Name: a novel use of three-dimensional high-frequency ultrasonography for early pregnancy characterization in the mouse
Uploaded: 2017-10-24T13:00:00
Description: Watch this Scientific Journal Video about A Novel Use of Three-dimensional High-frequency Ultrasonography for Early Pregnancy Characterization in the Mouse at JoVE.com

Vi værdsætter i høj grad bistand fra Rong Zhao, Jie Li og Yan Ying.

disse undersøgelser blev gennemført i overensstemmelse med retningslinjer for pleje og anvendelse af forsøgsdyr udgivet af National Institutes of Health og animalske protokoller godkendt af institutionelle Animal Care og brug udvalg (IACUC) af Baylor College of Medicine under protokollen antal AN-4203.
 1. forberedelse af den gravide mus for ultralyd <ol><li>timet parring <ol><li>sted dæmningen med en dokumenteret frugtbar mandlige mus overnight begyndelsen efter 1700. ...

<ol>
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C., 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=Three-Dimensional+High-Frequency+Ultrasonography+for+Early+Detection+and+Characterization+of+Embryo+Implantation+Site+Development+in+the+Mouse.">Three-Dimensional High-Frequency Ultrasonography for Early Detection and Characterization of Embryo Implantation Site Development in the Mouse.</a> PLoS One. 12 (1), e0169312 (2017).</li><li>Song, H., 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=Cytosolic+phospholipase+A2alpha+is+crucial+[correction+of+A2alpha+deficiency+is+crucial]+for+'on-time'+embryo+implantation+that+directs+subsequent+development.">Cytosolic phospholipase A2alpha is crucial [correction of A2alpha deficiency is crucial] for 'on-time' embryo implantation that directs subsequent development.</a> Development. 129 (12), 2879-2889 (2002).</li><li>Nallasamy, S., Li, Q., Bagchi, M. 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Denne roman anvendelse af 3-D HFUS billeddannelse demonstrerer evne til med held at opdage, visualisere og karakterisere embryo-implantation websteder i den tidlige murine graviditet i en ikke-invasiv måde. Teknologien tilbyder en betydelig forbedring over nuværende metoder, der er afhængige af afbrydelsen af graviditeter for grov væv og histopatologisk karakterisering. Dog skal det bemærkes, at histologiske metoder ville stadig blive betragtet som mere optimal når karakterisering på en mere forstørret og mere ce...

Tilbagevendende tidlig graviditet tab er en af de mest almindelige komplikationer efter undfangelsen og rammer cirka 1% af par forsøger at blive gravide1,2. De underliggende mekanismer i tidlig graviditet tab er forskellige: fra iboende embryonale abnormiteter og maternel co-morbiditet defekter i endometrie modtagelighed1,3,4. På grund af deres genetiske sporbarhed, har musemodeller været bredt udnyttet for undersøgelser af tidlig embryo implantation og graviditet. Desuden, den korte svangerskabsuge tid musen og evnen til at udføre store undersøgelser har sikret den voksende nytte af mus i håndteringen af centrale kliniske spørgsmål i menneskelige forplantningsmedicin5. Som sagt, det store flertal af murine eksperimentelle design stadig kræve adskillige dæmninger til aflives på sekventiel svangerskabsuge dage at kvantificere og analysere implantation site placering, antal, størrelse og afstand mønstre under graviditet6, 7,8, derved udelukkes longitudinelle studier på samme dyr.
I klinikken er ultralyd en pålidelig og uvurderligt redskab til at overvåge menneskelige føtal levedygtighed og udvikling i en ikke-invasiv måde9,10,11. Mere nylig, høj frekvens ultralyd (HFUS) er begyndt at finde begrænset anvendelse i mus som en metode til overvågning af føtal levedygtighed og væksten under graviditeten12,13,14. De seneste teknologiske fremskridt i ultrasound imaging har tilladt anvendelsen af tre-dimensionelle (3-D) data for visuel genopbygning af dyrs organer og efterfølgende overvågning af patologier15,16, 17. Brug af denne avancerede imaging-teknologi har markant forbedret beføjelse til at opdage mindre volumen udsving, at reducere Inter animalske variabilitet og overvåge udviklingen af en patologi eller effekten af en terapeutisk intervention17. Mens den primære nytte af denne teknologi har været at overvåge malignitet progression i oncomouse modeller15,16, er 3-D HFUS billeddannelse først for nylig blevet brugt til at kvantificere og overvåge aktiv vækst af fosteret implantation og fostrets udvikling i mus livmoderen18.
Her, viser vi hvordan man udfører HFUS imaging for at fremstille 2D- og 3D-data for at generere rekonstruktioner af tidlige gravide mus livmoderen. Vi påvise nytten af denne nye metode til at opdage disse tidlige embryonale implantation begivenheder uden behov for graviditet opsigelse, giver forskerne til at indsamle data i en ikke-invasiv måde.

<table>
	<tbody>
		<tr>
			<td>VisualSonics Vevo 2100 Ultrasound Imaging Platform/Machine</td>
			<td>VisualSonics, inc.</td>
			<td>VS-11945</td>
			<td></td>
		</tr>
		<tr>
			<td>Vevo Imaging Station</td>
			<td>VisualSonics, inc.</td>
			<td>SA-11982</td>
			<td></td>
		</tr>
		<tr>
			<td>Aquasonic 100 Ultrasound Transmission Gel</td>
			<td>Parker</td>
			<td>#SKU PLI 01-08</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane (IsoThesia) 100mL bottle</td>
			<td>Henry Shein</td>
			<td>#29404</td>
			<td></td>
		</tr>
		<tr>
			<td>PuraLubenAnimal Ophthalmic Ointment</td>
			<td>Dechra</td>
			<td>#12920060</td>
			<td></td>
		</tr>
	</tbody>
</table>

a novel use of three-dimensional high-frequency ultrasonography for early pregnancy characterization in the mouse

Høj-frekvens ultralyd (HFUS) er en fælles metode til ikke-invasivt overvåge real-time udvikling af menneskelige fosteret i utero. Musen bruges rutinemæssigt som en i vivo model til at studere embryo implantation og graviditet progression. Desværre, sådan murine undersøgelser kræver graviditet afbrydelse at aktivere opfølgende fænotypiske analyser. For at løse dette problem, brugte vi tre-dimensionelle (3-D) genopbygning af HFUS imaging data for tidlig påvisning og karakterisering af murine embryo implantation sites og deres individuelle udviklingsmæssige progression in utero. Ved at kombinere HFUS imaging med 3D-genopbygning og modellering, var vi i stand til nøjagtigt kvantificere embryo implantation site nummer samt overvåge udviklingsmæssige progression i gravide C57BL6J/129S mus fra 5.5 dage post coitus (d.p.c.) gennem til 9,5 d.p.c. med brugen af en transducer. Målinger inkluderet: antal, placering og volumen af implantation websteder samt Inter implantation websted afstand; embryo levedygtighed blev vurderet af hjertets Aktivitetsovervågning. I den umiddelbare efter implantation periode (5,5 til 8,5 d.p.c.), 3D-genopbygning af fælderne livmoderen i både mesh og solid overlay format aktiveret visuel repræsentation af de udvikle graviditeter inden for hver uterin horn. Som gensplejsede mus fortsat kan anvendes til at karakterisere kvindelige reproduktive fænotyper afledt af uterin dysfunktion, giver denne metode en ny tilgang for at opdage, kvantificere og karakterisere tidlig implantation begivenheder i vivo. Denne roman anvendelse af 3-D HFUS billeddannelse demonstrerer evne til med held at opdage, visualisere og karakterisere embryo-implantation websteder i den tidlige murine graviditet i en ikke-invasiv måde. Teknologien tilbyder en betydelig forbedring over nuværende metoder, der er afhængige af afbrydelsen af graviditeter for grov væv og histopatologisk karakterisering. Her bruger vi en video og tekst format til at beskrive hvordan man med held udføre ultralyd af tidlige murine graviditet til at generere pålidelig og reproducerbar data med genopbygningen af den uterine form i mesh og solid 3D-billeder.

Som vist i figur 1, kan høj frekvens ultralyd afsløre implantation site udvikling begynder så tidligt som 5,5 d.p.c tid. Brug den lettere hyperekkoisk giver decidualized endometriet som markør af implantation websteder på 6,5 d.p.c antallet af implantation websteder og afstanden mellem disse websteder til kvantificeres. Som graviditeten er skrider frem til 7,5 d.p.c. mørkere hypoechoic svangerskabsuge sac og føtal pole også let kan identificeres.
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Watch this Scientific Journal Video about A Novel Use of Three-dimensional High-frequency Ultrasonography for Early Pregnancy Characterization in the Mouse at JoVE.com

A Novel Use of Three-dimensional High-frequency Ultrasonography for Early Pregnancy Characterization in the Mouse

En roman brug af tre-dimensionelle høj frekvens ultralyd for tidlig graviditet karakterisering i mus

Mus er meget udbredt at studere svangerskabsuge biologi. Men graviditet opsigelse kræves for sådanne undersøgelser, som er til hinder for langsgående undersøgelser og nødvendiggør brug af et stort antal dyr. Derfor, vi beskriver en non-invasiv teknik, høj frekvens ultralyd for tidlig opdagelse og overvågning af efter implantation begivenheder i den gravide mus.

High-frequency ultrasonography (HFUS) is a common method to non-invasively monitor the real-time development of the human fetus in utero. The mouse is ...

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