Name: fatty acid 13c isotopologue profiling provides insight into trophic carbon transfer and lipid metabolism of invertebrate consumers
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Description: Watch this Scientific Journal Video about Fatty Acid 13C Isotopologue Profiling Provides Insight into Trophic Carbon Transfer and Lipid Metabolism of Invertebrate Consumers at JoVE.com

Den finansielle støtte fra R. Menzel og L. Ruess af Deutsche Forschungsgemeinschaft (RU RU780/11-1) anerkendes taknemmeligt. R. Nehring blev finansieret af RU 780/10-1. Endelig er vi yderst taknemmelige for Dr. Hazel Ruvimbo Maboreke for korrekturlæsning vores manuskript.

Beskrevet protokollen falder ikke under dyret etik kompetence. Dog når mennesker tilpasse beskrevet protokollerne til højere dyr, sørge for at den institutionelle dyr etiske komité godkendt protokollen om dyrs håndtering.
1. dyrkning af dyr
Bemærk: Alle forklaret eksperimenterende trin er baseret på veletablerede protokoller26,27,28. Biotests i laboratoriet har brug fo...

<ol>
	<li>Ruess, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Application+of+lipid+analysis+to+understand+trophic+interactions+in+soil.">Application of lipid analysis to understand trophic interactions in soil.</a> Ecology. 86 (8), 2075-2082 (2005).</li><li>Ruess, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lipid+composition+of+Collembola+and+their+food+resources+in+deciduous+forest+stands+-+Implications+for+feeding+strategies.">Lipid composition of Collembola and their food resources in deciduous forest stands - Implications for feeding strategies.</a> Soil Biology and Biochemistry. 39 (8), 1990-2000 (1990).</li><li>Chamberlain, P. M., Bull, I. D., Black, H. I. J., Ineson, P., Evershed, R. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fatty+acid+composition+and+change+in+Collembola+fed+differing+diets:+identification+of+trophic+biomarkers.">Fatty acid composition and change in Collembola fed differing diets: identification of trophic biomarkers.</a> Soil Biology and Biochemistry. 37 (9), 1608-1624 (2005).</li><li>Stott, A. W., Davies, E., Evershed, R. P., Tuross, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Monitoring+the+routing+of+dietary+and+biosynthesised+lipids+through+compound-specific+stable+isotope+(delta+C-13)+measurements+at+natural+abundance.">Monitoring the routing of dietary and biosynthesised lipids through compound-specific stable isotope (delta C-13) measurements at natural abundance.</a> Naturwissenschaften. 84 (2), 82-86 (1997).</li><li>Ruess, L., Haggblom, M. M., Langel, R., Scheu, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nitrogen+isotope+ratios+and+fatty+acid+composition+as+indicators+of+animal+diets+in+belowground+systems.">Nitrogen isotope ratios and fatty acid composition as indicators of animal diets in belowground systems.</a> Oecologia. 139 (3), 336-346 (2004).</li><li>Pollierer, M. M., Scheu, S., Haubert, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Taking+it+to+the+next+level:+Trophic+transfer+of+marker+fatty+acids+from+basal+resource+to+predators.">Taking it to the next level: Trophic transfer of marker fatty acids from basal resource to predators.</a> Soil Biology and Biochemistry. 42 (6), 919-925 (2010).</li><li>Ferlian, O., Scheu, S., Pollierer, M. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trophic+interactions+in+centipedes+(Chilopoda,+Myriapoda)+as+indicated+by+fatty+acid+patterns:+Variations+with+life+stage,+forest+age+and+season.">Trophic interactions in centipedes (Chilopoda, Myriapoda) as indicated by fatty acid patterns: Variations with life stage, forest age and season.</a> Soil Biology and Biochemistry. 52, 33-42 (2012).</li><li>Ruess, L., Chamberlain, P. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+fat+that+matters:+Soil+food+web+analysis+using+fatty+acids+and+their+carbon+stable+isotope+signature.">The fat that matters: Soil food web analysis using fatty acids and their carbon stable isotope signature.</a> Soil Biology and Biochemistry. 42 (11), 1898-1910 (2010).</li><li>Traugott, M., Kamenova, S., Ruess, L., Seeber, J., Plantegenest, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Empirically+characterising+trophic+networks:+What+emerging+DNA-based+methods,+stable+isotope+and+fatty+acid+analyses+can+offer.">Empirically characterising trophic networks: What emerging DNA-based methods, stable isotope and fatty acid analyses can offer.</a> Adv Ecol Res. 49, 177-224 (2013).</li><li>Hammer, B. T., Fogel, M. L., Hoering, T. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stable+carbon+isotope+ratios+of+fatty+acids+in+seagrass+and+redhead+ducks.">Stable carbon isotope ratios of fatty acids in seagrass and redhead ducks.</a> Chemical Geology. 152 (1-2), 29-41 (1998).</li><li>Budge, S. M., Iverson, S. J., Koopman, H. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Studying+trophic+ecology+in+marine+ecosystems+using+fatty+acids:+A+primer+on+analysis+and+interpretation.">Studying trophic ecology in marine ecosystems using fatty acids: A primer on analysis and interpretation.</a> Marine Mammal Science. 22 (4), 759-801 (2006).</li><li>Haubert, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trophic+structure+and+major+trophic+links+in+conventional+versus+organic+farming+systems+as+indicated+by+carbon+stable+isotope+ratios+of+fatty+acids.">Trophic structure and major trophic links in conventional versus organic farming systems as indicated by carbon stable isotope ratios of fatty acids.</a> Oikos. 118 (10), 1579-1589 (2009).</li><li>Ngosong, C., Raupp, J., Richnow, H. H., Ruess, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tracking+Collembola+feeding+strategies+by+the+natural+13C+signal+of+fatty+acids+in+an+arable+soil+with+different+fertilizer+regimes.">Tracking Collembola feeding strategies by the natural 13C signal of fatty acids in an arable soil with different fertilizer regimes.</a> Pedobiologia. 54 (4), 225-233 (2011).</li><li>Bec, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessing+the+reliability+of+fatty+acid-specific+stable+isotope+analysis+for+trophic+studies.">Assessing the reliability of fatty acid-specific stable isotope analysis for trophic studies.</a> Methods in Ecology and Evolution. 2 (6), 651-659 (2011).</li><li>Gladyshev, M. I., Makhutova, O. N., Kravchuk, E. S., Anishchenko, O. V., Sushchik, N. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stable+isotope+fractionation+of+fatty+acids+of+Daphnia+fed+laboratory+cultures+of+microalgae.">Stable isotope fractionation of fatty acids of Daphnia fed laboratory cultures of microalgae.</a> Limnologica. 56 (Supplement C. 56 (Supplement C), 23-29 (2016).</li><li>Gladyshev, M. I., Sushchik, N. N., Kalachova, G. S., Makhutova, O. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stable+isotope+composition+of+fatty+acids+in+organisms+of+different+trophic+levels+in+the+Yenisei+river.">Stable isotope composition of fatty acids in organisms of different trophic levels in the Yenisei river.</a> PLoS One. 7 (3), e34059 (2012).</li><li>Eisenreich, W., Dandekar, T., Heesemann, J., Goebel, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Carbon+metabolism+of+intracellular+bacterial+pathogens+and+possible+links+to+virulence.">Carbon metabolism of intracellular bacterial pathogens and possible links to virulence.</a> Nature Reviews Microbiology. 8 (6), 401-412 (2010).</li><li>Eylert, E., Bacher, A., Eisenreich, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=NMR-based+isotopologue+profiling+of+microbial+carotenoids.">NMR-based isotopologue profiling of microbial carotenoids.</a> Methods Mol Biol. 892, 315-333 (2012).</li><li>Garton, N. J., O'Hare, H. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tuberculosis:+feeding+the+enemy.">Tuberculosis: feeding the enemy.</a> Chemical Biology. 20 (8), 971-972 (2013).</li><li>Rosenblatt, J., Chinkes, D., Wolfe, M., Wolfe, R. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stable+isotope+tracer+analysis+by+GC-MS,+including+quantification+of+isotopomer+effects.">Stable isotope tracer analysis by GC-MS, including quantification of isotopomer effects.</a> Am J Physiol. 263 (3), E584-E596 (1992).</li><li>Fernandez, C. A., Des Rosiers, C., Previs, S. F., David, F., Brunengraber, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Correction+of+13C+mass+isotopomer+distributions+for+natural+stable+isotope+abundance.">Correction of 13C mass isotopomer distributions for natural stable isotope abundance.</a> J Mass Spectrom. 31 (3), 255-262 (1996).</li><li>Heuner, K., Eisenreich, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+intracellular+metabolism+of+legionella+by+isotopologue+profiling.">The intracellular metabolism of legionella by isotopologue profiling.</a> Methods Mol Biol. 954, 163-181 (2013).</li><li>Willenborg, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+the+pivotal+carbon+metabolism+of+Streptococcus+suis+serotype+2+under+ex+vivo+and+chemically+defined+in+vitro+conditions+by+isotopologue+profiling.">Characterization of the pivotal carbon metabolism of Streptococcus suis serotype 2 under ex vivo and chemically defined in vitro conditions by isotopologue profiling.</a> J Biol Chem. 290 (9), 5840-5854 (2015).</li><li>Menzel, R., Ngosong, C., Ruess, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isotopologue+profiling+enables+insights+into+dietary+routing+and+metabolism+of+trophic+biomarker+fatty+acids.">Isotopologue profiling enables insights into dietary routing and metabolism of trophic biomarker fatty acids.</a> Chemoecology. 27 (3), 101-114 (2017).</li><li>Buse, T., Ruess, L., Filser, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+trophic+biomarkers+for+Collembola+reared+on+algal+diets.">New trophic biomarkers for Collembola reared on algal diets.</a> Pedobiologia. 56 (3), 153-159 (2013).</li><li>Hutson, B. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+variations+of+the+plaster-charcoal+culture+method+on+a+Collembolan,+Folsomia+candida.">Effects of variations of the plaster-charcoal culture method on a Collembolan, Folsomia candida.</a> Pedobiologia. 18, 138-144 (1978).</li><li>Fountain, M. T., Hopkin, S. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Folsomia+candida+(Collembola):+a+&amp;#34;standard&amp;#34;+soil+arthropod.">Folsomia candida (Collembola): a &amp;#34;standard&amp;#34; soil arthropod.</a> Annu Rev Entomol. 50, 201-222 (2005).</li><li>ISO, I. O. f. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Soil Quality-Inhibition of reproduction of Collembola (Folsomia candida) by soil pollutants. , (1999).</li><li>Welch, D. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Applications+of+cellular+fatty+acid+analysis.">Applications of cellular fatty acid analysis.</a> Clin Microbiol Rev. 4 (4), 422-438 (1991).</li><li>Dodds, E. D., McCoy, M. R., Rea, L. D., Kennish, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gas+chromatographic+quantification+of+fatty+acid+methyl+esters:+flame+ionization+detection+vs.+electron+impact+mass+spectrometry.">Gas chromatographic quantification of fatty acid methyl esters: flame ionization detection vs. electron impact mass spectrometry.</a> Lipids. 40 (4), 419-428 (2005).</li><li>Kuppardt, S., Chatzinotas, A., Kastner, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+a+fatty+acid+and+RNA+stable+isotope+probing-based+method+for+tracking+protist+grazing+on+bacteria+in+wastewater.">Development of a fatty acid and RNA stable isotope probing-based method for tracking protist grazing on bacteria in wastewater.</a> Appl Environ Microbiol. 76 (24), 8222-8230 (2010).</li><li>Zhang, X., He, H., Amelung, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+GC/MS+method+for+the+assessment+of+15N+and+13C+incorporation+into+soil+amino+acid+enantiomers.">A GC/MS method for the assessment of 15N and 13C incorporation into soil amino acid enantiomers.</a> Soil Biology and Biochemistry. 39 (11), 2785-2796 (2007).</li><li>Vetter, W., Thurnhofer, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+fatty+acids+by+mass+spectrometry+in+the+selected+ion+monitoring+mode.">Analysis of fatty acids by mass spectrometry in the selected ion monitoring mode.</a> Lipid Technol. 19 (8), 184-186 (2007).</li><li>Thurnhofer, S., Vetter, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+gas+chromatography/electron+ionization-mass+spectrometry-selected+ion+monitoring+method+for+determining+the+fatty+acid+pattern+in+food+after+formation+of+fatty+acid+methyl+esters.">A gas chromatography/electron ionization-mass spectrometry-selected ion monitoring method for determining the fatty acid pattern in food after formation of fatty acid methyl esters.</a> J Agric Food Chem. 53 (23), 8896-8903 (2005).</li><li>Haubert, D., Haggblom, M. 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Isotopologue profilering
En detaljeret analyse af de kvantitative aspekter i 13C fordeling i FAs behov for avanceret teknologi til at tildele CO2 partitionering i mad webs. Den nuværende arbejde ansat isotopologue profilering for at vurdere 13C /12C nøgletal i fælles FA biomarkører for tropic interaktioner. Denne metode er veletableret aminosyre analyse af væskekromatografi (LC-MS) og blev anvendt til undersøgelser af kulstof metabolisme ...

I en kryptisk vækststeder som jord, trofiske relationer er vanskelige at adresse og er yderligere begrænset af den lille størrelse af faunaen. Det sidste tiår er sket fremskridt i biokemiske økologi, navnlig brugen af fedtsyrer som biomarkører for at definere fodringsstrategier af jord fauna under feltet betingelser1,2,3. Det skyldes at fedtsyrer fra ressourcer kan blive indarbejdet i forbrugernes væv som hele molekyler, en proces, der kaldes kosten routing4. Overførsel af fedtsyrer har været rapporteret over tre trofiske niveauer, dvs, fra svampe til nematoder til Collembola5. For nylig, den røveriske fauna blev anset for6,7 og de første anmeldelser på fedtsyrer som trofiske markører i jord mad webs har været udgivet8,9.
Mere detaljerede oplysninger om trofiske interaktioner er nået af fedtsyre stabil isotop sondering (FA-SIP). Bestemmelse af 13C /12C nøgletal i fedtsyrer i foderet og forbrugerne kan tilskrive binære links og vurdere de tilhørende kulstof flow, og har været ansat i jordbaserede, ferskvand og marine fødevarer webs10,11 ,12,13. Den grundlæggende antagelse er, at kosten dirigeres fedtsyrer ikke er omfattet af enzymatiske processer; Derfor, deres 13C signal, dvs., 13C /12C forholdet mellem fedtsyre i forbrugeren er lig den i kost1. Men en gradvis nedbrydning af 13C signatur op i fødekæden er blevet rapporteret i akvatiske systemer, derved hindrer bred anvendelse af FA-SIP i trofiske studier14,15,16. Desuden er viden i lipid metabolisme i de fleste hvirvelløse dyr i terrestriske fødevarer webs stadig begrænset.
En forståelse af lipid metabolisme veje i forbrugerne er afgørende for brugen af trofiske markør fedtsyrer som middel til bestemmelse af kvantitative carbon flow i fødekæde økologi. Med dette i tankerne, 13C-isotopologue profilering, som i princippet kan anvendes til undersøgelse af kulstof metabolismen af ethvert biologiske system17, er en lovende metode. Efter indførelsen af 13C-mærket carbon substrat, fordelingen af 13C i den metaboliske netværk er sporbar siden de genererede metaboliske produkter i forbruger viser en specifik isotopologue distribution. Dette kan vurderes ved kvantitative nukleare metaboliske resonans-spektroskopi18,19 eller massespektrometri20,21, med de sidstnævnte begunstiget i biologiske prøver med lav biomasse på grund af dets højere følsomhed.
Selvom isotopologue profilering er blevet med held anvendt til aminosyrer og givet indblik i vivo carbon metabolismen af bakterielle patogener17,22,23, dens gennemførelse i fedtsyrer syrer har haltet bagefter. Den første detaljerede analyse på skæbnen, en stabil isotop mærket forløber fedtsyre, dens kosten routing eller nedbrydning via β-oxidation, i jorden, hvirvelløse forbrugerne, blev for nylig udført af Menzel et al. 24. her, de metodologiske basics for iblanding eksperimenter med 13C mærket fedtsyrer efterfulgt af isotopologue analyse af centrale efterkommere i hyppige jord vanddyr, Collembola, leveres. Disse microarthropods er en god modelgruppe, da de udgør vigtige elementer af jord fødekæde og er godt undersøgt for deres trofiske markør fedtsyrer8,25.
En forståelse af lipid metabolisme veje i forbrugerne er afgørende for brugen af trofiske markør fedtsyrer som middel til bestemmelse af kvantitative carbon flow i fødekæde økologi. Denne protokol giver design og sæt op for et laboratorium fodring eksperiment, og de biokemiske procedurer for udtræk og methylering af dominerende lipider fraktioner (neutral lipider, fosfolipider) fra Collembola. Det viser, hvordan isotopologue sammensætning af fedtsyrer er analyseret ved massespektrometri og beskriver de relaterede formel og beregninger. Denne procedure medfører: (i) nøgletal for isotopologues overstiger massen af den overordnede ion (dvs, den fedtsyre molekylarionen M+) af en eller flere masse enheder (M+ 1M+ 2M+ 3, osv.) og (ii) samlet 13 C berigelse i fedtsyrer afledt af 13C mærket forløber. Selvom bruges til Collembola, kan denne tilgang generelt anvendes til nogen andre rovdyr-byttedyr interaktion på den forudsætning, at disse er bestemmelse i tilstrækkelige mængder under kontrollerede forhold til at sikre en vellykket etiket udbredelse og efterfølgende verifikation.

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			<td>neoLab-Round jars</td>
			<td>neoLab</td>
			<td>2-1506</td>
			<td>69 x 40 mm, 10 pacs/pack</td>
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		<tr>
			<td>Charcoal activated</td>
			<td>Carl Roth</td>
			<td>X865.1</td>
			<td>p.a., powder, CAS No. 7440-44-0</td>
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		<tr>
			<td>Alabaster Dental</td>
			<td>R&#214;HRICH-GIPSE</td>
			<td>---</td>
			<td>http://www.roehrich-gipse.de/dentalgipse.php</td>
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		<tr>
			<td>Chloroform</td>
			<td>Carl Roth</td>
			<td>7331.1</td>
			<td>HPLC &#8805; 99,9 %</td>
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		<tr>
			<td>Methanol</td>
			<td>Carl Roth</td>
			<td>P717.1</td>
			<td>HPLC &#8805; 99,9 %</td>
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		<tr>
			<td>Hexan</td>
			<td>Carl Roth</td>
			<td>7339.1</td>
			<td>HPLC &#8805; 98 %</td>
		</tr>
		<tr>
			<td>tert-Butyl methyl ether (MTBE)</td>
			<td>Carl Roth</td>
			<td>T175.1</td>
			<td>HPLC &#8805; 99,5 %</td>
		</tr>
		<tr>
			<td>Aceton</td>
			<td>Carl Roth</td>
			<td>7328.2</td>
			<td>HPLC &#8805; 99,9 %</td>
		</tr>
		<tr>
			<td>NaOH</td>
			<td>Carl Roth</td>
			<td>6771.1</td>
			<td>p.a. &#8805;99 %, in pellets</td>
		</tr>
		<tr>
			<td>di-Natriumhydrogenphosphat</td>
			<td>Carl Roth</td>
			<td>P030.1</td>
			<td>p.a. &#8805;99 % , water free</td>
		</tr>
		<tr>
			<td>Na-dihydrogenphosphat Dihydrat</td>
			<td>Carl Roth</td>
			<td>T879.1</td>
			<td>p.a. &#8805;99 %</td>
		</tr>
		<tr>
			<td>Hypochloric acid (6 N)</td>
			<td>VWR International</td>
			<td>26,115,000</td>
			<td>AVS TITRINORM vol. solution</td>
		</tr>
		<tr>
			<td>Bond Elut (Columns)</td>
			<td>Agilent Tech.</td>
			<td>14102037</td>
			<td>HF Bond Elut-SI, 500 mg, 3 mL, 50/PK</td>
		</tr>
		<tr>
			<td>Pr&#228;paratengl&#228;ser Duran</td>
			<td>Glasger&#228;tebau Ochs</td>
			<td>135215</td>
			<td>&#216; 16 x 100 mm, plus screw cap with handy knurl and integrated PTFE/silicone gasket</td>
		</tr>
		<tr>
			<td>Supelco 37 Component FAME Mix</td>
			<td>Sigma-Aldrich</td>
			<td>47885-U Supelco</td>
			<td>10&#160;mg/mL in methylene chloride, analytical standard</td>
		</tr>
		<tr>
			<td>FlowMesh</td>
			<td>Carl Roth</td>
			<td>2796.1</td>
			<td>Polypropylene mesh, approximately 0.3 mm thick, with 1 mm strand spacing</td>
		</tr>
		<tr>
			<td>Bacterial Acid Methyl Ester (BAME) Mix</td>
			<td>Sigma-Aldrich</td>
			<td>47080-U Supelco</td>
			<td>10&#160;mg/mL in methyl caproate, analytical standard</td>
		</tr>
		<tr>
			<td>Methyl nonadecanoate</td>
			<td>Sigma-Aldrich</td>
			<td>74208</td>
			<td>analytical standard &#8805; 98.0 %</td>
		</tr>
		<tr>
			<td>Hexadecanoic acid-1-13C (Palmitic)</td>
			<td>Larodan Fine Chemicals</td>
			<td>78-1600</td>
			<td>GC &#8805; 98.0 % (13C: 99.0 %)</td>
		</tr>
		<tr>
			<td>RVC 2-25 CDplus</td>
			<td>Martin Christ Gefrier-trocknungsanlagen</td>
			<td></td>
			<td>Compact benchtop midi concentrator</td>
		</tr>
		<tr>
			<td>Alpha 2-4 LDplus</td>
			<td>Martin Christ Gefrier-trocknungsanlagen</td>
			<td></td>
			<td>Drying manifold</td>
		</tr>
		<tr>
			<td>MZ 2C NT</td>
			<td>Vacuubrand GMBH</td>
			<td></td>
			<td>Vacuum pump</td>
		</tr>
		<tr>
			<td>Roto-Shake Genie</td>
			<td>Scientific Industries</td>
			<td></td>
			<td>Combined rocking and rotating device</td>
		</tr>
		<tr>
			<td>XP64 Micro Comparator</td>
			<td>Mettler Toledo</td>
			<td></td>
			<td>Super high precision balance</td>
		</tr>
		<tr>
			<td>GC-System 7890A</td>
			<td>Agilent Tech.</td>
			<td></td>
			<td>Gas chromatograph</td>
		</tr>
		<tr>
			<td>7000 GC/MS Triple Quad</td>
			<td>Agilent Tech.</td>
			<td></td>
			<td>Triple Quad mass spectrometer</td>
		</tr>
		<tr>
			<td>7683B Series Injector</td>
			<td>Agilent Tech.</td>
			<td></td>
			<td>Sample injector</td>
		</tr>
		<tr>
			<td>Heraeus Multifuge 3SR+</td>
			<td>Thermo Scientific</td>
			<td></td>
			<td>Centrifuge with 10 ml tube rotor</td>
		</tr>
	</tbody>
</table>

fatty acid 13c isotopologue profiling provides insight into trophic carbon transfer and lipid metabolism of invertebrate consumers

Fedtsyrer (FAs) er nyttige biomarkører i fødekæde økologi, fordi de er typisk ligestillet som en komplet molekyle og overføres til forbrugeren væv med mindre eller ingen ændring, så kosten routing mellem forskellige trofiske niveauer. FA trofiske markør tilgang hæmmes stadig af begrænset viden i lipid metabolisme af jord fauna. Denne undersøgelse anvendes helt mærket palmitinsyre (13C16:0, 99 atom %) som et sporstof i fedtsyre stofskifte pathways to udbredte jord Collembola, Protaphorura fimata og Heteromurus nitidus. For at undersøge den skæbne og metaboliske ændringer for dette forløber, præsenteres en metode af isotopologue profilering, udført af massespektrometri ved hjælp af enkelt ioner. Desuden, den upstream laboratorium fodring eksperiment er beskrevet, samt udvinding og methylering af dominerende lipid fraktioner (neutral lipider, fosfolipider) og den tilhørende formel og beregninger. Isotopologue profilering ikke kun udbytte den samlede 13C berigelse i fedtsyrer afledt af 13C mærket forløber men også producerer mønster af isotopologues overstiger massen af den overordnede ion (dvs. den FA molekylarionen M+) af hver mærket FA af én eller flere masse enheder (M+ 1M+ 2M+ 3, osv.). Denne viden gør det muligt for konklusioner om forholdet mellem kosten routing af en helt forbrugt FA i forhold til de novo biosyntesen. Isotopologue profilering er foreslået som et nyttigt redskab til evaluering af fedtsyre metabolisme i jord dyr at udrede trofiske interaktioner.

Frisk vægt og lipid indhold af Collembola I løbet af de beskrevne eksperiment ændre indhold i NLFAs og PLFAs ikke betydeligt over tid, den friske vægt af enheder steg lidt men ikke væsentligt24. Begge parametre angiver et godt niveau af fysisk egnethed af Collembola enheder. Vær opmærksom på at undersøge Collembolas friske vægt og lipid indhold i hele eksperimentet svarende til prøveudtagning dage for fedtsyre og isotop analyser. Bem?...

Watch this Scientific Journal Video about Fatty Acid 13C Isotopologue Profiling Provides Insight into Trophic Carbon Transfer and Lipid Metabolism of Invertebrate Consumers at JoVE.com

Fatty Acid 13C Isotopologue Profiling Provides Insight into Trophic Carbon Transfer and Lipid Metabolism of Invertebrate Consumers

Fedtsyre 13C Isotopologue profilering giver indblik i trofiske Carbon overførsel og Lipid metabolisme af hvirvelløse forbrugere

Fedtsyre trofiske markør tilgang, dvs., assimilation af fedtsyrer som hele molekylet og overførsel til forbrugeren væv med ingen eller mindre ændring, hæmmes af viden huller i fedtsyre metabolisme af små jord hvirvelløse dyr. Isotopologue profilering er fastsat som et værdifuldt redskab til at udrede trofiske interaktioner.

Fatty acids (FAs) are useful biomarkers in food web ecology because they are typically assimilated as a complete molecule and transferred into consumer ...

Research

JoVE Journal

Biochemistry

Fatty Acid 13C Isotopologue Profiling Provides Insight into Trophic Carbon Transfer and Lipid Metabolism of Invertebrate Consumers

Identification of Fatty Acids in Bacillus cereus

Identification of Fatty Acids in Bacillus cereus

Identifikation af fedtsyrer i<em&gt; Bacillus cereus</em

The Bacillus species contain branched chain and unsaturated fatty acids (FAs) with diverse positions of the methyl branch (iso or anteiso) and of the ...

Detergent-free Ultrafast Reconstitution of Membrane Proteins into Lipid Bilayers Using Fusogenic Complementary-charged Proteoliposomes.

Vaskemiddel-fri ultrahurtig rekonstituering af membranproteiner i Lipid dobbeltlag ved hjælp af Fusogenic komplementære-ladet Proteoliposomes.

Detergents are indispensable for delivery of membrane proteins into 30-100 nm small unilamellar vesicles, while more complex, larger model lipid bilayers ...

Sampling and Pretreatment of Tooth Enamel Carbonate for Stable Carbon and Oxygen Isotope Analysis

Prøveudtagning og forbehandling af tand emalje carbonat for stabil Carbon og Oxygen isotop analyser

Stable carbon and oxygen isotope analysis of human and animal tooth enamel carbonate has been applied in paleodietary, paleoecological, and ...

Nitropeptide Profiling and Identification Illustrated by Angiotensin II

Nitropeptid profilering og identifikation illustreret af angiotensin II

Protein nitration is one of the most important post-translational modifications (PTM) on tyrosine residues and it can be induced by chemical actions of ...

Isolation of Lipoprotein Particles from Chicken Egg Yolk for the Study of Bacterial Pathogen Fatty Acid Incorporation into Membrane Phospholipids

Isolering af lipoprotein partikler fra hønseæg æggeblomme til undersøgelse af bakteriel Patogenchlorid fedtsyre inkorporering i membran Fosfolipids

Staphylococcus aureus and other Gram-positive pathogens incorporate fatty acids from the environment into membrane phospholipids. During infection, the ...

Detergent-assisted Reconstitution of Recombinant Drosophila Atlastin into Liposomes for Lipid-mixing Assays

Detergent-assisted Reconstitution of Recombinant Drosophila Atlastin into Liposomes for Lipid-mixing Assays

Vaskemiddel-assisteret rekonstitution af rekombinant Drosophila Atlastin til Liposomer til lipid-blanding assays

Membrane fusion is a crucial process in the eukaryotic cell. Specialized proteins are necessary to catalyze fusion. Atlastins are endoplasmic reticulum ...

Profiling Ubiquitin and Ubiquitin-like Dependent Post-translational Modifications and Identification of Significant Alterations

Profilering af Ubiquitin og Ubiquitin-lignende afhængige posttranslationelle ændringer og identificering af væsentlige ændringer

Ubiquitin (ub) and ubiquitin-like (ubl) dependent post-translational modifications of proteins play fundamental biological regulatory roles within the ...

Optimized Incorporation of Alkynyl Fatty Acid Analogs for the Detection of Fatty Acylated Proteins using Click Chemistry

Optimeret inkorporering af alkynylfedtsyreanaloger til påvisning af fedtaylerede proteiner ved hjælp af klikkemi

Fatty acylation, the covalent addition of saturated fatty acids to protein substrates, is important in regulating a myriad of cellular functions in ...

Ganglioside Extraction, Purification and Profiling

Gangliosidekstraktion, rensning og profilering

Gangliosides are glycosphingolipids that contain one or more sialic acid residues. They are found on all vertebrate cells and tissues but are especially ...