic bacteria and in some protozoa, invertebrates and vertebrates, including humans. Their occurrence in plants and fungi is controversial. They can protect cells from damage by reactive oxygen species, protect other phospholipids or lipoprotein particles against oxidative stress, and have been implicated as signaling molecules and modulators of membrane dynamics. Biosynthesis in anaerobic and aerobic organisms occurs by different pathways, and the main biosynthetic pathway in anaerobic bacteria was clarified only this year (2021). Many different analytical techniques have been used for plasmalogen analysis, some of which are detailed below. These can be divided into two groups: shotgun lipidomics, or electrospray ionization mass spectrometry in combination with high performance liquid chromatography (LC-MS). The advantages and limitations of both techniques are discussed here, using examples from anaerobic bacteria to specialized mammalian (human) organs.
- MeSH
- Bacteria, Anaerobic * MeSH
- Spectrometry, Mass, Electrospray Ionization MeSH
- Humans MeSH
- Lipidomics MeSH
- Lipids MeSH
- Plasmalogens * MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Review MeSH
- Names of Substances
- Lipids MeSH
- Plasmalogens * MeSH
The main bottleneck in the return of industrial butanol production from renewable feedstock through acetone-butanol-ethanol (ABE) fermentation by clostridia, such as Clostridium beijerinckii, is the low final butanol concentration. The problem is caused by the high toxicity of butanol to the production cells, and therefore, understanding the mechanisms by which clostridia react to butanol shock is of key importance. Detailed analyses of transcriptome data that were obtained after butanol shock and their comparison with data from standard ABE fermentation have resulted in new findings, while confirmed expected population responses. Although butanol shock resulted in upregulation of heat shock protein genes, their regulation is different than was assumed based on standard ABE fermentation transcriptome data. While glucose uptake, glycolysis, and acidogenesis genes were downregulated after butanol shock, solventogenesis genes were upregulated. Cyclopropanation of fatty acids and formation of plasmalogens seem to be significant processes involved in cell membrane stabilization in the presence of butanol. Surprisingly, one of the three identified Agr quorum-sensing system genes was upregulated. Upregulation of several putative butanol efflux pumps was described after butanol addition and a large putative polyketide gene cluster was found, the transcription of which seemed to depend on the concentration of butanol.
- Keywords
- Clostridium beijerinckii, ABE fermentation, butanol shock, transcriptome analysis,
- MeSH
- Biological Transport genetics MeSH
- Bioreactors microbiology MeSH
- Cell Membrane metabolism MeSH
- Butanols toxicity MeSH
- Clostridium beijerinckii drug effects genetics metabolism MeSH
- Stress, Physiological genetics MeSH
- Glucose metabolism MeSH
- Glycolysis genetics physiology MeSH
- Fatty Acids metabolism MeSH
- Plasmalogens biosynthesis MeSH
- Heat-Shock Proteins metabolism MeSH
- Quorum Sensing genetics MeSH
- Gene Expression Profiling MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Butanols MeSH
- Glucose MeSH
- Fatty Acids MeSH
- Plasmalogens MeSH
- Heat-Shock Proteins MeSH
Electrospray ionization-tandem mass spectrometry (ESI-MS/MS) was used for characterizing intact plasmalogen phospholipid molecules in beer-spoilage bacteria. Identification of intact plasmalogens was carried out using collision-induced dissociation and the presence of suitable marker molecular species, both qualitative and quantitative, was determined in samples containing the anaerobic bacteria Megasphaera and Pectinatus. Using selected ion monitoring (SIM), this method had a limit of detection at 1 pg for the standard, i.e. 1-(1Z-octadecenyl)-2-oleoyl-sn-glycero-3-phosphoethanolamine and be linear in the range of four orders of magnitude from 2 pg to 20 ng. This technique was applied to intact plasmalogen extracts from the samples of contaminated and uncontaminated beer without derivatization and resulted in the identification of contamination of beer by Megasphaera and Pectinatus bacteria. The limit of detection was about 830 cells of anaerobic bacteria, i.e. bacteria containing natural cyclopropane plasmalogenes (c-p-19:0/15:0), which is the majority plasmalogen located in both Megasphaera and Pectinatus. The SIM ESI-MS method has been shown to be useful for the analysis of low concentration of plasmalogens in all biological samples, which were contaminated with anaerobic bacteria, e.g. juice, not only in beer. Significance and impact of the study: Electrospray ionization-tandem mass spectrometry (ESI-MS/MS) using collision-induced dissociation was used to characterize intact plasmalogen phospholipid molecules in beer-spoilage anaerobic bacteria Megasphaera and Pectinatus. Using selected ion monitoring (SIM), this method has a detection limit of 1 pg for the standard 1-(1Z-octadecenyl)-2-oleoyl-sn-glycero-3-phosphoethanolamine and is linear within four orders of magnitude (2 pg to 20 ng). The limit of detection was about 830 cells of bacteria containing natural cyclopropane plasmalogen (c-p-19:0/15:0). SIM ESI-MS method is useful for analyzing low concentrations of plasmalogens in biological samples contaminated with anaerobic bacteria, e.g. beer or juice.
- Keywords
- Megasphaera, Pectinatus, beer, contamination, electrospray tandem mass spectrometry, plasmalogens,
- MeSH
- Spectrometry, Mass, Electrospray Ionization MeSH
- Limit of Detection MeSH
- Megasphaera classification isolation & purification metabolism MeSH
- Pectinatus classification isolation & purification metabolism MeSH
- Beer microbiology MeSH
- Plasmalogens analysis MeSH
- Food Microbiology methods MeSH
- Tandem Mass Spectrometry MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Plasmalogens MeSH
Changes in membrane lipid composition of Clostridium pasteurianum NRRL B-598 were studied during butanol fermentation by lipidomic analysis, performed by high resolution electrospray ionization tandem mass spectrometry. The highest content of plasmalogen phospholipids correlated with the highest butanol productivity, which indicated a probable role of these compounds in the complex responses of cells toward butanol stress. A difference in the ratio of saturated to unsaturated fatty acids was found between the effect of butanol produced by the cells and butanol added to the medium. A decrease in the proportion of saturated fatty acids during conventional butanol production was observed while a rise in the content of these acids appeared when butanol was added to the culture. The largest change in total plasmalogen content was observed one hour after butanol addition i.e. at the 7th hour of cultivation. When butanol is produced by bacterial cells, then the cells are not subjected to severe stress and responded to it by relatively slowly changing the content of fatty acids and plasmalogens, while after a pulse addition of external butanol (to a final non-lethal concentration of 0.5 % v/v) the cells reacted relatively quickly (within a time span of tens of minutes) by increasing the total plasmalogen content.
- MeSH
- Biomass MeSH
- Clostridium drug effects growth & development metabolism MeSH
- Spectrometry, Mass, Electrospray Ionization MeSH
- Fatty Acids analysis MeSH
- Membrane Lipids chemistry MeSH
- 1-Butanol metabolism pharmacology MeSH
- Fatty Acids, Unsaturated analysis MeSH
- Plasmalogens analysis MeSH
- Batch Cell Culture Techniques MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Fatty Acids MeSH
- Membrane Lipids MeSH
- 1-Butanol MeSH
- Fatty Acids, Unsaturated MeSH
- Plasmalogens MeSH
High resolution electrospray ionization tandem mass spectrometry (HR-ESI-MS/MS) was used to analyze cardiolipins (Ptd2Gro) including their plasmalogen forms from three species of the anaerobic beer-spoilage bacterial genus Pectinatus. Cardiolipins including their plasmalogens were analyzed by HR-ESI-MS/MS on Orbitrap and almost 100 cardiolipins (i.e. tetra-acyl--Ptd2Gro, plasmenyl-tri-acyl--PlsPtd2Gro, and di-plasmenyl-di-acyl--Pls2Ptd2Gro) of three classes were identified. The structures of the molecular species that consist of various regioisomers and structurally similar compounds were revealed in detail. The high resolution mass spectrometry allowed the unambiguous structural assignment of Ptd2Gro, PlsPtd2Gro, and Pls2Ptd2Gro in the three species of Pectinatus, which contain predominantly odd numbered fatty acids.
- MeSH
- Spectrometry, Mass, Electrospray Ionization methods MeSH
- Cardiolipins analysis chemistry MeSH
- Food Contamination MeSH
- Molecular Structure MeSH
- Pectinatus chemistry MeSH
- Beer microbiology MeSH
- Plasmalogens analysis chemistry MeSH
- Food Microbiology MeSH
- Tandem Mass Spectrometry methods MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Cardiolipins MeSH
- Plasmalogens MeSH
Lipids form a significant part of animal organs and they are responsible for important biological functions, such as semi-permeability and fluidity of membranes, signaling activity, anti-inflammatory processes, etc. We have performed a comprehensive nontargeted lipidomic characterization of porcine brain, heart, kidney, liver, lung, spinal cord, spleen, and stomach using hydrophilic interaction liquid chromatography (HILIC) coupled to electrospray ionization mass spectrometry (ESI/MS) to describe the representation of individual lipid classes in these organs. Detailed information on identified lipid species inside classes are obtained based on relative abundances of deprotonated molecules [M-H](-) in the negative-ion ESI mass spectra, which provides important knowledge on phosphatidylethanolamines and their different forms of fatty acyl linkage (ethers and plasmalogens), phosphatidylinositols, and hexosylceramides containing nonhydroxy- and hydroxy-fatty acyls. The detailed analysis of identified lipid classes using reversed-phase liquid chromatography in the second dimension was performed for porcine brain to determine more than 160 individual lipid species containing attached fatty acyls of different acyl chain length, double-bond number, and positions on the glycerol skeleton. The fatty acid composition of porcine organs is determined by gas chromatography with flame ionization detection after the transesterification with sodium methoxide.
- MeSH
- Chromatography, Liquid methods MeSH
- Chromatography, Gas methods MeSH
- Phosphatidylethanolamines analysis MeSH
- Phospholipids analysis MeSH
- Spectrometry, Mass, Electrospray Ionization methods MeSH
- Hydrophobic and Hydrophilic Interactions MeSH
- Liver chemistry MeSH
- Kidney chemistry MeSH
- Lipids analysis chemistry MeSH
- Fatty Acids analysis MeSH
- Spinal Cord chemistry MeSH
- Brain Chemistry MeSH
- Myocardium chemistry MeSH
- Plasmalogens analysis MeSH
- Lung chemistry MeSH
- Swine MeSH
- Spleen chemistry MeSH
- Stomach chemistry MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Phosphatidylethanolamines MeSH
- Phospholipids MeSH
- Lipids MeSH
- Fatty Acids MeSH
- Plasmalogens MeSH
Plasmalogens are a group of lipids with potentially important, and not yet fully known, functions in organisms from bacteria to protozoans, invertebrates, and mammals. They can protect cells against the damaging effects of reactive oxygen species, protect other phospholipids or lipoprotein particles against oxidative stress, and have been implicated as signaling molecules and modulators of membrane dynamics. They have been found in many anaerobic bacterial species, and their biosynthetic pathways differ in aerobic and anaerobic organisms. The use of advanced techniques permits the identification of not only plasmalogen classes but also their positional isomers and often also individual molecular species. This paper describes direct analyses of plasmalogens from natural sources, frequently very unusual, using electrospray ionization mass spectrometry in combination with high-performance liquid chromatography and/or shotgun lipidomics.
- MeSH
- Bacteria chemistry metabolism MeSH
- Molecular Structure MeSH
- Molecular Weight MeSH
- Plasmalogens chemistry metabolism MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Review MeSH
- Names of Substances
- Plasmalogens MeSH
Liquid chromatography-electrospray tandem mass spectrometry (LC/ESI-MS/MS) was used to analyze phospholipids from three species of the anaerobic beer-spoilage bacterial genus Pectinatus. Analysis of total lipids by HILIC (Hydrophilic Interaction Liquid Chromatography) column succeeded in separating diacyl- and plasmalogen phospholipids. Plasmalogens were then analyzed by means of the ESI-MS/MS and more than 220 molecular species of four classes of plasmalogens (PlsCho (choline plasmalogen), PlsEtn (ethanolamine plasmalogen), PlsGro (glycerol plasmalogen), and PlsSer (serine plasmalogen)) were identified. Major molecular species were c-p19:0/15:0 PlsEtn and PlsSer, which accounted for more than 4% of the total lipids.
- MeSH
- Aldehydes chemistry MeSH
- Phospholipids analysis MeSH
- Spectrometry, Mass, Electrospray Ionization methods MeSH
- Humans MeSH
- Fatty Acids chemistry MeSH
- Molecular Structure MeSH
- Pectinatus chemistry MeSH
- Beer microbiology MeSH
- Plasmalogens analysis MeSH
- Tandem Mass Spectrometry methods MeSH
- Chromatography, High Pressure Liquid methods MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Aldehydes MeSH
- Phospholipids MeSH
- Fatty Acids MeSH
- Plasmalogens MeSH
The aim of this study was to determine the concentration of phospholipids (PL), plasmalogen components of choline (PC) and ethanolamine (PE) phosphoglycerides (PLPC, PLPE) and fatty acid profile of PL and triacylglycerols (TAG) in developing rat left ventricular myocardium between postnatal day (d) 2 and 100. The steepest increase of total PL (TPL) concentration occurs between d2 and d5, followed by a further slower increase between d20 and d40. Similar developmental changes were observed in PC and PE. The PLPE concentration rises by d10, whereas PLPC does not change during the whole period investigated, except for the transient decline on d5. The concentration of diphosphatidylglycerol (DPG) increases by d60; the steepest rise occurs between d20 and d40. Phosphatidylinositol (PI) concentration rises only by d5. The concentration of phosphatidylserine (PS) decreases between d5 and d10 and then it does not change. Sphingomyelin (SM) concentration is maintained till d10, it declines on d20 and does not change thereafter. The proportion of saturated fatty acids (SFA) increases by d5 in PC, PE, PS and TAG, and by d10 in DPG and PI. After d20 the SFA proportion gradually decline in all lipids. Monounsaturated FA (MUFA) proportion decreases in PC, PE, PI and PS from d2 till d10, and in the weaning period it tends to rise again. In contrast, in DPG and TAG the proportion of MUFA declines during the whole postnatal period. N-6 polyunsaturated FA (PUFA) decrease in all PL by d20 and rise again thereafter; in TAG they decline between d2 and d10 and return to the initial level by d100. N-3 PUFA increase in all PL during the suckling period and decline after weaning; in TAG they increase only by d5 and then they decline. This remodeling of myocardial PL and TAG composition during postnatal development may affect membrane properties and contribute to developmental changes in the function of membrane proteins and cell signaling.
- MeSH
- Choline metabolism MeSH
- Phosphatidylinositols analysis metabolism MeSH
- Phosphatidylserines analysis metabolism MeSH
- Phospholipids chemistry metabolism MeSH
- Rats MeSH
- Fatty Acids analysis metabolism MeSH
- Myocardium chemistry metabolism MeSH
- Plasmalogens metabolism MeSH
- Rats, Wistar MeSH
- Heart growth & development MeSH
- Heart Ventricles cytology MeSH
- Triglycerides metabolism MeSH
- Animals MeSH
- Check Tag
- Rats MeSH
- Male MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Choline MeSH
- Phosphatidylinositols MeSH
- Phosphatidylserines MeSH
- Phospholipids MeSH
- Fatty Acids MeSH
- Plasmalogens MeSH
- Triglycerides MeSH
