Name: optimization of the retinal vein occlusion mouse model to limit variability
Uploaded: 2021-08-06T15:00:00
Description: Watch this Scientific Journal Video about Optimization of the Retinal Vein Occlusion Mouse Model to Limit Variability at JoVE.com

<div>Dette arbejde blev støttet af National Science Foundation Graduate Research Fellowship Program (NSF-GRFP) DGE - 1644869 (til CCO), National Eye Institute (NEI) 5T32EY013933 (til AMP) og National Institute on Aging (NIA) R21AG063012 (til CMT).</div>

Denne protokol følger erklæringen fra Association for Research in Vision and Ophthalmology (ARVO) om brug af dyr i oftalmisk og synsforskning. Gnaverforsøg blev godkendt og overvåget af Institutional Animal Care and Use Committee (IACUC) ved Columbia University.
BEMÆRK: Alle forsøg brugte to måneder gamle hanmus, der vejede ca. 20 g.
1. Forberedelse og administration af tamoxifen til inducerbar genetisk ablation af floxede gener
BEM?...

<ol>
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Musens RVO-model giver en mulighed for yderligere at forstå RVO-patologi og teste potentielle terapi. Mens musens RVO-model er meget udbredt i marken, er der behov for en aktuel detaljeret protokol for modellen, der adresserer dens variabilitet og beskriver optimeringen af modellen. Her giver vi en guide med eksempler fra erfaringer på, hvad der kan ændres for at få de mest konsistente resultater på tværs af en kohorte af forsøgsdyr og give pålidelige data.
De to vigtigste elementer i ...

Retinal veneokklusion (RVO) er en almindelig retinal vaskulær sygdom, der ramte ca. 28 millioner mennesker verden over i 20151. RVO fører til synstab og tab hos voksne og ældre i den erhvervsaktive alder, hvilket repræsenterer en igangværende synstruende sygdom, der anslås at stige i løbet af det nærmeste årti. Nogle af de forskellige patologier af RVO omfatter hypoxisk-iskæmisk skade, retinal ødem, inflammation og neuronalt tab2. I øjeblikket er den første behandlingslinje for denne lidelse gennem administration af vaskulære endotelvækstfaktorhæmmere (VEGF). Mens anti-VEGF-behandling har hjulpet med at forbedre nethindeødem, står mange patienter stadig over for synsfald3. For yderligere at forstå patofysiologien af denne sygdom og for at teste potentielle nye behandlingslinjer er der behov for at udgøre en funktionel og detaljeret RVO musemodelprotokol for forskellige musestammer.
Musemodeller er blevet udviklet til implementering af den samme laserenhed, der anvendes til menneskelige patienter, parret med et billeddannelsessystem, der er skaleret til den korrekte størrelse for en mus. Denne musemodel af RVO blev først rapporteret i 2007 4 og yderligere etableret af Ebneter og andre 4,5. Til sidst blev modellen optimeret af Fuma et al. til at replikere vigtige kliniske manifestationer af RVO såsom nethindeødem6. Siden modellen først blev rapporteret, har mange undersøgelser anvendt den ved hjælp af administration af et fotosensibiliserende farvestof efterfulgt af fotokoagulation af større retinale vener med en laser. Imidlertid varierer mængden og typen af farvestoffet, der administreres, laserkraft og eksponeringstid betydeligt på tværs af undersøgelser, der har brugt denne metode. Disse forskelle kan ofte føre til øget variation i modellen, hvilket gør den vanskelig at replikere. Til dato er der ingen offentliggjorte undersøgelser med specifikke detaljer om potentielle muligheder for optimering.
Denne rapport præsenterer en detaljeret metodologi for RVO musemodellen i C57BL/6J stammen og en tamoxifen-inducerbar endotel-caspase-9 knockout (iEC Casp9KO) stamme med en C57BL/6J baggrund og af relevans for RVO patologi som referencestamme for en genetisk modificeret mus. En tidligere undersøgelse havde vist, at ikke-apoptotisk aktivering af endotel-caspase-9 anstifter retinal ødem og fremmer neuronal død8. Erfaring med at bruge denne stamme hjalp med at bestemme og give indsigt i potentielle ændringer for at skræddersy RVO musemodellen, som kan anvendes til andre genetisk modificerede stammer.

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			<td>Alcon</td>
			<td>00658 06401</td>
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			<td>Ketamine Hydrochloride</td>
			<td>Henry Schein</td>
			<td>NDC: 11695-0702-1</td>
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			<td>Lasercheck</td>
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			<td>Phoneix Micron IV with Meridian,&#160; StreamPix, and OCT modules</td>
			<td>Phoenix Technology Group</td>
			<td></td>
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			<td>Proparacaine Hydrochloride</td>
			<td>Akorn</td>
			<td>NDC: 17478-263-12</td>
			<td>keep at 4 &#176;C</td>
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			<td>Refresh</td>
			<td>Allergan</td>
			<td>94170</td>
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			<td>Rose Bengal</td>
			<td>Sigma-Aldrich</td>
			<td>330000-5G</td>
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			<td>Tamoxifen</td>
			<td>Sigma-Aldrich</td>
			<td>T5648-5G</td>
			<td>light-sensitive</td>
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			<td>Tropicamide</td>
			<td>Akorn</td>
			<td>NDC: 174478-102-12</td>
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			<td>Xylazine</td>
			<td>Akorn</td>
			<td>NDCL 59399-110-20</td>
			<td></td>
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optimization of the retinal vein occlusion mouse model to limit variability

Musemodeller af retinal veneokklusion (RVO) bruges ofte i oftalmologi til at studere hypoxisk-iskæmisk skade i nethinden i nethinden . I denne rapport gives en detaljeret metode, der påpeger kritiske trin, med anbefalinger til optimering for at opnå konsekvent vellykkede okklusionshastigheder på tværs af forskellige genetisk modificerede musestammer. RVO musemodellen består primært af intravenøs administration af et fotosensibiliserende farvestof efterfulgt af laserfotokoagulation ved hjælp af et retinal billedmikroskop fastgjort til en oftalmisk guidet laser. Tre variabler blev identificeret som determinanter for okklusionskonsistens. Ved at justere ventetiden efter administration af rosenbengalen og afbalancere baseline og eksperimentel laseroutput kan variabiliteten på tværs af eksperimenter begrænses, og der opnås en højere succesrate for okklusioner. Denne metode kan bruges til at studere retinale sygdomme, der er karakteriseret ved retinal ødem og hypoxisk-iskæmisk skade. Derudover, da denne model inducerer vaskulær skade, kan den også anvendes til at studere neurovaskulaturen, neuronal død og betændelse.

RVO musemodellen sigter mod med succes at opnå okklusioner i retinale vener, hvilket fører til hypoxisk-iskæmisk skade, nedbrydning af blodets retinale barriere, neuronal død og retinal ødem8. Figur 1 viser en tidslinje med trin for at sikre reproducerbarhed, et skema over det eksperimentelle design og skitserer trin, der kan optimeres yderligere afhængigt af de eksperimentelle spørgsmål. De tre vigtigste trin, der kan ændres, er ventetiden efter rosenbengala...

Watch this Scientific Journal Video about Optimization of the Retinal Vein Occlusion Mouse Model to Limit Variability at JoVE.com

Optimization of the Retinal Vein Occlusion Mouse Model to Limit Variability

Her beskriver vi en optimeret protokol til retinal veneokklusion ved hjælp af rose bengal og et laserstyret retinal billeddannelsesmikroskopsystem med anbefalinger for at maksimere dets reproducerbarhed i genetisk modificerede stammer.

Optimering af musemodellen med retinal veneokklusion for at begrænse variabilitet

Mouse models of retinal vein occlusion (RVO) are often used in ophthalmology to study hypoxic-ischemic injury in the neural retina. In this report, a ...

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