Name: ultrasonography in experimental reproductive investigations on rats
Uploaded: 2017-12-02T13:00:00
Description: Watch this Scientific Journal Video about Ultrasonography in Experimental Reproductive Investigations on Rats at JoVE.com

这项研究得到了穆尔西亚大学动物实验科和穆尔西亚大学妇产科科的支持。我们感谢所有的技术人员在 CEIB (中心实验 en 门多萨市 Biomédicas), 在穆尔西亚大学的动物实验部分, 谁在这个项目上合作。

这项研究是严格按照《关于国家卫生研究院的实验室动物的护理和使用指南》中的建议进行的, 并按照到达 (动物研究: 报告活体实验) 准则进行的。该协议获得了动物实验许可证, 符合指令 2010/63/欧盟的授权编号 A13170404 (Anexo 1)。所有的实验都是在欧洲联盟认证的实验室中进行的, 遵循国家动物伦理准则 (RD 53/2013, 欧盟指令 63/2010)。该议定书得到了穆尔西亚大学动物实验伦理学委员会的批准。
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<ol>
	<li>Hunter, R. K., 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=Adipose-Derived+Stromal+Vascular+Fraction+Cell+Effects+on+a+Rodent+Model+of+Thin+Endometrium.">Adipose-Derived Stromal Vascular Fraction Cell Effects on a Rodent Model of Thin Endometrium.</a> PLoS ONE. 10 (12), e0144823 (2015).</li><li>Wang, H., Dey, S. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Roadmap+to+embryo+implantation:+clues+from+mouse+models.">Roadmap to embryo implantation: clues from mouse models.</a> Nat Rev Genet. 7 (3), 185-199 (2006).</li><li>Pistner, A., Belmonte, S., Coulthard, T., Blaxall, B. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Murine+Echocardiography+and+Ultrasound+Imaging.">Murine Echocardiography and Ultrasound Imaging.</a> J Vis Exp. (42), (2010).</li><li>Lohmiller, J. J., Swing, S. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reproduction+and+Breeding.">Reproduction and Breeding.</a> The Laboratory Rat. , 147-164 (2006).</li><li>Jaiswal, R. S., Singh, J., Adams, G. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-resolution+ultrasound+biomicroscopy+for+monitoring+ovarian+structures+in+mice.">High-resolution ultrasound biomicroscopy for monitoring ovarian structures in mice.</a> Reprod Biol Endocrinol. 7 (69), (2009).</li><li>Kim, G. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Murine+Fetal+Echocardiography.">Murine Fetal Echocardiography.</a> J Vis Exp. (72), (2013).</li><li>Jing, Z., Qiong, Z., Yonggang, W., Yanping, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rat+bone+marrow+mesenchymal+stem+cells+improve+regeneration+of+thin+endometrium+in+rat.">Rat bone marrow mesenchymal stem cells improve regeneration of thin endometrium in rat.</a> Fertil Steril. 101 (2), 587-594 (2014).</li><li>Mu, J., 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=Developmental+changes+in+hemodynamics+of+uterine+artery,+utero-+and+umbilicoplacental,+and+vitelline+circulations+in+mouse+throughout+gestation.">Developmental changes in hemodynamics of uterine artery, utero- and umbilicoplacental, and vitelline circulations in mouse throughout gestation.</a> Am J Physiol Heart Circ Physiol. 291 (3), H1421-H1428 (2006).</li><li>Anderson, C. M., Lopez, F., Zhang, H. Y., Pavlish, K., Benoit, J. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reduced+uteroplacental+perfusion+alters+uterine+arcuate+artery+function+in+the+pregnant+Sprague-Dawley+rat.">Reduced uteroplacental perfusion alters uterine arcuate artery function in the pregnant Sprague-Dawley rat.</a> Biol Reprod. 72 (3), 762-766 (2005).</li><li>Hongmei, 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+Molecular+Imaging+of+Vascular+Endothelial+Growth+Factor+Receptor+2+Expression+for+Endometrial+Receptivity+Evaluation.">Ultrasound Molecular Imaging of Vascular Endothelial Growth Factor Receptor 2 Expression for Endometrial Receptivity Evaluation.</a> Theranostics. 5 (2), 206-217 (2015).</li></ol>

由于这项研究所需的程序性修改和故障排除, 尽管我们的目的是确定的所有阶段的发情周期的大鼠使用超声波, 我们无法找到任何显著的差异。我们推测, 这些困难可能是因为发情周期只持续几天的老鼠, 不同于妇女的周期。我们确信所有的测量都是在正确的时间进行, 以确定任何差异。因此, 我们将发情周期阶段重新组合为两个阶段, 以评估任何差异, 但没有观察到。正如所料, 我们发现每个子宫角?...

大鼠动物模型已广泛应用于生殖医学, 包括胚胎和卵巢移植的1,2。然而, 在过去, 对啮齿动物生殖系统发育的研究是基于对切除组织的一次性组织学检查, 而在大鼠3中, 对日间生殖器官变化的纵向研究是不可能的。超声在人类辅助生殖技术中的应用已有30年的历史, 但这一宝贵的技术在最近才被广泛应用于大鼠。
我们的目标是建立一个评价大大鼠生殖器官的超声方法, 设计生殖和妇科调查的实验模型, 并证明这一程序, 因为据我们所知,目前没有关于此过程的可视化出版物。我们描述了对雌性大鼠生殖系统进行超声检查的程序, 以及用高清晰度超声对解剖和子宫动脉血流的超声表现。我们监测不同发情周期的非妊娠期动物子宫内膜、卵巢和子宫动脉血流的特点, 以评价其在子宫膜厚度、卵巢形态和子宫血流中的显著差异。不同阶段的发情周期, 类似于妇女。我们使用了 70 MHz 频率和分辨率级别为30µm 的高品质超声设备.我们的另一个目的是评估怀孕大鼠子宫血流阻力的变化。这种技术允许研究生殖器官的日常变化而不牺牲动物。
超声对大鼠的应用有若干技术难点。这些困难包括: 大鼠子宫内膜比人类女性更薄4。大鼠卵巢成像困难的原因是大鼠的皮肤和腹壁肌组织较厚, 导致超声5近乎完全衰减, 而子宫动脉在非妊娠期更难发现。鼠.我们已经解决了程序中的许多技术问题, 对于那些仍然存在的问题, 我们将展示如何将它们最小化。
在不需要牺牲动物的情况下, 成功地监测大鼠生殖器官的超声变化, 将为将来建立生殖医学和其他外科手术的动物模型开辟可能性。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Vevo3100 high-resolution in vivo micro-imaging system*</td>
			<td>Visual Sonics, inc.</td>
			<td></td>
			<td>www.visualsonics.com/products</td>
		</tr>
		<tr>
			<td>Vevo integrated rail system including physiological monitoring unit.</td>
			<td>Visual Sonics, inc.</td>
			<td></td>
			<td>www.visualsonics.com/products</td>
		</tr>
		<tr>
			<td>MX400 Transducter</td>
			<td>Visual Sonics, inc.</td>
			<td></td>
			<td>www.visualsonics.com/products</td>
		</tr>
		<tr>
			<td>Vevo Lab Software</td>
			<td>Visual Sonics, inc.</td>
			<td></td>
			<td>www.visualsonics.com/products</td>
		</tr>
		<tr>
			<td>HSD: Sprague Dawley SD</td>
			<td>Envigo, inc.</td>
			<td></td>
			<td>Rat strain</td>
		</tr>
		<tr>
			<td>Lubricating Gel</td>
			<td>General Supply</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>CIBERTEC CA-EAC20 Anesthesia Trolley System</td>
			<td>Cibertec S.A&#160;</td>
			<td></td>
			<td>Anesthesia Machine</td>
		</tr>
		<tr>
			<td>Ecogel 100 ultrasound gel</td>
			<td>Eco-Med Pharmaceuticals Inc.</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Hair removal lotion (Nair)&#160;</td>
			<td>General Supply</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Esteve Veterinaria</td>
			<td></td>
			<td>Inhalatory anesthesia</td>
		</tr>
		<tr>
			<td>* Required software is Vevo software including B-Mode application, pulse wave Doppler application, and vascular strain analysis tools package.</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

ultrasonography in experimental reproductive investigations on rats

随着辅助生殖技术的发展和人类研究的伦理局限性, 大鼠动物模型在生殖医学中得到了广泛的应用。过去, 对啮齿动物生殖系统发育的研究一直是基于对切除组织的一次性组织学检查。近年来, 随着高分辨率经腹超声的发展, 可以对大鼠的生殖器官进行高质量的超声检测, 从而为研究生殖系统提供了一种新的方法。利用高分辨率的超声系统获得图像。对28八周大的非孕鼠和5例妊娠大大鼠进行妇科超声检查。我们描述了在发情周期的不同阶段, 如何在典型的视图中识别生殖系统和相关结构的器官。彩色血流多普勒用于测量子宫动脉血流, 评价不同孕期子宫血流模式的变化。我们已经证明, 超声探测是一种有用的方法来评价变化的内部生殖器官。它的使用提高了进行额外实验的可能性, 包括医疗或外科手术, 并提供了监测超声改变内脏器官而不牺牲动物的能力。

前后子宫角直径或子宫角的两侧子宫内膜厚度 (表 1) 无显著差异。与2组比较, 1 组的平均子宫内膜厚度较厚, 但在两组 (图 3) 之间未发现显著差异 (P > 0.05)。然而, 我们发现子宫内的液体 (在8出28大鼠) 附近的发情周期的变化相关的内膜形态学 (图 2)。
多普勒研究还显示, 非妊娠大鼠子宫角两侧或不同发情周期的血流速度?...

Watch this Scientific Journal Video about Ultrasonography in Experimental Reproductive Investigations on Rats at JoVE.com

Ultrasonography in Experimental Reproductive Investigations on Rats

该手稿描述了超声对雌性大鼠进行生殖和妇科研究的实验模型的效用。一步一步的解释如何进行超声评价显示。

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