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

我们非常感谢蓉赵、洁丽和严营的帮助。

这些研究是按照由机构动物保育和使用委员会 (IACUC) 批准的国家卫生和动物议定书研究所出版的《爱护和使用实验室动物指南》进行的。贝勒医学院根据协议编号 AN-4203. 
 1. 用于超声的孕鼠的制备 <ol> <li> 定时交配 <ol> 在1700年后的一夜之间将大坝与一只被证明是可育的雄性老鼠放在一起。分开的水坝和男性由0700小时 (h), 不管阴道堵塞是否存在, 以确保准确的?...

<ol>
	<li>Rai, R., Regan, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recurrent+miscarriage.">Recurrent miscarriage.</a> Lancet. 368 (9535), 601-611 (2006).</li><li>Sugiura-Ogasawara, M., Ozaki, Y., Suzumori, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Management+of+recurrent+miscarriage.">Management of recurrent miscarriage.</a> J Obstet Gynaecol Res. 40 (5), 1174-1179 (2014).</li><li>Kutteh, W. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Novel+strategies+for+the+management+of+recurrent+pregnancy+loss.">Novel strategies for the management of recurrent pregnancy loss.</a> Semin Reprod Med. 33 (3), 161-168 (2015).</li><li>Page, J. M., Silver, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+Causes+of+Recurrent+Pregnancy+Loss.">Genetic Causes of Recurrent Pregnancy Loss.</a> Clin Obstet Gynecol. 59 (3), 498-508 (2016).</li><li>Zhang, J., Croy, B. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Using+ultrasonography+to+define+fetal-maternal+relationships:+moving+from+humans+to+mice.">Using ultrasonography to define fetal-maternal relationships: moving from humans to mice.</a> Comp Med. 59 (6), 527-533 (2009).</li><li>Li, S. J., 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=Differential+regulation+of+receptivity+in+two+uterine+horns+of+a+recipient+mouse+following+asynchronous+embryo+transfer.">Differential regulation of receptivity in two uterine horns of a recipient mouse following asynchronous embryo transfer.</a> Sci Rep. 5, 15897 (2015).</li><li>Ding, Y. 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=5-aza-2'-deoxycytidine+leads+to+reduced+embryo+implantation+and+reduced+expression+of+DNA+methyltransferases+and+essential+endometrial+genes.">5-aza-2'-deoxycytidine leads to reduced embryo implantation and reduced expression of DNA methyltransferases and essential endometrial genes.</a> PLoS One. 7 (9), e45364 (2012).</li><li>Kusakabe, K., Naka, M., Ito, Y., Eid, N., Otsuki, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulation+of+natural-killer+cell+cytotoxicity+and+enhancement+of+complement+factors+in+the+spontaneously+aborted+mouse+placenta.">Regulation of natural-killer cell cytotoxicity and enhancement of complement factors in the spontaneously aborted mouse placenta.</a> Fertil Steril. 90 (4 Suppl), 1451-1459 (2008).</li><li>Demianczuk, N. N., 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=The+use+of+first+trimester+ultrasound.">The use of first trimester ultrasound.</a> J Obstet Gynaecol Can. 25 (10), 864-875 (2003).</li><li>Thompson, H. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+the+obstetric+and+gynecologic+patient+by+the+use+of+diagnostic+ultrasound.">Evaluation of the obstetric and gynecologic patient by the use of diagnostic ultrasound.</a> Clin Obstet Gynecol. 17 (4), 1-25 (1974).</li><li>Unterscheider, J., 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=Definition+and+management+of+fetal+growth+restriction:+a+survey+of+contemporary+attitudes.">Definition and management of fetal growth restriction: a survey of contemporary attitudes.</a> Eur J Obstet Gynecol Reprod Biol. 174, 41-45 (2014).</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), e77205 (2013).</li><li>Flores, L. E., Hildebrandt, T. B., Kuhl, 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> Reprod Biol Endocrinol. 12, 38 (2014).</li><li>Nguyen, T. M., 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=Estimation+of+mouse+fetal+weight+by+ultrasonography:+application+from+clinic+to+laboratory.">Estimation of mouse fetal weight by ultrasonography: application from clinic to laboratory.</a> Lab Anim. 46 (3), 225-230 (2012).</li><li>Singh, 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=Quantitative+volumetric+imaging+of+normal,+neoplastic+and+hyperplastic+mouse+prostate+using+ultrasound.">Quantitative volumetric imaging of normal, neoplastic and hyperplastic mouse prostate using ultrasound.</a> BMC Urol. 15, 97 (2015).</li><li>Liu, 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=Ultrasound-mediated+destruction+of+paclitaxel+and+oxygen+loaded+lipid+microbubbles+for+combination+therapy+in+ovarian+cancer+xenografts.">Ultrasound-mediated destruction of paclitaxel and oxygen loaded lipid microbubbles for combination therapy in ovarian cancer xenografts.</a> Cancer Lett. 361 (1), 147-154 (2015).</li><li>Ni, J., 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=Monitoring+Prostate+Tumor+Growth+in+an+Orthotopic+Mouse+Model+Using+Three-Dimensional+Ultrasound+Imaging+Technique.">Monitoring Prostate Tumor Growth in an Orthotopic Mouse Model Using Three-Dimensional Ultrasound Imaging Technique.</a> Transl Oncol. 9 (1), 41-45 (2016).</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=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. K., Bagchi, I. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Msx+homeobox+genes+critically+regulate+embryo+implantation+by+controlling+paracrine+signaling+between+uterine+stroma+and+epithelium.">Msx homeobox genes critically regulate embryo implantation by controlling paracrine signaling between uterine stroma and epithelium.</a> PLoS Genet. 8 (2), e1002500 (2012).</li><li>Hirate, Y., 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+Sox17+haploinsufficiency+leads+to+female+subfertility+due+to+impaired+implantation.">Mouse Sox17 haploinsufficiency leads to female subfertility due to impaired implantation.</a> Sci Rep. 6, 24171 (2016).</li><li>Wang, T. 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=Dysregulated+LIF-STAT3+pathway+is+responsible+for+impaired+embryo+implantation+in+a+Streptozotocin-induced+diabetic+mouse+model.">Dysregulated LIF-STAT3 pathway is responsible for impaired embryo implantation in a Streptozotocin-induced diabetic mouse model.</a> Biol Open. 4 (7), 893-902 (2015).</li><li>Ji, R. P., 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=Onset+of+cardiac+function+during+early+mouse+embryogenesis+coincides+with+entry+of+primitive+erythroblasts+into+the+embryo+proper.">Onset of cardiac function during early mouse embryogenesis coincides with entry of primitive erythroblasts into the embryo proper.</a> Circ Res. 92 (2), 133-135 (2003).</li><li>Srinivasan, 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=Noninvasive,+in+utero+imaging+of+mouse+embryonic+heart+development+with+40-MHz+echocardiography.">Noninvasive, in utero imaging of mouse embryonic heart development with 40-MHz echocardiography.</a> Circulation. 98 (9), 912-918 (1998).</li><li>Franco, N. H., Olsson, I. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Scientists+and+the+3Rs:+attitudes+to+animal+use+in+biomedical+research+and+the+effect+of+mandatory+training+in+laboratory+animal+science.">Scientists and the 3Rs: attitudes to animal use in biomedical research and the effect of mandatory training in laboratory animal science.</a> Lab Anim. 48 (1), 50-60 (2014).</li><li>Pratap, K., Singh, V. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+training+course+on+laboratory+animal+science:+an+initiative+to+implement+the+Three+Rs+of+animal+research+in+India.">A training course on laboratory animal science: an initiative to implement the Three Rs of animal research in India.</a> Altern Lab Anim. 44 (1), 21-41 (2016).</li><li>Landi, M. S., Shriver, A. J., Mueller, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Consideration+and+checkboxes:+incorporating+ethics+and+science+into+the+3Rs.">Consideration and checkboxes: incorporating ethics and science into the 3Rs.</a> J Am Assoc Lab Anim Sci. 54 (2), 224-230 (2015).</li></ol>

这项新的使用3维高频成像显示的能力, 成功地检测, 可视化, 并表征胚胎植入点在早期小鼠怀孕的非侵入性的方式。该技术提供了明显的改善比目前的方法, 这依赖于中断妊娠的总组织和病理特征。然而, 应该指出的是, 组织学方法将仍然被认为是更优化, 当表征在一个更放大和更多的细胞水平是需要的, 或当基因和蛋白质分析是必需的。随着越来越多的新的鼠标模型显示植入缺陷和早期怀孕的损失<s...

复发性早孕损失是一个最常见的并发症后受孕和影响约1% 的夫妇试图受孕1,2。早期妊娠损失的基本机制多种多样: 从内在的胚胎异常和母体并存到子宫内膜容受性的缺陷1,3,4。由于其遗传可, 小鼠模型已被广泛应用于早期胚胎着床和妊娠的研究。此外, 鼠标的短孕龄和执行 large-scale 研究的能力, 确保了在解决人类生殖医学的关键临床问题5中, 老鼠的日益增加的效用。尽管如此, 绝大多数的小鼠实验设计仍然要求在妊娠天数内对大量的大坝进行安乐死, 以量化和分析怀孕期间的植入地点、数量、大小和间隔模式6, 7,8, 从而排除对同一动物的纵向研究。
在临床中, 超声是一个可靠的和无创的工具, 以监测人的胎儿生存能力和发展的非侵入性的方式9,10,11。最近, 高频超声 (高频) 已经开始发现有限的应用在鼠标作为一种方法, 监测胎儿的生存能力和生长在怀孕期间12,13,14。最近的技术进步在超声成像允许应用三维 (3) 数据为视觉重建动物器官和随后监视病症15,16, 17。使用这种先进的成像技术显著提高了检测小体积波动的能力, 减少 inter-animal 的变异性, 并监测病理的进展或治疗干预的效果17。虽然这项技术的主要效用是监测恶性进展的 oncomouse 模型15,16, 3 维高频成像最近才被用于定量和监测胚胎着床的主动增长和胎儿发育在小鼠子宫18。
在这里, 我们演示如何执行高频成像, 以产生2维和3维数据, 以生成早期怀孕的小鼠子宫的重建。我们证明了这种新方法的效用, 以检测这些早期胚胎植入事件不需要终止妊娠, 使研究人员收集数据的非侵入性的方式。

<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

高频超声 (高频) 是性监测胎儿子宫内real-time 发育的常用方法。小鼠通常被用作一个体内模型来研究胚胎着床和妊娠进展。不幸的是, 这种小鼠的研究需要怀孕中断, 以使随访表型分析。为了解决这个问题, 我们使用了三维 (3-D) 重建高频成像数据, 早期发现和表征小鼠胚胎着床地点和各自的发展进程在子宫内。结合高频成像与3维重建和建模, 我们能够准确地量化胚胎植入点的数量, 并监测发育进展的怀孕 C57BL6J/129S 小鼠从5.5 天后交媾 (d.p.c.) 到 9.5 d.p.c。使用变频器。测量包括: 数量, 位置和体积的植入点, 以及 inter-implantation 的场地间距;通过心脏活动监测评估胚胎的生存能力。在直接 post-implantation 期间 (5.5 到 8.5 d.p.c.), 妊娠子宫的3维重建在网格和坚实叠加格式使发展的怀孕的视觉表示法在每个子宫垫铁之内成为了。由于基因工程小鼠继续被用来表征从子宫功能障碍中获得的女性生殖表型, 这种方法提供了一种新的方法来检测、量化和表征早期植入事件在体内。这项新的使用3维高频成像显示的能力, 成功地检测, 可视化, 并表征胚胎植入点在早期小鼠怀孕的非侵入性的方式。该技术提供了明显的改善比目前的方法, 这依赖于中断妊娠的总组织和病理特征。在这里, 我们使用视频和文本格式来描述如何成功地执行超声波早期小鼠怀孕, 以产生可靠和重现性的数据与重建的子宫形式的网状和固体3维图像。

如图 1所示, 高频超声可以从5.5 迪普 c 的时间点开始检测植入站点的开发。使用打火机回声 decidualized 子宫内膜作为一个标记的植入点在6.5 迪普 c 允许数量的植入地点和间距这些网站的量化。随着怀孕的进展到 7.5 d.p.c., 一个较暗的回声妊娠囊和胎极也容易辨认。
如图 2所示, 完成后?...

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

三维高频超声对小鼠早期妊娠特征的新应用

老鼠被广泛用于研究妊娠生物学。但是, 这类研究需要终止妊娠, 因为这一问题排除了纵向调查, 必须使用大量动物。因此, 我们描述了一种非侵入性技术的高频超声早期检测和监测的 post-implantation 事件在怀孕的鼠标。

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 ...

Research

JoVE Journal

Developmental Biology

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 ...

科研

发育生物学

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 Mammary Fat Pad Transplantation Technique to Visualize the Vessel Generation of Vascular Endothelial Stem Cells

新型乳腺脂肪移植技术可视化血管内皮干细胞的血管生成

Endothelial cells (ECs) are the fundamental building blocks of the vascular architecture and mediate vascular growth and remodeling to ensure proper ...

Horizontal Whole Mount: A Novel Processing and Imaging Protocol for Thick, Three-dimensional Tissue Cross-sections of Skin

水平整体安装：厚三维组织横截面皮肤的新型加工和成像方案

Processing a tissue of interest to generate a microscopic image that supports a scientific argument can be challenging. The acquisition of high-quality ...

Three-dimensional Rendering and Analysis of Immunolabeled, Clarified Human Placental Villous Vascular Networks

Immunolabeled, 澄清人胎盘长柔毛血管网络的三维渲染与分析

Nutrient and gas exchange between mother and fetus occurs at the interface of the maternal intervillous blood and the vast villous capillary network that ...

Three-dimensional Inflammatory Human Tissue Equivalents of Gingiva

牙龈三维炎症人体组织当量

Periodontal diseases (such as gingivitis and periodontitis) are the leading causes of tooth loss in adults. Inflammation in gingiva is the fundamental ...

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 ...

Three-dimensional Organotypic Cultures of Vestibular and Auditory Sensory Organs

三维脊髓前庭和听觉感官器官的培养

The sensory organs of the inner ear are challenging to study in mammals due to their inaccessibility to experimental manipulation and optical observation. ...

Generation of Scaffold-free, Three-dimensional Insulin Expressing Pancreatoids from Mouse Pancreatic Progenitors In Vitro

Generation of Scaffold-free, Three-dimensional Insulin Expressing Pancreatoids from Mouse Pancreatic Progenitors In Vitro

无支架三维胰岛素表达 Pancreatoids 的实验研究体外培养的小鼠胰祖细胞

The pancreas is a complex organ composed of many different cell types that work together to regulate blood glucose homeostasis and digestion. These cell ...

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 ...