Name: is my mouse pregnant? high-frequency ultrasound assessment
Uploaded: 2021-03-18T14:00:00
Description: Watch this Scientific Journal Video about Is My Mouse Pregnant? High-Frequency Ultrasound Assessment at JoVE.com

Alle trin i denne protokol følger vejledningen for pleje og brug af laboratoriedyr offentliggjort af National Institutes of Health og er blevet godkendt af Institutional Animal Care and Use Committee of New York University Grossman School of Medicine.
1. Parring af mus til tidsbestemte graviditeter
<ol><li>Par den relevante kvindelige mus (normalt en heterozygote) i et bur med den passende mandlige mus (normalt en heterozygote) til parring natten over.</li>
	<li>Adskil musene næste morgen....

<ol>
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R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+Genome+Informatics+(MGI)+is+the+international+resource+for+information+on+the+laboratory+mouse.">Mouse Genome Informatics (MGI) is the international resource for information on the laboratory mouse.</a> Methods in Molecular Biology. 1757, 141-161 (2018).</li><li>Rydell-T&#246;rm&#228;nen, K., Johnson, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+applicability+of+mouse+models+to+the+study+of+human+disease.">The applicability of mouse models to the study of human disease.</a> Methods in Molecular Biology. 1940, 3-22 (2019).</li><li>Hinton, R. B., Yutzey, K. 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V., Nagy, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Selecting+female+mice+in+estrus+and+checking+plugs.">Selecting female mice in estrus and checking plugs.</a> Cold Spring Harbor Protocols. 2016 (8), (2016).</li><li>Heyne, G. W., 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=A+simple+and+reliable+method+for+early+pregnancy+detection+in+inbred+mice.">A simple and reliable method for early pregnancy detection in inbred mice.</a> Journal of the American Association for Laboratory Animal Science. 54 (4), 368-371 (2015).</li><li>Finlay, J. B., Liu, X., Ermel, R. W., Adamson, T. 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L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Noninvasive+localization+of+nuclear+factor+of+activated+T+cells+c1-/-+mouse+embryos+by+ultrasound+biomicroscopy-Doppler+allows+genotype-phenotype+correlation.">Noninvasive localization of nuclear factor of activated T cells c1-/- mouse embryos by ultrasound biomicroscopy-Doppler allows genotype-phenotype correlation.</a> Journal of the American Society of Echocardiography. 18 (12), 1415-1421 (2005).</li><li>Kulandavelu, S., 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=Embryonic+and+neonatal+phenotyping+of+genetically+engineered+mice.">Embryonic and neonatal phenotyping of genetically engineered mice.</a> ILAR Journal. 47 (2), 103-117 (2006).</li><li>Mu, J., Slevin, J. C., Qu, D., McCormick, S., Adamson, S. 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(128), e56207 (2017).</li><li>Greco, A., 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=High+frequency+ultrasound+for+in+vivo+pregnancy+diagnosis+and+staging+of+placental+and+fetal+development+in+mice.">High frequency ultrasound for in vivo pregnancy diagnosis and staging of placental and fetal development in mice.</a> PLoS One. 8 (10), 77205 (2013).</li><li>Flores, L. E., Hildebrandt, T. B., K&#252;hl, A. A., Drews, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+detection+and+staging+of+spontaneous+embryo+resorption+by+ultrasound+biomicroscopy+in+murine+pregnancy.">Early detection and staging of spontaneous embryo resorption by ultrasound biomicroscopy in murine pregnancy.</a> Reproductive Biology and Endocrinology. 12, 38 (2014).</li><li>Norton, W. 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Det vigtigste første skridt i billeddannelsen er at identificere skeden og derefter at bestemme bifurcation af livmoderhornet til venstre og højre. Ved at følge hver livmoderhorn, imager er mindre tilbøjelige til at mis-identificere sløjfer af tarmen som livmoderen. Desuden er det vigtigt at forstå variationerne i tarmens udseende (med / uden fækalt stof) for at skelne disse fra livmoderen; lejlighedsvis, fækal "bolde" i tarmen sløjfer kan efterligne en gravid (gravid) livmoder. Selv om andre forfattere har besk...

Musen er den foretrukne pattedyrsmodel for mange menneskelige sygdomme og biologiske processer1,2,3,4. Forskning i udviklingsbiologi kræver ofte iscenesatte gravide mus for at bestemme udviklende processer på forskellige tidspunkter5,6,7,8. Desuden kræver optimal og effektiv opdræt af modelmus en vurdering af tidsbestemte graviditeter, især når efterforskere studerer virkningerne af en genmutation på udviklingen. Typisk, efterforskere mate heterozygous mus natten over, kigge efter en vaginal stik tidligt næste morgen, og håber, at en graviditet ensues9. Bestemmelse intrauterin tab starter typisk med at kontrollere et nyfødt kuld for mendelianske forhold af genotyper, derefter arbejde baglæns ved at ofre gravide mus på forskellige svangerskabsstadier og genvinde embryonerne. Efterforskere kan bestemme vægtøgning som en måling af en positiv graviditet10,11; Men, især med gensplejsede mus, kan kuldene være meget små og efterfølgende resorberes, når der er intrauterin tab, som vægtøgningen måske ikke er indlysende (især tidligt i graviditeten, ~ E6.5-8.5). En mus kan forekomme falsk gravid på grund af for eksempel en godartet abdominal tumor. I det væsentlige virker man "blind".
Ultralydbiomikroskopi i høj opløsning giver mulighed for direkte visualisering af den gravide livmoder og udvikling af museembryoner12,13,14,15,16. Selvom vi oprindeligt havde udviklet metoder til vurdering af embryonal mus hjerte-kar-fysiologi16,17, anerkendte vi nytten af denne billeddannelse modalitet for at strømline vores museavl. Specifikt måtte vi ikke længere vente med at "se", om en mus var gravid, baseret på enten den åbenlyse vægtforøgelse eller levering af et kuld; vi kunne bestemme gravid tilstand og re-mate mus hurtigt, hvis dæmningen ikke var gravid. Desuden kunne tab inden for livmoderen også let afbildes, og en tidslinje over tab kunne bestemmes uden at ofre musen (se figur 1 for en skematisk). Tid, værdifulde modelmus og midler kan således spares.

<table>
	<tbody>
		<tr>
			<td>Depilatory cream</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Ethanol, 70%</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Fur clippers</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Gauze or KimWipes</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Medical oxygen (optional)</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Medical tape</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Mouse imaging system (including anesthesia set-up and imaging platform)</td>
			<td>Fujifilm Visual Sonics</td>
			<td>Various</td>
			<td>Any system with 40 MHz center-frequency ultrasound transducer probe</td>
		</tr>
		<tr>
			<td>Razor blade (not a safety razor)</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Scale (to weigh mouse)</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Ultrasound gel</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

is my mouse pregnant? high-frequency ultrasound assessment

Musen er den foretrukne pattedyrsdyrmodel for mange menneskelige sygdomme og biologiske processer. Udviklingsbiologi kræver ofte iscenesatte gravide mus for at bestemme udviklende processer på forskellige tidspunkter. Desuden kræver optimal og effektiv opdræt af modelmus en vurdering af tidsbestemte graviditeter. Oftest parres mus natten over, og tilstedeværelsen af et vaginalt stik bestemmes; Men den positive forudsigende værdi af denne teknik er suboptimal, og man er nødt til at vente med at vide, om musen er virkelig gravid. Ultralydbiomikroskopi i høj opløsning er et effektivt værktøj til billeddannelse: 1) Om en mus er gravid; 2) Hvilken svangerskabsfase musen har nået; og 3) Om der er intrauterin tab. Ud over embryoner og fostre, skal investigator også genkende fælles artefakter i bughulen for ikke at forveksle disse for en gravid livmoder. Denne artikel indeholder en protokol til billedbehandling sammen med illustrative eksempler.

Denne protokol vil gøre det muligt for en efterforsker at afgøre trygt, om en mus er gravid, herunder i de tidlige stadier og at afgøre, om der er åbenlyse prænatal embryonale eller fostertab uden at skulle ofre den gravide dæmning. Denne protokol er især nyttig, når der opdrættes gensplejsede mus; typisk, heterozygotiske x heterozygous krydser at give homozygous afkom fører til manglende ordentlig udvikling, som forårsager prænatal dødelighed. Figur 1 viser en repræsentativ si...

Watch this Scientific Journal Video about Is My Mouse Pregnant? High-Frequency Ultrasound Assessment at JoVE.com

Is My Mouse Pregnant? High-Frequency Ultrasound Assessment

Ultralyd i høj opløsning kan hjælpe med at strømline eksperimenter, der kræver tidsbestemte gravide mus ved at bestemme tilstanden af graviditet, svangerskabsalder og graviditetstab. Præsenteret her er en protokol til at illustrere metoder til at vurdere musegraviditeter samt potentielle faldgruber (billede artefakter), der kan efterligne graviditet.

Er min mus gravid? Ultralydsvurdering med høj frekvens

The mouse is the mammalian animal model of choice for many human diseases and biological processes. Developmental biology often requires staged-pregnant ...

Ultralyd med høj opløsning kan hjælpe med at strømline eksperimenter, der kræver tidsbestemte gravide mus ved at bestemme tilstanden af graviditet, svangerskabsalder og graviditetstab. Denne protokol viser, hvordan man vurderer musegraviditeter og identificerer potentielle billedartefakter, der kan efterligne graviditet. Til at begynde med skal du identificere blæren på skærmen.Scanning kausalt fra blæren, identificere skeden. Når ultralyd transducer er placeret midt på linje og lavt på maven, blæren og skeden er synlige. Derefter scanne langsomt og glat i en kranieretning, identificere bifurcation af skeden i venstre og højre livmoder horn.Undersøg venstre og højre livmoderhorn. Op til midten af svangerskabet vil museembryoner blive placeret langs højre og venstre periferi. Efterhånden som de vokser, vil de mere distale dele af livmoderen og deres tilsvarende embryoner vende udad og dorsalt.Denne protokol er især nyttig, når der opdrættes gensplejsede mus. Typisk, heterozygotiske med heterozygotiske krydser at give homozygous afkom føre til manglende ordentlig udvikling, forårsager prænatal dødelighed. Musefostre på forskellige udviklingsstadier er vist her.Tidlige musefostre, døde embryoner eller resorberet embryoner kan ligne andre organer i maven eller afføring i tarmene, mens tarmsløjfer kan efterligne den ikke-gravide livmoder. Nogle efterforskere foretrækker at bruge en fasemonteret transducer, som vist på dette billede, men vi foretrækker den håndholdte metode til scanning, da dette giver mulighed for hurtig og effektiv billeddannelse i hele maven.

Research

JoVE Journal

Developmental Biology

Comprehensive Assessment of Germline Chemical Toxicity Using the Nematode Caenorhabditis elegans

Comprehensive Assessment of Germline Chemical Toxicity Using the Nematode Caenorhabditis elegans

Omfattende vurdering af Kimcellelinje kemiske toksicitet Brug nematoden<em&gt; Caenorhabditis elegans</em

Identifying the reproductive toxicity of the thousands of chemicals present in our environment has been one of the most tantalizing challenges in the ...

Contrast Imaging in Mouse Embryos Using High-frequency Ultrasound

Kontrast Imaging i museembryoer Brug højfrekvent ultralyd

Ultrasound contrast-enhanced imaging can convey essential quantitative information regarding tissue vascularity and perfusion and, in targeted ...

Assessment of Endothelial Cell Migration After Exposure to Toxic Chemicals

Vurdering af endotelcellemigration Efter eksponering for giftige kemikalier

Exposure to chemical substances (including alkylating chemical warfare agents like sulfur and nitrogen mustards) cause a plethora of clinical symptoms ...

Assessment of Maternal Vascular Remodeling During Pregnancy in the Mouse Uterus

Vurdering af Maternal Vascular Remodeling under graviditet i Mus livmoderen

The placenta mediates the exchange of factors such as gases and nutrients between mother and fetus and has specific demands for supply of blood from the ...

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

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 ...

Fetal Mouse Cardiovascular Imaging Using a High-frequency Ultrasound (30/45MHZ) System

Fetal Mouse Cardiovascular Imaging Using a High-frequency Ultrasound 30/45MHZ System

Føtal mus kardiovaskulære Imaging ved hjælp af en høj frekvens ultralyd (30 / 45MHZ)

Congenital heart defects (CHDs) are the most common cause of childhood morbidity and early mortality. Prenatal detection&#160;of the underlying molecular ...

High Frequency Ultrasound for the Analysis of Fetal and Placental Development In Vivo

High Frequency Ultrasound for the Analysis of Fetal and Placental Development In Vivo

Høj frekvens ultralyd til analyse af føtal og placenta udvikling In Vivo

Ultrasound imaging is a widespread method used to detect organ anomalies and tumors in human and animal tissues. The method is non-invasive, harmless, and ...

Fluorimetric Techniques for the Assessment of Sperm Membranes

Fluorimetri teknikker til vurdering af Sperm membraner

Standard spermiograms describing sperm quality are mostly based on the physiological and visual parameters, such as ejaculate volume and concentration, ...

Assessment of Ultrastructural Neuroplasticity Parameters After In Utero Transduction of the Developing Mouse Brain and Spinal Cord

Vurdering af ultrastrukturelle neuroplasticitet parametre efter i Utero transduktion af udvikle mus hjernen og rygmarven

The present study combines in utero transduction with transmission electron microscopy (TEM) aiming at a precise morphometrical analysis of ...

Assessment of Oxidative Damage in the Primary Mouse Ocular Surface Cells/Stem Cells in Response to Ultraviolet-C (UV-C) Damage

Assessment of Oxidative Damage in the Primary Mouse Ocular Surface Cells/Stem Cells in Response to Ultraviolet-C UV-C Damage

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The ocular surface is subjected to regular wear and tear due to various environmental factors. Exposure to UV-C radiation constitutes an occupational ...