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

本议定书的所有步骤均遵循美国国家卫生研究院出版的《实验室动物护理和使用指南》，并已得到纽约大学格罗斯曼医学院机构动物护理和使用委员会的批准。
1. 老鼠为早产而交配
<ol><li>将适当的雌性小鼠（通常是异质鼠）与适当的雄性小鼠（通常是异质鼠）配对，进行夜间交配。</li>
	<li>第二天早上把老鼠分开。或者，继续与雌性小鼠和雄性小鼠交配，从而增加怀孕的机?...

<ol>
	<li>Bogue, M. 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=Mouse+Phenome+Database:+an+integrative+database+and+analysis+suite+for+curated+empirical+phenotype+data+from+laboratory+mice.">Mouse Phenome Database: an integrative database and analysis suite for curated empirical phenotype data from laboratory mice.</a> Nucleic Acids Research. 46, 843-850 (2018).</li><li>Ito, R., Takahashi, T., Ito, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Humanized+mouse+models:+Application+to+human+diseases.">Humanized mouse models: Application to human diseases.</a> Journal of Cellular Physiology. 233 (5), 3723-3728 (2018).</li><li>Law, M., Shaw, D. 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L., 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=Formation+of+the+embryonic+head+in+the+mouse:+attributes+of+a+gene+regulatory+network.">Formation of the embryonic head in the mouse: attributes of a gene regulatory network.</a> Current Topics in Developmental Biology. 117, 497-521 (2016).</li><li>Palis, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hematopoietic+stem+cell-independent+hematopoiesis:+emergence+of+erythroid,+megakaryocyte,+and+myeloid+potential+in+the+mammalian+embryo.">Hematopoietic stem cell-independent hematopoiesis: emergence of erythroid, megakaryocyte, and myeloid potential in the mammalian embryo.</a> FEBS Letters. 590 (22), 3965-3974 (2016).</li><li>Behringer, R., Gertsenstein, M., Nagy, K. 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. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Maternal+weight+gain+as+a+predictor+of+litter+size+in+Swiss+Webster,+C57BL/6J,+and+BALB/cJ+mice.">Maternal weight gain as a predictor of litter size in Swiss Webster, C57BL/6J, and BALB/cJ mice.</a> Journal of the American Association for Laboratory Animal Science. 54 (6), 694-699 (2015).</li><li>Zhou, Y. Q., 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=Applications+for+multifrequency+ultrasound+biomicroscopy+in+mice+from+implantation+to+adulthood.">Applications for multifrequency ultrasound biomicroscopy in mice from implantation to adulthood.</a> Physiological Genomics. 10 (2), 113-126 (2002).</li><li>Ji, R. P., Phoon, C. K. 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. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+quantification+of+embryonic+and+placental+growth+during+gestation+in+mice+using+micro-ultrasound.">In vivo quantification of embryonic and placental growth during gestation in mice using micro-ultrasound.</a> Reproductive Biology and Endocrinology. 6, 34 (2008).</li><li>Phoon, C. K. L., Turnbull, D. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiovascular+imaging+in+mice.">Cardiovascular imaging in mice.</a> Current Protocols in Mouse Biology. 6 (1), 15-38 (2016).</li><li>Phoon, C. K. L., Turnbull, D. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ultrasound+biomicroscopy-Doppler+in+mouse+cardiovascular+development.">Ultrasound biomicroscopy-Doppler in mouse cardiovascular development.</a> Physiological Genomics. 14 (1), 3-15 (2003).</li><li>Peavey, M. 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=A+novel+use+of+three-dimensional+high-frequency+ultrasonography+for+early+pregnancy+characterization+in+the+mouse.">A novel use of three-dimensional high-frequency ultrasonography for early pregnancy characterization in the mouse.</a> Journal of Visualized Experiments. (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. B., 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=Refinements+for+embryo+implantation+surgery+in+the+mouse:+comparison+of+injectable+and+inhalant+anesthesias+-+tribromoethanol,+ketamine+and+isoflurane+-+on+pregnancy+and+pup+survival.">Refinements for embryo implantation surgery in the mouse: comparison of injectable and inhalant anesthesias - tribromoethanol, ketamine and isoflurane - on pregnancy and pup survival.</a> Laboratory Animal. 50 (5), 335-343 (2016).</li><li>Thaete, L. G., Levin, S. I., Dudley, A. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impact+of+anaesthetics+and+analgesics+on+fetal+growth+in+the+mouse.">Impact of anaesthetics and analgesics on fetal growth in the mouse.</a> Laboratory Animal. 47 (3), 175-183 (2013).</li><li>Phoon, C. K. 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成像中最重要的第一步是识别阴道，然后确定左右两侧子宫角的分叉。通过跟随每个子宫喇叭，成像仪不太可能错误地识别肠道的循环作为子宫。此外，了解肠道外观的变化（有/无粪便物质）对于区分这些和子宫非常重要：偶尔，排便环中的粪便"球"可能模仿大肠（怀孕）子宫。虽然其他作者描述了诊断怀孕和分期小鼠胚胎发育17，18，19包括检测重新吸收的胚胎<sup class=...

小鼠是许多人类疾病和生物过程1，2，3，4的首选哺乳动物模型。发育生物学研究往往要求分阶段怀孕的小鼠确定进化过程在不同的时间点5，6，7，8。此外，模型小鼠的最佳和有效繁殖需要评估定时怀孕，尤其是当研究人员正在研究基因突变对发育的影响时。通常，研究人员在一夜之间与异质小鼠交配，第二天一早寻找阴道塞子，并希望怀孕后9日。确定宫内损失通常从检查新生儿的粪便中是否与门代利人基因型的比例开始，然后通过在不同的妊娠阶段牺牲怀孕的小鼠和恢复胚胎来向后工作。研究者可以确定体重增加是阳性妊娠10、11的指标：然而，特别是与基因工程小鼠，垃圾可能非常小，随后重新吸收时，有宫内损失，由于体重增加可能并不明显（特别是在怀孕早期，~E6.5-8.5）。例如，由于良性腹部肿瘤，小鼠可能出现虚假怀孕。从本质上讲，一个人的作品是"盲目的"。
高分辨率超声波生物显微镜允许直接可视化格拉维德子宫和开发小鼠胚胎12，13，14，15，16。虽然我们最初已经开发出评估胚胎小鼠心血管生理学的方法16，17，我们认识到这种成像方式的效用，以简化我们的小鼠繁殖。具体来说，我们不再需要等待"看"老鼠是否怀孕，基于明显的体重增加或运送垃圾：如果大坝没有怀孕，我们可以确定鼠鰿的状态，并迅速重新交配小鼠。此外，宫内损失也可以很容易地被映像，在不牺牲鼠标的情况下可以确定损失的时间表（见图1的示意图）。因此，可以节省时间、有价值的模型鼠标和资金。

<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

小鼠是许多人类疾病和生物过程的首选哺乳动物动物模型。发育生物学通常需要分阶段怀孕的小鼠来确定不同时间点的进化过程。此外，模型小鼠的最佳和高效繁殖需要对定时怀孕进行评估。最常见的情况是，小鼠在一夜之间交用，阴道塞的存在被确定：然而，这种技术的正预测值是次优的，人们需要等待知道老鼠是否真的怀孕了。高分辨率超声波生物显微镜是一种有效有效的成像工具：1） 小鼠是否怀孕：2） 鼠标达到的妊娠阶段：和 3） 是否有宫内损失。除了胚胎和胎儿外，研究者还必须识别腹腔中的常见神器，以免误以为这些是子宫。本文提供了成像协议以及说明性示例。

该协议将允许调查员自信地确定小鼠是否怀孕，包括在早期阶段，并确定是否有明显的产前胚胎或胎儿损失，而无需牺牲怀孕的水坝。这种协议在培育转基因小鼠时特别有用：通常，异质x异质交叉产生同源后代会导致正常发育失败，从而导致产前杀伤力。图1描绘了胚胎在妊娠中期逐渐死亡，然后被重新吸收的代表性情况。图2显示了如何通过通过阴道?...

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

高分辨率超声波可以通过确定怀孕状态、妊娠年龄和怀孕损失来帮助简化需要定时怀孕小鼠的实验。这里介绍的是一个协议，以说明评估小鼠怀孕的方法，以及可能模仿怀孕的潜在陷阱（图像文物）。

我的老鼠怀孕了吗？高频超声波评估

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

高分辨率超声波可以通过确定怀孕状态、妊娠年龄和怀孕损失，帮助简化需要定时怀孕小鼠的实验。该协议演示了如何评估小鼠怀孕和识别可能模仿怀孕的潜在图像人工制品。首先，识别屏幕上的膀胱。从膀胱中扫描，识别阴道。当超声波传感器位于腹部的中线和低位时，膀胱和阴道是可见的。然后缓慢而平稳地向颅面方向扫描，确定阴道向左右子宫角的分叉。调查左右子宫角。在妊娠中期，小鼠胚胎将沿着左右外围放置。随着它们的生长，子宫及其相应胚胎的分离部分会向外和向后转动。这个协议在培育转基因小鼠时特别有用。通常，异质性与异质交叉产生同源后代会导致正常发育失败，导致产前杀伤力。此处显示了处于不同发育阶段的鼠标胚胎。早期小鼠胚胎、死胚胎或重新吸收的胚胎可能类似于腹部或肠道粪便中的其他器官，而肠道循环可能模仿非腹腔。一些研究者喜欢使用舞台安装的传感器，如此图像中显示的，但我们更喜欢手持式扫描方法，因为这允许快速高效地成像整个腹部。

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

生殖系化学毒性的利用线虫综合评估<em&gt;秀丽隐杆线虫</em

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

科研

发育生物学

Contrast Imaging in Mouse Embryos Using High-frequency Ultrasound

对比成像的小鼠胚胎使用高频超声

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

内皮细胞迁移评估暴露于有毒化学品后，

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

产妇血管重构的评估在怀孕期间在小鼠子宫

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

三维高频超声对小鼠早期妊娠特征的新应用

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

用高频超声 (30/45 mhz) 系统对胎儿小鼠进行心血管成像

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

高频超声在胎儿和胎盘发育分析中的影响

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

精子膜评价的荧光技术

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

小鼠脑脊髓子宫转导后超微结构神经可塑性参数的评价

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

评估原小鼠眼部表面细胞/干细胞对紫外线-C （UV-C） 损伤的反应

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