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Glucosylceramides

Glucosylceramides are a class of glycosphingolipids found in the plasma membrane of many cell types.
They play a crucial role in cellular signaling, membrane structure, and lipid raft formation.
Resaerch into Glucosylceramides has implications for understanding and treating various disosders, including Gaucher disease, a rare lysosomal storage disorder.
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Most cited protocols related to «Glucosylceramides»

Quantification of Endogenous Lipids in Mouse Tissues

Endogenous lipids from mouse liver and heart were detected and quantified using several techniques. FC was quantified using straight-phase HPLC and ELS detection as previously described10 (link). Quantification was made against an external calibration curve. This chromatographic set-up was also used to fractionate DG. Quantification of CE, TG, SM, and phospholipids (all from the total extract) and DG (fractionated from the HPLC) was made by direct infusion (shotgun) on a QTRAP 5500 mass spectrometer (Sciex, Concord, Canada) equipped with a robotic nanoflow ion source, TriVersa NanoMate (Advion BioSciences, Ithaca, NJ)11 (link). For this analysis, total lipid extracts, stored in chloroform:methanol (2:1), were diluted with internal standard-containing chloroform/methanol (1:2) with 5mM ammonium acetate and then infused directly into the mass spectrometer. The characteristic dehydrocholesterol fragment m/z 369.3 was selected for precursor ion scanning of CE in positive ion mode12 (link). The analysis of TG and DG was performed in positive ion mode by neutral loss detection of 10 common acyl fragments formed during collision induced dissociation13 (link). The PC, LPC and SM were detected using precursor ion scanning of m/z 184.114 (link), while the PE, phosphatidylserine (PS), phosphatidylglycerol (PG) and phosphatidylinositol (PI) lipid classes were detected using neutral loss of m/z 141.0, m/z 185.0, m/z 189.0 and m/z 277.0 respectively15 (link)16 (link). For quantification, lipid class-specific internal standards were used. The internal standards were either deuterated or contained diheptadecanoyl (C17:0) fatty acids.
Ceramides (CER), dihydroceramides (DiCER), glucosylceramides (GlcCER) and lactosylceramides (LacCER) were quantified using a QTRAP 5500 mass spectrometer equipped with a Rheos Allegro quaternary ultra-performance pump (Flux Instruments, Basel, Switzerland). Before analysis the total extract was exposed to alkaline hydrolysis (0.1M potassium hydroxide in methanol) to remove phospholipids that could potentially cause ion suppression effects. After hydrolysis the samples were reconstituted in chloroform:methanol:water [3:6:2] and analyzed as previously described17 (link).
For the recovery experiments the tissue samples were spiked with non-endogenously present lipids (or endogenous lipids spiked at relatively high levels) and could therefore all be detected by lipid class specific scans using the shotgun approach. In the recovery experiment we therefore also included the PA and phosphatidylcholine plasmalogen (PC P) lipid class, which we could not measure endogenously using our current analytical platform. Due to poor ionization efficiency, FC was derivatized and analyzed as picolinyl esters according to previous publication18 (link). See Table 1 for details. With some exceptions, lipids are annotated according to Liebisch et al.19 (link).

Löfgren L., Forsberg G.B, & Ståhlman M. (2016). The BUME method: a new rapid and simple chloroform-free method for total lipid extraction of animal tissue. Scientific Reports, 6, 27688.

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Publication 2016

Allegro ammonium acetate Ceramides Chloroform Chromatography Dehydrocholesterols dihydroceramide Esters Fatty Acids Glucosylceramides Heart High-Performance Liquid Chromatographies Hydrolysis Lactosylceramides Lipids Liver Methanol Mice, House Phosphatidylcholines Phosphatidyl Glycerol Phosphatidylinositols Phosphatidylserines Phospholipids Plasmalogens potassium hydroxide Radionuclide Imaging Tissues

Lipid Extraction and Mass Spectrometry Analysis

Total lipids were extracted as described in (Singh et al., 2011 ). In brief, single colony from WT, Δsmt1, Δsmt1+SMT1, Δgcs1 and Δgcs1+GCS1 were picked from freshly streaked YPD plates and grown in 15 ml YPD at 30°C for 20 hrs at 250 rpm. The cells were washed twice with sterile Phosphate Buffered Saline (PBS) and counted. Then, 5 × 10⁸ cells were placed in a single glass tube, washed once with sterile DDW, and Mandala extraction was performed for extraction of inositol-containing phospholipids and phosphatidylcholine. Bligh and Dyer lipid extraction (Bligh et al., 1959 ) of neutral lipids was performed on the dried lipids obtained after Mandala lipid extraction (Mandala et al., 1995 (link)). A quarter of the samples were aliquoted for inorganic phosphate determination. The tubes were vacuum dried and used for mass spectrometry (MS) analysis. The MS and MS/MS scans were conducted using a Thermo Finnigan TSQ7000 triple quadruple mass spectrometer with electrospray ionization (Bielawski et al., 2009 (link)). The analyses included multiple reaction monitoring (MRM) for the characteristic product ions of m/z=276.2 (α-OH-Δ4-Δ8,9methyl-GlcCer at m/z 756.4), 262.0 (α-OH-Δ4-Δ8- GlcCer at m/z 742.4), 576.4 (α-OH-Δ4-Δ8,9methyl-ceramide at m/z 594.4), and 562.4 (α-OH-Δ4-Δ8-ceramide at m/z 580.4).

Singh A., Wang H., Silva L.C., Na C., Prieto M., Futerman A.H., Luberto C, & Del Poeta M. (2012). Methylation of glycosylated sphingolipid modulates membrane lipid topography and pathogenicity of Cryptococcus neoformans. Cellular Microbiology, 14(4), 500-516.

Publication 2011

Cells Ceramides Faciothoracoskeletal Syndrome Glucosylceramides Ions Lipids Mass Spectrometry Phosphates Phosphatidylcholines Phosphatidylinositols Radionuclide Imaging Saline Solution Sterility, Reproductive Tandem Mass Spectrometry Vacuum

Sphingolipid Analysis by Mass Spectrometry

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Publication 2011

Acids Biopharmaceuticals Complex Mixtures Fatty Acids Gangliosides Glucosylceramides Head Hydroxy Acids Hypersensitivity M-200 Phosphorylcholine phytosphingosine Radionuclide Imaging Retention (Psychology) sphinganine Sphingolipids Sphingosine Tandem Mass Spectrometry Vertebral Column

Comprehensive Lipidomic and Metabolomic Analysis of Brain Tissue

During the sample preparation, lipids and polar metabolites were separated prior to analyses through solvent extraction/fractionation. Hence, potential problems in ion suppression or the ability of compounds to be ionized were limited due to the use of Lipidomic LC–MS/MS, HILIC-MS/MS (including positive and negative electrospray), and the complementary use of GC-electron ionization-MS.
Briefly, five milligrams of tissue from each brain region were homogenized in 225 µL of −20 °C cold, internal standard-containing methanol using a GenoGrinder 2010 (SPEX SamplePrep) for 2 min at 1,350 rpm. The extraction methanol contained the following internal standards for quality control and retention time normalization: sphingosine (d17:1), LPE (17:1), LPC (17:0), MG (17:0/0:0/0:0), DG (12:0/12:0/0:0), PC (12:0/13:0), cholesterol-d₇, SM (18:1/17:1), ceramide (d18:1/17:0), PE (17:0/17:0), TG (14:0/16:1/14:0)-d₅, TG (17:0/17:1/17:0)-d₅, acylcarnitine (18:1)-d₃, fatty acid (16:0)-d₃, MAG (17:0/0:0/0:0), PI (15:0–18:1)-d₇, PG (17:0/17:0), PS (15:0-18:1)-d₇, glucosylceramide(d18:1/17:0), mono-sulfo galactosylceramide(d18:1/17:0), and 5-PAHSA-d₉. The homogenate was vortexed for 10 s. 750 µL of −20 °C cold, internal standard-containing methyl tertiary-butyl ether (MTBE) was added, and the mixture was vortexed for 10 s and shaken at 4 °C for 5 min with an Orbital Mixing Chilling/Heating Plate (Torrey Pines Scientific Instruments). MTBE contained cholesteryl ester 22:1 as internal standard. Next, 188 µL room temperature water was added and vortexed for 20 s to induce phase separation. After centrifugation for 2 min at 14,000×g, two 350 µL aliquots of the upper non-polar phase and two 125 µL aliquots of the bottom polar phase were collected and dried down. The remaining fractions were combined to form QC pools and were injected after every set of 10 biological samples.
The non-polar phase employed for lipidomics was resuspended in a mixture of methanol/toluene (60 µL, 9:1, v/v) containing an internal standard [12-[(cyclohexylamine) carbonyl]amino]-dodecanoic acid (CUDA)] before injection. Resuspension of dried polar phases for HILIC analysis was performed in a mixture of acetonitrile/water (90 µL, 4:1, v/v) containing the following internal standards: CUDA, caffeine-d₉, acetylcholine-d₄, TMAO-d₉, 1-methylnicotinamide-d₃, Val-Tyr-Val, betaine-d₉, acyl carnitine (2:0)-d₃, N-methyl-histamine-d₃, l-carnitine-d₃, butyrobetaine-d₉, l-glutamine-d₅, aspartic acid-d₃, l-arginine-¹⁵N₂, cystine-d₄, asparagine-d₃, histidine-d₅, isoleucine-d₁₀, leucine-d₁₀, methionine-d₈, ornithine-d₂, phenylalanine-d₈, proline-d₇, threonine-d₅, tryptohan-d₈, tyrosine-d₇, valine-d₈, spermine-d₈, glucose-d₇, fructose-6-phosphate-¹³C₆, succinic acid-d₄, taurocholic acid-d₄, adenosine 5′-monophosphate-¹⁵N₅, uridine 5′-monophosphate-¹⁵N₂, dopamine-d₄, taurine-d₄, uracil-d₂, biotin-d₄, N-acetylalanine-d₃, guanine-¹³C, and adenosine-¹³C₅. The second dried polar phase was reserved for GC analysis and a following derivatization process was carried out before injection. First, carbonyl groups were protected by methoximation with methoxyamine hydrochloride in pyridine (40 mg/mL, 10 µL) was added to the dried samples. Then, the mixture was incubated at 30 °C for 90 min followed by trimethylsilylation with N-methyl-N-(trimethylsilyl) trifluoroacetamide (MSTFA, 90 μL) containing C8–C30 fatty acid methyl esters (FAMEs) as internal standards by shaking at 37 °C for 30 min.

Ding J., Ji J., Rabow Z., Shen T., Folz J., Brydges C.R., Fan S., Lu X., Mehta S., Showalter M.R., Zhang Y., Araiza R., Bower L.R., Lloyd K.C, & Fiehn O. (2021). A metabolome atlas of the aging mouse brain. Nature Communications, 12, 6021.

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Publication 2021

5-PAHSA acetonitrile Acetylcholine acylcarnitine Adenosine Adenosine Monophosphate Amino Acids Arginine Asparagine Aspartic Acid Betaine Biopharmaceuticals Biotin Brain Caffeine Centrifugation Ceramides Cholesterol Cholesterol Esters Cold Temperature Cyclohexylamines Cystine Dopamine Electrons Esters Ethyl Ether Fatty Acids Fractionation, Chemical fructose-6-phosphate Galactosylceramides gamma-butyrobetaine Gas Chromatography-Mass Spectrometry Glucose Glucosylceramides Glutamine Guanine Histidine Isoleucine Leucine Levocarnitine Lipids Methanol Methionine methoxyamine methoxyamine hydrochloride Methylhistamines N(1)-methylnicotinamide N-acetylalanine N-methyl-N-(trimethylsilyl)trifluoroacetamide Ornithine PC 12 ester Phenylalanine Pinus Proline PS 15 pyridine pyridine hydrochloride Retention (Psychology) Solvents Spermine Sphingosine Succinic Acid Tandem Mass Spectrometry Taurine Taurocholic Acid Threonine Tissues Toluene trifluoroacetamide trimethyloxamine Tyrosine Uracil Uridine Monophosphate Valine valylvaline

Comprehensive Metabolomics Profiling Analysis

In total, 363 different metabolites were detected. The metabolomics dataset contains 18 amino acids, nine reducing mono-, di- and oligosaccharides (abbreviated as Hn for n-hexose, dH for desoxyhexose, UA for uronic acid, HNAc for N-acetylglucosamine, P for Pentose, NANA for N-acetylneuraminic-acid), seven biogenic amines, five prostaglandins, arachidonic acid (AA), docosahexaenoic acid (DHA), free carnitine (C0), 28 acylcarnitines (Cx:y), hydroxylacylcarnitines (C(OH)x:y), and dicarboxylacylcarnitines (Cx:y-DC), 85 ceramides (Cer), glucosylceramides (GlcCer), different sphingomyelins (SMx:y) and sphingomyelin-derivatives, such as N-hydroxyldicarboacyloylsphingosyl-phosphocholine (SM(OH,COOH)x:y) and N-hydroxylacyloylsphingosyl-phosphocholine (SM(OH)x:y). In addition, 208 phospholipids were detected, including different glycero-phosphatidic acids (PA), glycero-phosphatidylcholines (PC), glycero-phosphatidylethanolamines (PE), phosphatidylglycerols (PG), glycero-phosphatidylinositols (PI), glycero-phosphatidylinositol-bisphosphates (PIP2), and glycero-phosphatidylserines (PS). Glycerophospholipids are further differentiated with respect to the presence of ester (a) and ether (e) bonds in the glycerol moiety, where two letters (aa = diacyl, ae = acyl-alkyl, ee = dialkyl) denote that two glycerol positions are bound to a fatty acid residue, while a single letter (a = acyl or e = alkyl) indicates the presence of a single fatty acid residue. Lipid side chain composition is abbreviated as Cx:y, where x denotes the number of carbons in the side chain and y the number of double bonds. E.g. “PC ae C33:1” denotes a plasmalogen/plasmenogen phosphatidylcholine with 33 carbons in the two fatty acid side chains and a single double bond in one of them. The precise position of the double bonds and the distribution of the carbon atoms in different fatty acid side chains cannot be determined with this technology. In some cases, the mapping of metabolite names to individual masses can be ambiguous. For example, stereo-chemical differences are not always discernible, neither are isobaric fragments. In such cases, possible alternative assignments are indicated.

Gieger C., Geistlinger L., Altmaier E., Hrabé de Angelis M., Kronenberg F., Meitinger T., Mewes H.W., Wichmann H.E., Weinberger K.M., Adamski J., Illig T, & Suhre K. (2008). Genetics Meets Metabolomics: A Genome-Wide Association Study of Metabolite Profiles in Human Serum. PLoS Genetics, 4(11), e1000282.

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Publication 2008

Acetylglucosamine acylcarnitine Amino Acids Arachidonic Acid Biogenic Amines Carbon Carnitine Ceramides derivatives Docosahexaenoic Acids Esters Ethers Fatty Acids Glucosylceramides Glycerin Glycerophospholipids Hexoses Lipids N-Acetylneuraminic Acid Oligosaccharides Pentoses Phosphatidic Acids Phosphatidylcholines Phosphatidylethanolamines Phosphatidylglycerols Phosphatidylinositols Phosphatidylserines Phospholipids Phosphorylcholine Plasmalogens Prostaglandins single bond Sphingomyelins Uronic Acids

Most recents protocols related to «Glucosylceramides»

Immunofluorescence Analysis of Lysosomal and Autophagic Markers in hDF Cells

hDF cells (20,000 cells in 24-well plates on coverslip) were grown in complete medium for 24 h. Starvation was performed for 1 h in Hanks’ Balanced Salt solution (HBSS) before fixing with paraformaldehyde (PFA) 4% and staining with anti-LAMP1, anti-LAMP2, anti-p62, anti-LC3B, anti-TMEM175, anti-GC, and anti-GM antibodies. Briefly, incubation with blocking buffer (1 × PBS/0.1% triton/5% serum) was performed for 15 min in a humidified chamber. Primary and then secondary antibody incubations (1 × PBS/0.1% triton/antibody) were performed at 4 °C overnight and at RT for 1 h, respectively. Anti-LAMP1 (ab25630, Abcam, Cambridge, UK, 1:20), anti-LAMP2 (ab25631, Abcam, Cambridge, UK, 1:100), anti-LC3B (2775, Cell Signaling, Danvers, MA, USA, 1:200), anti-TMEM175 (19,925–1-AP, Proteintech, Manchester, UK, 1:100) and anti-p62/SQSTM1 (P0067, Sigma-Aldrich, St. Louis, MO, USA, 1:500), anti-GlcCer (RAS0010, Glycobiotech, Kuekels, Germany, 1:50) primary antibodies and anti-mouse IgG-Cy3 (C2181, Sigma-Aldrich, St. Louis, MO, USA, 1:200), anti-rabbit IgG-Cy3 (C2306, Sigma-Aldrich, St. Louis, MO, USA, 1:200), Donkey anti-Mouse IgG (H + L), Alexa Fluor™ 488 (A-21202, Thermo Fisher Scientific CA USA, 1:400), and Donkey anti-Rabbit IgG (H + L) Alexa Fluor™ 488 (A-21206, Thermo Fisher Scientific CA USA, 1:400) secondary antibodies were used. GM1 was detected through 2-h incubation with Cholera toxin subunit B (CT-B) conjugated with Alexa fluor 488 (c34775, Molecular Probes, Eugene, OR, USA, 1:50). Hoechst H3570 (Thermo Fisher Scientific, CA, USA, 1:1000) was used to visualize the nuclei. Slides were visualized with Nikon Confocal Microscope A1R and with Nikon Eclipse Ni-E Microscope.

Palomba N.P., Fortunato G., Pepe G., Modugno N., Pietracupa S., Damiano I., Mascio G., Carrillo F., Di Giovannantonio L.G., Ianiro L., Martinello K., Volpato V., Desiato V., Acri R., Storto M., Nicoletti F., Webber C., Simeone A., Fucile S., Maglione V, & Esposito T. (2023). Common and Rare Variants in TMEM175 Gene Concur to the Pathogenesis of Parkinson’s Disease in Italian Patients. Molecular Neurobiology, 60(4), 2150-2173.

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Publication 2023

alexa fluor 488 Anti-Antibodies anti-IgG Antibodies Buffers Cell Nucleus Cells Choleragenoid Equus asinus Glucosylceramides Hanks Balanced Salt Solution Immunoglobulins LAMP2 protein, human lysosomal-associated membrane protein 1, human Microscopy Microscopy, Confocal Molecular Probes Mus paraform Rabbits Serum

Fluorescent Lipid Probes for Membrane Characterization

Miltefosine (#850337) and most NBD-lipids were purchased from Avanti Polar Lipids (Alabaster, AL, USA), including 1-palmitoyl-2-{6-[(7-nitro-2-1,3-benzoxadiazol-4-yl)amino]hexanoyl}-sn-glycero-3-phosphocholine (NBD-PC; #810130), 1-palmitoyl-2-{6-[(7-nitro-2-1,3-benzoxadiazol-4-yl)amino]hexanoyl}-sn-glycero-3-phosphoethanol-amine (NBD-PE; #810153), 1-palmitoyl-2-{6-[(7-nitro-2-1,3-benzoxadiazol-4-yl)amino]hexanoyl}-sn-glycero-3-phosphoserine (ammonium salt) (NBD-PS; #810192), 1-palmitoyl-2-{6-[(7-nitro-2-1,3-benzoxadiazol-4-yl)amino]hexanoyl}-sn-glycero-3-[phospho-rac-(1-glycerol)] (ammonium salt) (NBD-PG; #810163), N-[6-[(7-nitro-2-1,3-benzoxadiazol-4-yl)amino]hexanoyl]-sphingosine-1-phosphocholine (NBD-SM; #810218), N-[6-[(7-nitro-2-1,3-benzoxadiazol-4-yl)amino]hexanoyl]-D-galactosyl-β1-1′-sphingosine (NBD-GalCer; #810220), and N-[6-[(7-nitro-2-1,3-benzoxadiazol-4-yl)amino]hexanoyl]-D-glucosyl-β1-1′-sphingosine (NBD-GlcCer; #810222). N-[(1S,2R,3E)-1-[[(4-O-beta-D-galactopyranosyl-beta-D-glucopyranosyl)oxy]methyl]-2-hydroxy-3-heptadecen-1-yl]-hexadecanamide-d3 (NBD-LacCer, #Cay24625-1) was purchased from Biomol (Hamburg, Germany). The detergent n-dodecyl-β-D-maltopyranoside (DDM) was purchased from GlyconBiochemicals GmbH (Luckenwalde, Germany). Unless otherwise indicated, chemicals were obtained from Sigma-Aldrich (München, Germany). Protease inhibitor cocktail contained aprotinin (1 mg/mL), leupeptin (1 mg/mL), pepstatin A (1 mg/mL; Roth), antipain (5 mg/mL), and benzamidine (0.157 mg/mL) in dimethylsulfoxide and was used at a 1:1000 dilution.

Veit S., Laerbusch S., López-Marqués R.L, & Günther Pomorski T. (2023). Functional Analysis of the P-Type ATPases Apt2-4 from Cryptococcus neoformans by Heterologous Expression in Saccharomyces cerevisiae. Journal of Fungi, 9(2), 202.

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Publication 2023

Alabaster Antipain Aprotinin benzamidine Chloride, Ammonium Detergents Glucosylceramides Glycerin Glycerylphosphorylcholine hexadecanamide leupeptin Lipids miltefosine N-(7-nitro-2,1,3-benzoxadiazol-4-yl)phosphatidylserine N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)phosphatidylethanolamine NBD-galactosylceramide nitrobenzoxadiazolyl-conjugated phosphatidylcholine pepstatin phosphoethanolamine Phosphorylcholine Phosphoserine Protease Inhibitors Sphingosine sphingosyl beta-glucoside Sulfoxide, Dimethyl Technique, Dilution

Comprehensive Sphingolipid Analysis Protocol

FcR Blocking Reagent, Viobility™ 488/520 Fixable Dye, Ly-6G VioBlue^® (REAfinity™, clone REA526), CD45 VioGreen^® (REAfinity™, clone REA737), CD11c PE-Vio^® 770 (clone REA754), and F4/80 APC (clone REA126) were purchased from Miltenyi Biotec (Bergisch Gladbach, Germany). Zymosan was obtained from InvivoGen (Toulouse, France). All other analytes and reagents were purchased from Sharlab (Sharlab S.L., Sentmenat, Spain), or Sigma-Aldrich (Sigma-Aldrich Chemical Co., Saint Quentin Fallavier, France). All the reagents were HPLC analytical grade, except those used for UHPLC-MSMS, which were of LC-MS grade. The 10 sphingolipids used as internal standards obtained from Sigma-Aldrich were manufactured by Avanti Polar Lipids under the trade name “Ceramide/Sphingoid Internal Standard Mixture I” and corresponded to C17 sphingosine (d17:1), C17 sphinganine (d17:0), C17 sphingosine-1-P (d17:1P), C17 sphinganine-1-P (d17:0P), lactosyl (ß) C12 ceramide (Lac18:1/12:0), C12 sphingomyelin (SM18:1/12:0), glucosyl (ß) C12 ceramide (Glu18:1/12:0), 12:0 ceramide (18:1/12:0), 12:0 ceramide-1-P (18:1/12:0P), and 25:0 ceramide (18:1/25:0) in ethanol solution. The 33 sphingolipids used as standards obtained from Sigma-Aldrich were solubilized in ethanol at a concentration of 25 µM and corresponded to deoxysphingosine (dSo = m18:1); deoxysphinganine (dSa = m18:0); sphingosine (So = d18:1); sphinganine (Sa = d18:0); sphingosine-1-P (d18:1P); sphinganine-1-P (d18:0P); glucosylsphingosine (GluSo); lysosphingomyelin (LysoSM); lactosylsphingosine (LacSo); N-acetylsphingosine (18:1/2:0); N-acetylsphinganine (18:0/2:0); ceramides: 18:1/14:0, 18:1/16:0, 18:1/18:0, 18:1/20:0, 18:1/22:0, 18:1/24:1, and 18:1/24:0; ceramide-1P: 18:1/16:0P; dihydroceramides: 18:0/16:0, and 18:0/24:0; glucosylceramides: Glu18:1/16:0 and Glu18:1/24:1; lactosylceramides: Lac18:1/16:0 and Lac18:1/24:1; sphingomyelins: SM18:1/14:0, SM18:1/16:0, SM18:1/18:0, SM18:1/18:1, SM18:1/20:0, SM18:1/22:0, SM18:1/24:1, and SM18:1/24:0.

Pierron A., Guzylack-Piriou L., Tardieu D., Foucras G, & Guerre P. (2023). Zymosan-Induced Murine Peritonitis Is Associated with an Increased Sphingolipid Synthesis without Changing the Long to Very Long Chain Ceramide Ratio. International Journal of Molecular Sciences, 24(3), 2773.

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Publication 2023

Cardiac Arrest Ceramides Clone Cells dihydroceramide Ethanol Glucosylceramides High-Performance Liquid Chromatographies Lactosylceramides lactosylsphingosine Lipids N-acetylsphingosine sphinganine Sphingolipids Sphingomyelins Sphingosine sphingosine phosphorylcholine sphingosyl beta-glucoside Zymosan

Comprehensive Lipidomic Analysis Protocol

The following lipid names and abbreviations are used. Cer – Ceramide, Chol – Cholesterol, CL – cardiolipin, DAG – Diacylglycerol, HexCer – Glucosyl/Galactosyl Ceramide, PA – Phosphatidic Acid, PC – Phosphatidylcholine, PE – Phosphatidyl-ethanolamine, PG – Phosphatidyl-glycerol, PI – Phosphatidylinositol, PS – Phosphatidylserine, and their respective lysospecies: lysoPA, lysoPC, lysoPE, lysoPI and lysoPS; and their ether derivatives: PC O-, PE O-, LPC O-, LPE O-; SE – Sterol Ester, SM – Sphingomyelin, TAG – Triacylglycerol.
Lipid species were annotated according to their molecular composition as follows: [lipid class]-[sum of carbon atoms in the fatty acids]:[sum of double bonds in the fatty acids];[sum of hydroxyl groups in the long chain base and the fatty acid moiety] (e.g., SM-32:2;1). Where available, individual fatty acid composition following the same rules is given in brackets (e.g., 18:1;0-24:2;0).

Beltran L.C., Cvirkaite-Krupovic V., Miller J., Wang F., Kreutzberger M.A., Patkowski J.B., Costa T.R., Schouten S., Levental I., Conticello V.P., Egelman E.H, & Krupovic M. (2023). Archaeal DNA-import apparatus is homologous to bacterial conjugation machinery. Nature Communications, 14, 666.

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Publication 2023

Carbon Cardiolipins Ceramides Cholesterol derivatives Diacylglycerol Esters Ethers Fatty Acids Glucosylceramides Hydroxyl Radical Lipids LYSO-PC lysophosphatidylinositol Phosphatidic Acid Phosphatidylcholines phosphatidylethanolamine Phosphatidyl Glycerol Phosphatidylinositols Phosphatidylserines SM(32) Sphingomyelins Sterols Triglycerides

Comprehensive Lipid Profiling Protocol

Data for each timepoint was classified by LIPID MAPS category^{66 (link)}: glycerophospholipids, glycerolipids, sphingolipids, sterol lipids, fatty acyls, and prenol lipids were detected. Each category contains distinct subclasses as annotated below. Glycerophospholipids: Phosphatidylcholine (PC), Phosphatidylethanolamine (PE), Phosphatidylserine (PS), Phosphatidylinositol (PI), Methylphosphocholine (MePC), Phosphatidic acid (PA), Bis-methyl phosphatidic acid (BisMePA), Dimethyl phosphatidylethanolamine (dMePE), Phosphatidylgylcerol (PG), Bis-methylphosphatidylserine (BisMePS), Bis-methyl phosphatidyl ethanolamine (BisMePE), Cardiolipin (CL), Phosphatidylethanol (PEt), Biotinyl-phosphoethanolamine (BiotinylPE), Phosphatidylmethanol (PMe), Phosphatidylinositol-bisphosphate (PIP2), Phosphatidylinositol-monophosphate (PIP), Lysophosphatidylcholine (LPC), Lysophosphatidylethanolamine (LPE), Lysophosphatidylserine (LPS), Lysophosphatidylinositol (LPI), Lysophosphosphatidylgylcerol (LPG), Lysodimethyl phosphatidyl ethanolamine (LdMePE). Glycerolipids: Triglyceride (TG), Monogalactosyldiacylglycerol (MGDG), Monogalactosylmonoacylglycerol (MGMG), Diglyceride (DG), Sulfoquinovosylmonoacylglycerol (SQMG), Sulfoquinovosyldiacylglycerol (SQDG). Sphingolipids: Hexosylceramides (Hex1Cer), Simple Glc series (CerG1), Sphingomyelin (SM), Ceramide (Cer), Ceramide phosphate (CerP), Sulfatide (ST), Sphingoid base (So), Sphingomyelin phytosphingosine (phSM), Simple Glc series (CerG2GNAc1), Ceramide phosphorylethanolamine (CerPE), Sphingosine (SPH), Dihexosylceramides (Hex2Cer). Sterol lipids: Cholesterol ester (ChE), Zymosterol (ZyE). Fatty acyls: Fatty acid (FA), Acyl Carnitine (AcCa). Prenol lipids: Coenzyme (Co).
Individual lipid species were annotated according to sum composition of carbons and double bonds in the format Lipid Subclass (total number of carbons: total number of double bonds). If distinct fatty acid chains could be identified, they were annotated separated by an underscore (ex. PC 32:1, or PC (16:0_16:1). Using this approach, we cannot determine the sn-1 or sn-2 positions of the fatty acid chains. Lipid species within the sphingolipid category contain prefixes ‘d’ or ‘t’ to denote di-hydroxy or tri-hydroxy bases. For example, SM(d18:1_23:0) contains 2 hydroxyl groups. The Hex1Cer subclass is comprised of both glucosylceramide (GlcCer) and galactosylceramide (GalCer); the orientation of one of the hydroxyl groups in Glc differs from in Gal, and thus cannot be resolved by these methods ^{67 (link)}. Plasmanyl lipid species (ether linked) are annotated by ‘e’ and plasmenyl/plasmalogen (vinyl ether linked) lipid species are annotated by ‘p’ (ex. PC (36:5e) or PE (16:0p_20:4) ^{68 (link)}.

Shing K., Sapp E., Boudi A., Liu S., Seeley C., Marchionini D., DiFiglia M, & Kegel-Gleason K.B. (2023). Early whole-body mutant huntingtin lowering averts changes in proteins and lipids important for synapse function and white matter maintenance in the LacQ140 mouse model. bioRxiv.

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Publication Preprint 2023

acylcarnitine Carbon Cardiolipins Ceramides Cholesterol Esters Coenzymes Diacylglycerol Ethers Fatty Acids Galactosylceramides Glucosylceramides Glycerophospholipids Hydroxyl Radical Lipids Lysophosphatidylcholines lysophosphatidylethanolamine lysophosphatidylinositol lysophosphatidylserine Microtubule-Associated Proteins monogalactosyldiacylglycerol Phosphates Phosphatidic Acid Phosphatidylcholines phosphatidylethanol Phosphatidylethanolamines Phosphatidylinositols Phosphatidylserines phosphoethanolamine phytosphingosine Plasmalogens prenol Sphingolipids Sphingomyelins Sphingosine Sterols Sulfoglycosphingolipids sulphoquinovosyl-diacylglycerol Triglycerides vinyl ether zymosterol

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What are the different types or variations of Glucosylceramides?

Glucosylceramides are a diverse class of glycosphingolipids that can vary in their fatty acid chain length, degree of unsaturation, and sugar moiety. Common variants include C16, C18, and C24 Glucosylceramides, as well as those with differing glycosidic linkages or substituents. Understanding these structural nuances can be important for specific applications, such as in the study of lipid rafts or signaling pathways.

How can Glucosylceramides be used in medical research and treatment?

Glucosylceramides have important implications for understanding and treating various disorders, most notably Gaucher disease. This rare lysosomal storage disorder is caused by a deficiency in the enzyme glucocerebrosidase, leading to the accumulation of Glucosylceramides. Research into Glucosylceramides can provide insights into the pathophysiology of Gaucher disease and guide the development of targeted therapies. Additionally, Glucosylceramides play a role in cellular signaling and membrane structure, making them a subject of interest for a wider range of medical applications.

What are some common challenges or considerations when working with Glucosylceramides in the lab?

Working with Glucosylceramides can present unique challenges, such as their relative insolubility in aqueous solutions and the need for specialized extraction and purification methods. Researchers must also be mindful of potential contaminants or degradation during sample handling and storage. Careful experimental design and the use of appropriate analytical techniques, such as mass spectrometry or HPLC, are crucial for accurately quantifying and characterizing Glucosylceramides in biological samples.

How can PubCompare.ai assist in Glucosylceramides research?

PubCompare.ai's AI-powered comparative platform can be a valuable tool for Glucosylceramides researchers. The platform allows you to efficiently screen protocol literature and leverage AI to pinpiont critical insights. PubCompare.ai can help you identify the most effective protocols related to Glucosylceramides, enabling you to choose the best option for reproducibility and accuracy in your studies. The platform's data-driven analysis can highlight key differences in protocol effectiveness, guiding you towards the most optimal approach for your specific research goals and unlocking the full potential of your Glucosylceramides studies.

What are some common applications of Glucosylceramides?

Glucosylceramides have a wide range of applications in biomedical research and development. They are widely used as markers for cellular differentiation and lipid raft formation. Glucosylceramides also play a role in signal transduction, cell-cell interactions, and immune function. Additionally, they have been investigated as potential therapeutic targets for various disorders, including cancer, neurodegenerative diseases, and inflammatory conditions. The versatility of Glucosylceramides makes them a subject of intense study across multiple fields of life science research.

More about "Glucosylceramides"

Glucosylceramides, also known as GlcCer, are a class of glycosphingolipids found in the plasma membrane of many cell types.
These lipid molecules play a crucial role in cellular signaling, membrane structure, and lipid raft formation.
Research into Glucosylceramides has implications for understanding and treating various disorders, including Gaucher disease, a rare lysosomal storage disorder.
To study Glucosylceramides, researchers often utilize advanced analytical techniques such as high-performance liquid chromatography (HPLC) and mass spectrometry.
The Atlantis HILIC silica column is a commonly used stationary phase for the separation of Glucosylceramides and other glycosphingolipids.
The Prominence UFLC system and AB SCIEX API 5000 mass spectrometer are examples of instrumentation that can be employed to analyze Glucosylceramide levels and profiles.
In addition to Glucosylceramides, related lipid species like Sphingomyelin are also of interest in these studies.
Analytical workflows often involve extraction and purification steps, using solvents such as Methanol and Chloroform, as well as Formic acid for appropriate sample preparation.
The PubCompare.ai platform offers an AI-powered comparative tool to help researchers easily locate the best protocols from literature, preprints, and patents, optimizing their Glucosylceramides research workflow with data-driven insights and intuitive tools.
Unlock the full potential of your Glucosylceramides studies today and uncover new avenues for understanding and treating related disorders.