Name: measurement of mitochondrial oxygen consumption in permeabilized fibers of drosophila using minimal amounts of tissue
Uploaded: 2018-04-07T14:00:00
Description: Watch this Scientific Journal Video about Measurement of Mitochondrial Oxygen Consumption in Permeabilized Fibers of Drosophila Using Minimal Amounts of Tissue at JoVE.com

Undersøgelsen blev finansieret af tilskud fra National Sciences og Engineering Research Council (NSERC, discovery grant) og Université de Moncton til NP. LHB vil gerne anerkende den finansielle støtte fra det canadiske Institut for sundhed Research (CIHR), New Brunswick Innovation Foundation (NBIF) og Université de Moncton. Arbejdet i EHC understøttes af Alzheimers Society of Canada, hjernen Canada, NSERC, canadiske Breast Cancer Foundation, New Brunswick Innovation Foundation, New Brunswick sundhed Research Foundation og Université de Moncton.

1. reagenser forberedelse
<ol><li>Forberede følgende løsninger for dissektion og permeabilization af vævet.<ol>
<li>Forberede bevarelse løsning: 2.77 CaK2EGTA, 7,23 mM K2EGTA, 5.77 mM Na2ATP, 6.56 mM MgCl2, 20 mM taurin, 15 mM Na2fosforkreatin, 20 mM imidazol, 0,5 mM dithiothreitol, og 50 mM K-MES, pH 7.1 (kan gemmes ved-20 ° C).</li>
		<li>Forberede saponin løsning: 5 mg af saponin i 1 mL af bevarelse løsning (forberede frisk dagligt).</li>
	</ol>
</li>
	<l...

<ol>
	<li>Stephenson, R., Metcalfe, N. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drosophila+melanogaster:+a+fly+through+its+history+and+current+use.">Drosophila melanogaster: a fly through its history and current use.</a> The journal of the Royal College of Physicians of Edinburgh. 43 (1), 70-75 (2013).</li><li>Morgan, T. 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T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+of+Parkinson's+disease.">Animal models of Parkinson's disease.</a> FEBS Journal. 279 (7), 1156-1166 (2012).</li><li>Baker, K. D., Thummel, C. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diabetic+Larvae+and+Obese+Flies-Emerging+Studies+of+Metabolism+in+Drosophila.">Diabetic Larvae and Obese Flies-Emerging Studies of Metabolism in Drosophila.</a> Cell Metabolism. 6 (4), 257-266 (2007).</li><li>Morris, S. N. 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U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Naturally+occurring+mitochondrial+dna+haplotypes+exhibit+metabolic+differences:+insight+into+functional+properties+of+mitochondria.">Naturally occurring mitochondrial dna haplotypes exhibit metabolic differences: insight into functional properties of mitochondria.</a> Evolution. 66 (10), 3189-3197 (2012).</li><li>Pichaud, N., Messmer, M., Correa, C. C., Ballard, J. W. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diet+influences+the+intake+target+and+mitochondrial+functions+of+Drosophila+melanogaster+males.">Diet influences the intake target and mitochondrial functions of Drosophila melanogaster males.</a> Mitochondrion. 13 (6), 817-822 (2013).</li><li>Wolff, J. N., Pichaud, N., Camus, M. F., C&#244;t&#233;, G., Blier, P. U., Dowling, D. 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I denne undersøgelse, er en metode til prøveforberedelse forud for målingerne af mitokondrie iltforbrug i Drosophila beskrevet. Denne metode blev udviklet til at løse forskellige problemer relateret til protokoller ved hjælp af mitokondrie isolering, navnlig med hensyn til varighed og antal individer kræves. Stedet for at arbejde med mitokondrie isolering der normalt kræver store beløb af væv fra flere personer, er dette eksperiment udført på permeabilized muskelfibre fra thoraxes af par Drosophila. I denne pr...

Bananfluen Drosophila melanogaster, har været brugt som en model organisme for genetisk forskning for over et århundrede1. Studiet af denne organisme har ikke kun ført til betydelig grundlæggende viden om kønsbunden nedarvning2, mutation sats3, udvikling af neurale system og celle skæbne bestemmelse4, men har også for nylig dukket op som en værdifuldt redskab til at studere de mekanismer, der er forbundet med flere sygdomme som Alzheimers og Parkinsons5,6. Derudover er det en populær model til at studere aldringsprocessen, da de kan blive rejst i stort antal over en kort periode og har en kort levetid. De også besidder homolog strukturer til menneskelige organer, som et hjerte, oenocytes (hepatocyt-lignende celler), fedt organer (fungerer på samme måde som den lever og hvidt fedtvæv), insulin producerende celler (svarende til β-pancreas celler), samt den hemolymph transporterer metabolitter (analog med blod fra hvirveldyr)7. Desuden er de centrale veje i formidlende stofskifte (herunder insulin/insulin-lignende vækstfaktor-lignende signalvejen og målet på Rapamycin-TOR veje) også meget bevaret7. Af disse grunde Drosophila er for nylig blevet udnyttet til at beskrive de grundlæggende mekanismer, der styrer stofskiftet, især i patologiske tilstande forbundet med menneskelige metaboliske sygdomme som diabetes8. En større komponent af stofskiftet er mitokondriet, der integrerer flere veje og udfører en af livets mest vigtige biologiske funktioner, ATP produktion, via oxidativ fosforylering proces (OXPHOS). I betragtning af deres centrale rolle i organismes metabolisme er det ikke overraskende, at mitokondriel dysfunktioner er involveret i mange sygdomme som Parkinsons9 og Alzheimers sygdomme10samt Amyotrofisk lateral sklerose 11 , 12. de er også grundlæggende determinanter for aldringsprocessen. De er de største producenter af reaktive ilt arter (ROS) i den celle, som kan være skadelig for cellen i høj koncentration gennem oxidative skader11. Aldring er også blevet forbundet til ophobning af beskadigede eller muterede mitokondriel DNA13, mitophagy dysfunktioner14,15 samt værdiforringelse af mitokondriel Biogenese16. Mitokondrier er også vigtige determinanter af cellens homøostase, som de kan anvende forskellige substrater for at justere flere cellulære funktioner henhold til overflod og knaphed på makronæringsstoffer17,18.
Faktisk, de forskellige næringsstoffer i kosten (kulhydrater, lipider og proteiner) er fordøjet, absorberet og transporteret i cellerne. De er så forvandlet i i cytosol, og de afledte substrater er transporteret ind i mitokondriets matrix, hvor de producerer reducerende ækvivalenter, såsom NADH og FADH219. Disse reducerende ækvivalenter oxideres derefter af forskellige enzymatiske komplekser af elektronen transportsystem (ETS). Disse komplekser er indlejret i den indre membran, som kompleks I og komplekse II. Derudover repræsenterer andre enzymatiske komplekser såsom den mitokondrielle glycerol-3-fosfat dehydrogenase og prolin dehydrogenase alternative ruter til indtastning af elektroner i ETS20,21. Disse «alternative» komplekser er særlig vigtig i insekter, som alt efter dyreart, de kan deltage aktivt for at øge respiration20,22,23,21. Elektroner fra disse ETS fodersystemer overføres til ubikinon og efterfølgende til komplekse III, og derefter til komplekse IV, indtil den endelige acceptor, Molekylær ilt. Denne elektron overførsel genererer proton-drivkraften i hele den indre mitokondrielle membran kørsel fosforylering af ADP at ATP på komplekse V (figur 1). Overvejer de centrale rolle af mitochondrier i cellen homøostase, studerer mitokondrie stofskifte ved hjælp af den relevante model D. melanogaster repræsenterer et kraftfuldt værktøj til at afgrænse de underliggende mekanismer af forskellige patofysiologiske betingelser eller under cellulære og miljømæssige belastninger. Overraskende men målt kun en håndfuld af undersøgelser faktisk mitokondrie respiration i Drosophila24,25,26. Faktisk, eksperimenter med henblik på at evaluere mitokondrie iltforbrug kræver isolering af mitokondrier. Selv om fordelagtige for måling af forskellige mitokondriets funktioner (f.eks. ROS produktion eller P/O ratio som markør af mitokondrie effektivitet27,28), kræver disse isolering generelt temmelig store mængder væv fra flere personer24,29. Dette krav om høje mængder af væv og enkeltpersoner er en vigtig begrænsende faktor, især i betragtning af at alle individer skal være samme alder og helst af samme køn for eksperimenterne, at foranstaltningen af åndedræt på andet tidspunkt peger besværlige i bedste fald. Desuden mens mitokondrie isolering kan give betydelig indsigt i de grundlæggende mekanismer, der regulerer mitokondrie metabolisme, har de metoder, der anvendes til at isolere mitokondrier flere ulemper som er vanskeligheden at opnå reproducerbare resultater , afbrydelse af mitokondrie netværk og ændring af mitokondrie struktur og funktion29,30,31.
Formålet med denne undersøgelse er at præsentere en robust protokol for at måle mitokondrie iltforbrug i Drosophila ved hjælp af kun en minimal mængde af væv fra meget få personer. Denne protokol består af måling af mitokondrie ilt forbrug i situ ved permeabilized muskelfibre29 fra Drosophila thoraxes i kombination med høj opløsning respirometri32,33, 34 , 35. denne metode har også yderligere fordele i forhold til den klassiske mitokondrie isolation metode da interaktioner med andre komponenter i cellen som godt som mitokondrielle struktur og funktion er mere bevaret i permeabilized fibre29,31,36, hvilket gør denne metode mere fysiologisk relevante. Med denne protokol, kan mitokondrie funktioner præcist evalueres ved hjælp af høj opløsning respirometri i kun tre thoraxes i Drosophila, med substrater at tillade bestemmelse af iltforbrug på flere forskellige trin af ETS. Derfor kunne denne protokol hjælpe svar vigtige spørgsmål om de grundlæggende mekanismer, der styrer stofskiftet i forbindelse med mange miljømæssige eller patofysiologiske forhold ved at udnytte Drosophila modellen.
At måle iltforbruget på flere forskellige trin af ETS og vurdere, hvordan forskellige substrater bidrage til respiration, forskellige substrater (figur 1), uncoupler og -hæmmere er brugt30 efter permeabilization af den væv. Specifikt, er sekventiel tilføjelser af forskellige substrater udført for at stimulere optagelse af elektroner gennem forskellige komplekser af ETS. En uncoupler, carbonyl cyanid 4-(trifluormethoxy) phenylhydrazone (FCCP), derefter tilsættes på optimal koncentration for at måle den ikke-kombineret respiration, dvs. den ikke-phosphorylating respiration stimuleret til maksimal iltforbrug. Sekventiel hæmninger af komplekser I, II og III er derefter udført for at overvåge det resterende iltforbrug, der er på grund af ikke-ETS oxidation reaktioner. Endelig, komplekse IV maksimal respiration kapacitet kan evalueres ved injektion af N, N, N', Nielsen, - Tetramethyl - p-phenylendiamin (TMPD), en kunstig elektron udbyder, og Ascorbat. Det er vigtigt at bemærke, at forsøgene er udført på 24 °C, da det er den temperatur, hvor fluerne er rejst.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>High-resolution respirometer Oxygraph O2K</td>
			<td>Oroboros Instruments, Innsbruck, Austria</td>
			<td>10022-02</td>
			<td>Startup O2K respirometer kit 
			&#160;</td>
		</tr>
		<tr>
			<td>O2K-Titration Set&#160;</td>
			<td>Oroboros Instruments, Innsbruck, Austria</td>
			<td>20820-03</td>
			<td>Hamilton syringes with different volumes 
			&#160;</td>
		</tr>
		<tr>
			<td>Datlab software</td>
			<td>Oroboros Instruments, Innsbruck, Austria</td>
			<td>20700</td>
			<td>Software for data acquisition and analysis 
			&#160;</td>
		</tr>
		<tr>
			<td>Fine-tipped antimagnetic forceps</td>
			<td>VWR</td>
			<td>82027-400</td>
			<td> 
			&#160;</td>
		</tr>
		<tr>
			<td>Secura225D-1S-DQE</td>
			<td>Sartorius AG, Goettingen, Germany</td>
			<td></td>
			<td>Semi-micro balance (distributed by several companies)&#160; 
			&#160;</td>
		</tr>
		<tr>
			<td>Drosophila melanogaster wild-type w1118</td>
			<td>Bloomington Drosophila stock Center, IN, USA</td>
			<td></td>
			<td> 
			Storage Condition: 24 &#176;C</td>
		</tr>
		<tr>
			<td>Ethylene glycol-bis(2-aminoethylether)-N,N,N&#8242;,N&#8242;-tetraacetic acid</td>
			<td>Sigma-Aldrich</td>
			<td>E4378</td>
			<td>EGTA 
			Storage Condition: RT</td>
		</tr>
		<tr>
			<td>KOH</td>
			<td>Sigma-Aldrich</td>
			<td>P1767</td>
			<td>CAUTION: corrosive to metals, acute toxicity, skin corrosion, serious eye damage, acute aquatic toxicity. 
			Storage Condition: RT</td>
		</tr>
		<tr>
			<td>CaCO3</td>
			<td>Sigma-Aldrich</td>
			<td>C4830</td>
			<td> 
			Storage Condition: RT</td>
		</tr>
		<tr>
			<td>Na2ATP</td>
			<td>Sigma-Aldrich</td>
			<td>A2383</td>
			<td> 
			Storage Condition: -20 &#176;C</td>
		</tr>
		<tr>
			<td>MgCl2.6H2O</td>
			<td>Sigma-Aldrich</td>
			<td>M9272</td>
			<td> 
			Storage Condition: RT</td>
		</tr>
		<tr>
			<td>Taurine</td>
			<td>Sigma-Aldrich</td>
			<td>T0625</td>
			<td> 
			Storage Condition: RT</td>
		</tr>
		<tr>
			<td>Na2Phosphocreatine</td>
			<td>Sigma-Aldrich</td>
			<td>P7936</td>
			<td> 
			Storage Condition: -20 &#176;C</td>
		</tr>
		<tr>
			<td>Imidazole</td>
			<td>Sigma-Aldrich</td>
			<td>I5513</td>
			<td> 
			Storage Condition: RT</td>
		</tr>
		<tr>
			<td>Dithiothreitol</td>
			<td>Sigma-Aldrich</td>
			<td>D0632</td>
			<td> 
			Storage Condition: 2-8 &#176;C</td>
		</tr>
		<tr>
			<td>MES hydrate</td>
			<td>Sigma-Aldrich</td>
			<td>M8250</td>
			<td> 
			Storage Condition: RT</td>
		</tr>
		<tr>
			<td>Saponin from quillaja bark</td>
			<td>Sigma-Aldrich</td>
			<td>S7900</td>
			<td>Saponin 
			Storage Condition: RT 
			Solution Preparation: 5 mg in 1 mL of preservation solution. Prepare fresh daily.</td>
		</tr>
		<tr>
			<td>KCl</td>
			<td>Sigma-Aldrich</td>
			<td>P9541</td>
			<td> 
			Storage Condition: RT</td>
		</tr>
		<tr>
			<td>KH2PO4</td>
			<td>Sigma-Aldrich</td>
			<td>P9791</td>
			<td> 
			Storage Condition: RT</td>
		</tr>
		<tr>
			<td>HEPES</td>
			<td>Sigma-Aldrich</td>
			<td>H3375</td>
			<td> 
			Storage Condition: RT</td>
		</tr>
		<tr>
			<td>BSA</td>
			<td>Sigma-Aldrich</td>
			<td>05470</td>
			<td> 
			Storage Condition: 2-8 &#176;C</td>
		</tr>
		<tr>
			<td>Na2S2O4</td>
			<td>Sigma-Aldrich</td>
			<td>157953</td>
			<td>Sodium dithionite. CAUTION: self-heating substances and mixtures, acute toxicity, acute aquatic toxi chronic aquatic toxicity. 
			Storage Condition: RT</td>
		</tr>
		<tr>
			<td>Sodium pyruvate</td>
			<td>Sigma-Aldrich</td>
			<td>P2256</td>
			<td>Pyruvate 
			Storage Condition: 2-8 &#176;C 
			Solution Preparation: In MilliQ water. Prepare fresh daily.</td>
		</tr>
		<tr>
			<td>L-(-)-Malic acid</td>
			<td>Sigma-Aldrich</td>
			<td>M1000</td>
			<td>Malate 
			Storage Condition: RT 
			Solution Preparation: In MilliQ water. Neutralize with KOH and store at -20 &#176;C.</td>
		</tr>
		<tr>
			<td>Adenosine 5&#39;-diphosphate monopotassium salt hydrate</td>
			<td>Sigma-Aldrich</td>
			<td>A5285</td>
			<td>ADP 
			Storage Condition: -20 &#176;C 
			Solution Preparation: In MilliQ water. Neutralize with KOH and store at -80 &#176;C.</td>
		</tr>
		<tr>
			<td>Cytochrome c from equine heart</td>
			<td>Sigma-Aldrich</td>
			<td>C7752</td>
			<td>Cytochrome c 
			Storage Condition: -20 &#176;C 
			Solution Preparation: In MilliQ water. Store at -20 &#176;C.</td>
		</tr>
		<tr>
			<td>L-Proline</td>
			<td>Sigma-Aldrich</td>
			<td>P0380</td>
			<td>Proline 
			Storage Condition: RT 
			Solution Preparation: In MilliQ water. Store at -20 &#176;C.</td>
		</tr>
		<tr>
			<td>Sodium succinate dibasic hexahydrate</td>
			<td>Sigma-Aldrich</td>
			<td>S2378</td>
			<td>Succinate 
			Storage Condition: RT 
			Solution Preparation: In MilliQ water. Neutralize with HCl and store at -20 &#176;C.</td>
		</tr>
		<tr>
			<td>sn-Glycerol 3-phosphate bis(cyclohexylammonium) salt</td>
			<td>Sigma-Aldrich</td>
			<td>G7886</td>
			<td>Glycerol-3-phosphate 
			Storage Condition: -20 &#176;C 
			Solution Preparation: In MilliQ water. Neutralize with HCl and store at -80 &#176;C.</td>
		</tr>
		<tr>
			<td>Carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone</td>
			<td>Sigma-Aldrich</td>
			<td>C2920</td>
			<td>FCCP. CAUTION: acute toxicity, skin sensitisation, chronic aquatic toxicity. 
			Storage Condition: RT 
			Solution Preparation: In absolute ethanol. Store in glass vials at -20 &#176;C.</td>
		</tr>
		<tr>
			<td>Rotenone</td>
			<td>Sigma-Aldrich</td>
			<td>R8875</td>
			<td>CAUTION: acute toxicity, skin irritation, eye irritation, specific target organ toxicity (respir sytem), acute aquatic toxicity, chronic aquatic toxicity. 
			Solution Preparation: In absolute ethanol. Store in dark vials at -20 &#176;C.</td>
		</tr>
		<tr>
			<td>Malonic acid</td>
			<td>Sigma-Aldrich</td>
			<td>M1296</td>
			<td>Malonate. CAUTION: acute toxicity, serious eye damage. 
			Storage Condition: RT 
			Solution Preparation: In MilliQ water. Neutralize with KOH. Prepare fresh daily.</td>
		</tr>
		<tr>
			<td>Antimycin A from Streptomyces sp.</td>
			<td>Sigma-Aldrich</td>
			<td>A8674</td>
			<td>Antimycin A. CAUTION: acute toxicity, acute aquatic toxicity, chronic aquatic toxicity. 
			Storage Condition: -20 &#176;C 
			Solution Preparation: In absolute ethanol. Store at -20 &#176;C.</td>
		</tr>
		<tr>
			<td>N,N,N&#8242;,N&#8242;-Tetramethyl-p-phenylenediamine</td>
			<td>Sigma-Aldrich</td>
			<td>T7394</td>
			<td>TMPD 
			Storage Condition: RT 
			Solution Preparation: In MilliQ water. Store in dark vials at&#160; -20 &#176;C.</td>
		</tr>
		<tr>
			<td>(+)-Sodium L-ascorbate</td>
			<td>Sigma-Aldrich</td>
			<td>A4034</td>
			<td>Ascorbate 
			Storage Condition: RT 
			Solution Preparation: In MilliQ water. Store in dark vials at&#160; -20 &#176;C.</td>
		</tr>
		<tr>
			<td>NaN3</td>
			<td>Sigma-Aldrich</td>
			<td>S2002</td>
			<td>Sodium azide. CAUTION: acute toxicity (oral and dermal), specific target organ toxicity (brain), aquatic toxicity, chronic aquatic toxicity.&#160; 
			Solution Preparation: In MilliQ water. Store at -20 &#176;C.</td>
		</tr>
	</tbody>
</table>

measurement of mitochondrial oxygen consumption in permeabilized fibers of drosophila using minimal amounts of tissue

Frugtflue, Drosophila melanogaster, repræsenterer en spirende model for studiet af stofskiftet. Faktisk, drosophila har strukturer homologe til menneskelige organer, besidder meget bevaret stofskifteveje og har en relativt kort levetid, der giver mulighed for undersøgelse af forskellige grundlæggende mekanismer i en kort periode. Det er imidlertid overraskende, at en af de mekanismer, der er afgørende for cellulære stofskifte, den mitokondrielle respiration, ikke er blevet grundigt undersøgt i denne model. Det er sandsynligvis fordi foranstaltning af den mitokondrielle respiration i Drosophila kræver som regel et meget stort antal enkeltpersoner og de opnåede resultater er ikke meget reproducerbare. Her, er en metode, der giver den præcise måling af mitokondrie iltforbrug ved hjælp af minimale mængder af væv fra Drosophila beskrevet. I denne metode, er thoraxes dissekeret og permeabilized både mekanisk med skarpe pincet, og kemisk saponin, giver mulighed for forskellige stoffer til at krydse cellemembranen og modulere den mitokondrielle respiration. Efter permeabilization, er en protokol, der udført for at evaluere de forskellige komplekser af elektronen transportsystem (ETS) evne til at oxidere forskellige substrater, samt deres svar på en uncoupler og på flere hæmmere. Denne metode indebærer mange fordele i forhold til metoder ved hjælp af mitokondrie isolering, som det er mere fysiologisk relevante fordi mitokondrier stadig interagerer med andre cellulære komponenter og mitokondriel morfologi er bevaret. Desuden prøve præparater er hurtigere, og de opnåede resultater er yderst reproducerbare. Ved at kombinere fordelene ved Drosophila som en model for studiet af stofskifte med evalueringen af mitokondrie respiration, vigtige nye indsigter kan blive afsløret, især hvor fluerne oplever forskellige miljømæssige eller patofysiologiske betingelser.

En repræsentativ spor af mitokondrie iltforbrug ved hjælp af den protokol, der er beskrevet ovenfor er fastsat i figur 2. Pyruvat og malat injiceres i kamre sammen med de permeabilized muskelfibre er henvist til CI-lækage respiration, dvs., når komplekset af Emissionshandelsordningen er stimuleret af NADH produceret gennem oxidation af pyruvat og malat via den tricarboxylic syre cyklus (CI). Under denne åndedræt sats opretholdes den mitokondrie...

Watch this Scientific Journal Video about Measurement of Mitochondrial Oxygen Consumption in Permeabilized Fibers of Drosophila Using Minimal Amounts of Tissue at JoVE.com

Measurement of Mitochondrial Oxygen Consumption in Permeabilized Fibers of Drosophila Using Minimal Amounts of Tissue

I dette papir, er der beskrevet en metode til at måle iltforbrug ved hjælp af høj opløsning respirometri i permeabilized thoraxes i Drosophila. Denne teknik kræver en minimal mængde af væv i forhold til den klassiske mitokondrie isolation teknik og de opnåede resultater er mere fysiologisk relevante.

Måling af mitokondrie iltforbrug i Permeabilized fibre fra Drosophila bruger minimale mængder af væv

The fruit fly, Drosophila melanogaster, represents an emerging model for the study of metabolism. Indeed, drosophila have structures homologous to human ...

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Using Scaffold Liposomes to Reconstitute Lipid-proximal Protein-protein Interactions In Vitro

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