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.

Virvelløse dyr studier utført her, er ikke regulert i USA; derfor Institutional Animal Care og bruk komité har ingen myndighet for vurdering av slike protokoller. I stedet for disse ikke faller inn under jurisdiksjonen til regulering i USA, forfatterne herved oppgir at disse studiene ble utført med oppriktig innsats mot etisk bruk, omsorg og behandling av disse dyrene, antall individer ble minimert og dette arbeidet er i overensstemmelse med Basel-erklæringen og det internasjonale råd for Laboratory Animal Science (ICLAS) etiske retni...

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Vi har vist en metode for å utvinne pigmenter fra blekksprut chromatophore granulat. Av spesiell fokus på granulater, er vårt mål å bestemme deres rolle som formidler adaptive farge. Denne metoden skiller seg fra tidligere rapporter utformet for å karakterisere Cephalopod pigmenter bruker bulk vevsprøver 14 eller frysetørket hud 15. Mens denne protokollen er effektiv ved utpakking chromatophore pigmenter, er det begrenset til små reaksjons skalaer. Dissekere en...

Blekkspruter som blekksprut, blekksprut og blekksprut har evnen til å dynamisk endre sitt utseende for å kamuflere og signalering. 1-6 Denne funksjonen støttes delvis av den selektive areal utvidelse av pigment organer kjent som kromatofor. 4,7-9 kromatofor er myke aktuatorer som inneholder et nettverk av nanostrukturerte pigment granulater trange innenfor en cytoelastic sacculus som er forankret radialt av muskelfibre. 1,3 Ettersom de er aktivert, kromatofor utvide med 500% i present areal å fordele granulater hele organ. 3,7 , 10,11 Når denne handlingen er samordnet over et antall kromatofor, blir den samlede fargen av dyret endret. Mens det er kjent at pigmentgranulater bidra til denne fargeendring, forblir deres sammensetning ukjent. Vi beskriver en fremgangsmåte for å isolere og rense chromatophore pigmenter som kan tilpasses for fremtidige komposisjonelle studier. 3,12 chromatophore inneholder vev er høstet gjennom forsiktig extirpation fra cephalopod. En fordøyelse og homogenisering prosess blir så brukt for å dissosiere det omgivende vev og skille de chromatophore cellene. Den nanostrukturerte granuler blir så isolert og renset fra de gjenværende kromatofor ved hjelp av gjentatt, ultralydbehandling og sentrifugering. Etter rensing blir pigmenter ekstrahert fra granuler i en prosess som er tilpasset etter utvinning av synlig farge fra kulisser ved hjelp av sure metanolløsninger. 13 Scanning elektronmikroskopi (SEM) og spektrofotometri blir anvendt for å bekrefte at chromatophore pigmentene med hell ekstraheres ved hjelp denne prosessen. Denne fremgangsmåten beskriver isoleringen av chromatophore granuler som brukes for å utforske de molekylære bidrag til coloration i blekkspruter. 12 lite molekyl utdrag fra hele dyr kan ofte være en lang og langtekkelig prosess. Målet her er å informere fremtidige forskere av en effektiv og lettvint protokoll for anskaffelsen av pigmenter fra nanostrukturerte granulater i blekkspruter.

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			<td height="42" style="height:42px;width:216px;">Collagenase</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.

Kromatofor dissekeres fra D. pealeii dorsal mantelen (figur 1A, 1B). Når de er fjernet, blir kromatofor lysert og renset ved hjelp av sentrifugering og vaskesykluser for å isolere ut de pigmenterte granuler (figurene 2A, 2B). Sure oppløsninger metanol (HCl-MeOH) benyttes for å ekstrahere pigment fra kornene (figur 2C), noe som ga et oppløselig pigment ekstrakt og uoppløselige, fargeløse p...

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 protokoll for utvinning av pigmenter fra nanostrukturerte granulene i blekksprut Doryteuthis pealeii kromatofor er presentert.

En metode for utvinning av 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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