Name: a method for extracting pigments from squid doryteuthis pealeii
Uploaded: 2016-11-09T15:00:00
Description: Watch this Scientific Journal Video about A Method for Extracting Pigments from Squid Doryteuthis pealeii at JoVE.com

The authors gratefully acknowledge the use of facilities at the University of New Hampshire including the University Instrumentation Center. This work was supported by the University of New Hampshire, Department of Chemistry.

Hvirvelløse dyr undersøgelser heri er ikke reguleret i USA; derfor Institutional Animal Care og brug Udvalg har ingen myndighed til nyvurdering af sådanne protokoller. I stedet for disse ikke falder ind under den kompetence, regulering i USA, forfatterne erklærer, at disse undersøgelser blev udført med oprigtig indsats mod etiske brug, pleje og behandling af disse dyr, antallet af individer blev minimeret, og disse bestræbelser er i overensstemmelse med Basel-erklæringen og det Internationale Råd for Laboratory Animal Science (ICLA...

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Vi har vist en metode til at udvinde pigmenter fra blæksprutte chromatophore granulat. Ved specifikt at målrette granulatet, vores mål er at bestemme deres rolle i at mediere adaptiv farve. Denne metode adskiller sig fra tidligere rapporter konstrueret til at specificere blæksprutter pigmenter hjælp bulk-vævsprøver 14 eller frysetørret hud 15. Mens denne protokol er effektiv til at udvinde chromatophore pigmenter, er det begrænset til små reaktion skalaer. Dis...

Blæksprutter såsom blæksprutte, blæksprutter og blæksprutte har evnen til dynamisk at ændre deres udseende til at camouflere og signalering. 1-6 Denne kapacitet er delvist understøttet af den selektive areal udvidelse af pigmenterede organer kendt som chromatophores. 4,7-9 chromatophores er bløde aktuatorer, der indeholder et netværk af nanostrukturerede pigment granulat begrænset inden for en cytoelastic sacculus der er forankret radialt ved muskelfibre. 1,3 Da de er aktiveret, chromatophores vokse med 500% i præsenteret overfladeareal distribuere granulatet i hele orglet. 3,7 , 10,11 Når denne handling er samordnet over et antal chromatophores, er den samlede farvning af dyret ændret. Mens det er kendt, at pigmentgranula bidrager til denne farveændring, deres sammensætning forbliver ukendt. Vi beskriver en fremgangsmåde til at isolere og oprense chromatophore pigmenter, der kan tilpasses til fremtidige kompositionelle studier. 3,12 Chromatophore indeholder væv høstes gennem omhyggelig udryddelse fra blæksprutter. En fordøjelse og homogenisering proces anvendes derefter til at dissociere det omgivende væv og adskille chromatophore celler. De nanostrukturerede granuler isoleres derefter og oprenses fra de resterende chromatophores anvendelse af gentagne sonikering og centrifugering. Efter oprensning, er pigmenter udvundet fra granulerne i en proces, der er tilpasset fra udvinding af synlig farve fra sommerfuglevinger anvendelse af sure methanolopløsninger. 13 Scanning elektronmikroskopi (SEM) og spektrofotometri anvendes til at bekræfte, at de chromatophore pigmenter held ekstraheres under anvendelse denne proces. Denne metode beskriver isolering af chromatophore granulater som bruges til at udforske de molekylære bidrag til coloration i blæksprutter. 12 Små molekyler ekstraktioner fra hele dyr kan ofte være en lang og kedelig proces. Målet her er at informere kommende forskere i en effektiv og letkøbt protokol om erhvervelse af pigmenter fra nanostrukturerede granulat i blæksprutter.

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			<td height="42" style="height:42px;width:216px;">Collagenase</td>
			<td style="width:71px;">Alfa Aesar</td>
			<td style="width:119px;">9001-12-1</td>
			<td style="width:205px;">No hazard</td>
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			<td height="42" style="height:42px;width:216px;">Dithiothreitol</td>
			<td style="width:71px;">Sigma-Aldrich</td>
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			<td>Irritant, acute toxicity</td>
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			<td height="42" style="height:42px;width:216px;">HEPES</td>
			<td style="width:71px;">Sigma-Aldrich</td>
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			<td>Mild irritant</td>
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			<td height="63" style="height:63px;width:216px;">Hydrochloric acid</td>
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			<td>Reacts violently with oxidants</td>
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			<td>Mild eye irritant</td>
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			<td height="42" style="height:42px;width:216px;">Methanol</td>
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			<td height="42" style="height:42px;width:216px;">Papain</td>
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a method for extracting pigments from squid doryteuthis pealeii

Cephalopods can undergo rapid and adaptive changes in dermal coloration for sensing, communication, defense, and reproduction purposes. These capabilities are supported in part by the areal expansion and retraction of pigmented organs known as chromatophores. While it is known that the chromatophores contain a tethered network of pigmented granules, their structure-function properties have not been fully detailed. We describe a method for isolating the nanostructured granules in squid Doryteuthis pealeii chromatophores and demonstrate how their associated pigments can be extracted in acidic solvents. To accomplish this, the chromatophore containing dermal layer is first manually isolated using a superficial dissection, and the pigment granules are removed using sonication, centrifugation, and washing cycles. Pigments confined within the purified granules are then extracted via acidic methanol solutions, leaving nanostructures with smaller diameters that are void of visible color. This extraction procedure produces a 58% yield of soluble pigments isolated from granules. Using this method, the composition of the chromatophore pigments can be determined and used to provide insight into the mechanism of adaptive coloration in cephalopods.

Chromatophores dissekeres fra D. pealeii dorsale kappe (figur 1A, 1B). Når de først er fjernet, er chromatophores lyseret, og oprenset ved anvendelse af centrifugering og vaskecyklusser at isolere de pigmenterede granuler (figurerne 2A, 2B). Sure methanolopløsninger (HCl-MeOH) anvendes til at ekstrahere pigmentet fra granulatet (figur 2C), hvilket gav et opløseligt pigment ekstrakt og uoplø...

Watch this Scientific Journal Video about A Method for Extracting Pigments from Squid Doryteuthis pealeii at JoVE.com

A Method for Extracting Pigments from Squid Doryteuthis pealeii

En protokol til ekstraktion af pigmenter fra nanostrukturerede granulat i blæksprutte Doryteuthis pealeii chromatophores præsenteres.

En fremgangsmåde til ekstraktion Pigmenter fra Squid<em&gt; Doryteuthis pealeii</em

Cephalopods can undergo rapid and adaptive changes in dermal coloration for sensing, communication, defense, and reproduction purposes. These capabilities ...

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Biology

A Method for Extracting Pigments from Squid Doryteuthis pealeii

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