Name: ultrasonography in experimental reproductive investigations on rats
Uploaded: 2017-12-02T13:00:00.000+00:00
Duration: 7 min 59 s
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。
1. 动物制剂
注: 所有实验均得到穆尔西亚大学动物实验科和穆尔西亚大学妇产科学系的支持。
<ol><li>使用8周老的大鼠 (28 只雌性大鼠) 在所有实验中重200-250 克。 注意: 在这里, 我们还使用了5只怀孕的老鼠。</li>
	<li>为获得孕鼠, 笼八周老雌性和可育雄性大鼠和交配 17:00-23:00 h. 第二天早上阴道堵塞的识别被解释为交配成功.考虑1天的妊娠, 第二天后, 他们交配。</li>
	<li>在怀孕9、15和18天进行实验。</li>
	<li>两组的大鼠, 可以免费获得食物和水, 并保持正常12小时的光照/暗循环。</li>
	<li>在适应到设施条件至少两周后, 使用每天早晨的阴道细胞学来评估发情频率和规律性。 注: 二十八只大鼠与正常4至5天发情阶段选择纳入该研究。</li>
</ol>2. 大鼠成像的制备
注意: 图像是通过高分辨率经腹超声检查得到的。经阴道涂片细胞学检查, 可确定动情周期阶段。
<ol><li>在成像研究之前, 麻醉了2-3% 异氟醚气体在感应室的大坝。</li>
	<li>在超声检查中, 取出动物并立即将鼻口置于与麻醉系统连接的鼻子锥内, 并在 1.5-2% 异氟醚上维持动物。</li>
	<li>将皮草从肋缘移到尾腹, 用快船队和脱毛膏。</li>
	<li>将麻醉大鼠放置在一张加热桌上的仰卧位上, 以确保大鼠的安全, 并在影像疗程期间保证生理参数的最佳舒适和保持。所有的生理参数应与图像和实时采集的数据相结合, 通过超声应用。</li>
	<li>轻轻插入直肠探头 (在润滑后) 以监视体温 (37.5 ° c ±0.5 ° c)。</li>
	<li>将传感器 (30 Hz) 放在固定架上, 并使用手动操纵杆或手将其沿垂直轴和水平轴 (向前-向后和侧面) 移动。</li>
	<li>用石油为基础的眼霜, 以避免干燥过程中的老鼠的眼睛。</li>
</ol>3. 考试程序
注意: rats´生殖器官的解剖: 阴道位于膀胱的背部, 分为两个子宫角, 延伸至肾脏。卵巢通过输卵管连接到子宫角 (图 1)。子宫位于肾脏后部的区域。
<ol><li>
子宫的鉴别
	<ol>
<li>通过用膀胱作为地标来找到宫颈, 跟随宫颈找到左、右子宫角的分支。</li>
		<li>通过选择 "B 模式" 切换到二维查看/视频。用矢状中线图像测量中峡区各子宫角的前后径。使用超声系统软件检测测量。</li>
		<li>在 "B 型" 的矢状中线图像上, 测量子宫内膜腔的子宫内膜厚度, 从回声边界到回声边界。</li>
		<li>通过选择 "彩色多普勒", 改变彩色多普勒模式。使用彩色多普勒识别子宫内膜的血供, 测量子宫内膜的血流量。在彩色多普勒模式下选择以下参数: 高通滤波器设置在4赫兹, 脉冲重复频率设置在4和48赫之间, 脉冲多普勒门设置在0.2 到 0.5 mm 之间。</li>
	</ol>
</li>
	<li>
子宫动脉血流的鉴别
	<ol>
<li>获得多普勒波形的子宫动脉附近的侧下缘的宫内-颈部交界靠近髂动脉两侧。</li>
		<li>在多普勒模式下使用以下参数: 高通滤波器设置在6赫兹, 脉冲重复频率设置在4和48赫之间, 脉冲多普勒门设置在0.2 到 0.5 mm 之间。</li>
		<li>注意对齐血流和多普勒光束以最小化多普勒角。记录多普勒光束和血管之间的夹角。超出60°角度的值是不准确的, 应避免6。</li>
		<li>从三连续周期测量峰值收缩速度和舒张末期速度 (舒张)。然后计算每个子宫角的收缩到舒张期 (舒张) 和阻力指数 (RI) (舒张)/埃因霍温) 值。</li>
		<li>测量5例妊娠大鼠子宫动脉血流量 9th、15th、18th日。</li>
	</ol>
</li>
	<li>
卵巢动脉血流的鉴定 注: 雌性大鼠卵巢位于动物两侧的肾脏侧面, 并居住在子宫角末端的脂肪垫 (图 1)。<ol>
<li>要想象卵巢, 从探针开始在一个横向的平面, 并把它放在动物的侧面, 略低于肋骨。肾脏和脂肪垫有一个回声的外观相比, 卵巢。</li>
		<li>测量卵巢和卵泡的外边界。每个图像的刻度上的数字以毫米为单位, 增量为 0.1 mm。 注: 彩色多普勒模式和功率多普勒模式成像有助于识别卵巢强度和定向流。</li>
	</ol>
</li>
</ol>4. 研究的设计
<ol><li>用阴道涂片细胞学检查发情周期。</li>
	<li>把所有老鼠分成两组。对于组1或前受精 (或 periovulatory), 包括在 proestrus 和发情周期阶段的大鼠。对于2组或后育, 包括早期发情和晚期发情周期阶段的大鼠。</li>
	<li>监测和比较1和2组中峡区各子宫角的前后径。</li>
	<li>监测和比较1和2组子宫内膜的厚度和特征。</li>
	<li>监测和比较的大小 (最大直径) 和特点的卵巢, 并本地化任何 periovulatory 卵泡的两个卵巢组1和2。</li>
	<li>监测和比较1和2组的子宫动脉血流。</li>
	<li>监测和比较怀孕大鼠的子宫动脉血流在不同孕期 (9、15和18的妊娠期)。</li>
	<li>使用 SPSS 执行统计分析. 目前的数据为平均±标准差 (SD) 或中值与分范围。使用不同组之间的学生 t 检验分析结果。小于0.05 的 P 值被认为是统计学上的显著差异。</li>
</ol>

<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>

作者没有什么可透露的。

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><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&amp;nbsp;</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)&amp;nbsp;</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。

大鼠实验性生殖研究中的超声检查

With the development of assisted reproductive technology and the ethical limitations of research on humans, rat animal models have been widely used in ...

ultrasonography-in-experimental-reproductive-investigations-on-rats

Research

JoVE Journal

Medicine

14.2K 次观看.  The First Affiliated Hospital of Anhui Medical University. 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。

视频: Ultrasonography in Experimental Reproductive Investigations on Rats

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

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 routinely used as an in vivo model to study embryo implantation and pregnancy progression. Unfortunately, such murine studies require pregnancy interruption to enable follow-up phenotypic analysis. To address this issue, we used three-dimensional (3-D) reconstruction of HFUS imaging data for early detection and characterization of murine embryo implantation sites and their individual developmental progression in utero. Combining HFUS imaging with 3-D reconstruction and modeling, we were able to accurately quantify embryo implantation site number as well as monitor developmental progression in pregnant C57BL6J/129S mice from 5.5 days post coitus (d.p.c.) through to 9.5 d.p.c. with the use of a transducer. Measurements included: number, location, and volume of implantation sites as well as inter-implantation site spacing; embryo viability was assessed by cardiac activity monitoring. In the immediate post-implantation period (5.5 to 8.5 d.p.c.), 3-D reconstruction of the gravid uterus in both mesh and solid overlay format enabled visual representation of the developing pregnancies within each uterine horn. As genetically engineered mice continue to be used to characterize female reproductive phenotypes derived from uterine dysfunction, this method offers a new approach to detect, quantify, and characterize early implantation events in vivo. This novel use of 3-D HFUS imaging demonstrates the ability to successfully detect, visualize, and characterize embryo-implantation sites during early murine pregnancy in a non-invasive manner. The technology offers a significant improvement over current methods, which rely on the interruption of pregnancies for gross tissue and histopathologic characterization. Here we use a video and text format to describe how to successfully perform ultrasounds of early murine pregnancy to generate reliable and reproducible data with reconstruction of the uterine form in mesh and solid 3-D images.

Mice are widely used to study gestational biology. However, pregnancy termination is required for such studies which precludes longitudinal investigations and necessitates the use of large numbers of animals. Therefore, we describe a non-invasive technique of high-frequency ultrasonography for early detection and monitoring of post-implantation events in the pregnant 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 ...

科研

发育生物学

Fetal Echocardiography and Pulsed-wave Doppler Ultrasound in a Rabbit Model of Intrauterine Growth Restriction

Fetal intrauterine growth restriction (IUGR) results in abnormal cardiac function that is apparent antenatally due to advances in fetoplacental Doppler ultrasound and fetal echocardiography. Increasingly, these imaging modalities are being employed clinically to examine cardiac function and assess wellbeing in utero, thereby guiding timing of birth decisions. Here, we used a rabbit model of IUGR that allows analysis of cardiac function in a clinically relevant way. Using isoflurane induced anesthesia, IUGR is surgically created at gestational age day 25 by performing a laparotomy, exposing the bicornuate uterus and then ligating 40-50% of uteroplacental vessels supplying each gestational sac in a single uterine horn. The other horn in the rabbit bicornuate uterus serves as internal control fetuses. Then, after recovery at gestational age day 30 (full term), the same rabbit undergoes examination of fetal cardiac function. Anesthesia is induced with ketamine and xylazine intramuscularly, then maintained by a continuous intravenous infusion of ketamine and xylazine to minimize iatrogenic effects on fetal cardiac function. A repeat laparotomy is performed to expose each gestational sac and a microultrasound examination (VisualSonics VEVO 2100) of fetal cardiac function is performed. Placental insufficiency is evident by a raised pulsatility index or an absent or reversed end diastolic flow of the umbilical artery Doppler waveform. The ductus venosus and middle cerebral artery Doppler is then examined. Fetal echocardiography is performed by recording B mode, M mode and flow velocity waveforms in lateral and apical views. Offline calculations determine standard M-mode cardiac variables, tricuspid and mitral annular plane systolic excursion, speckle tracking and strain analysis, modified myocardial performance index and vascular flow velocity waveforms of interest. This small animal model of IUGR therefore affords examination of in utero cardiac function that is consistent with current clinical practice and is therefore useful in a translational research setting.

We describe examination of fetal cardiac function with contemporary functional fetal echocardiography and fetoplacental Doppler ultrasound using the VisualSonics VEVO 2100 microultrasound in a surgically induced model of intrauterine fetal growth restriction in a rabbit.

在家兔模型宫内生长受限胎儿超声心动图和多普勒超声脉冲波

Fetal intrauterine growth restriction (IUGR) results in abnormal cardiac function that is apparent antenatally due to advances in fetoplacental Doppler ...

医学

Generation of Topically Transgenic Rats by In utero Electroporation and In vivo Bioluminescence Screening

In utero electroporation (IUE) is a technique which allows genetic modification of cells in the brain for investigating neuronal development. So far, the use of IUE for investigating behavior or neuropathology in the adult brain has been limited by insufficient methods for monitoring of IUE transfection success by non-invasive techniques in postnatal animals. 
 For the present study, E16 rats were used for IUE. After intraventricular injection of the nucleic acids into the embryos, positioning of the tweezer electrodes was critical for targeting either the developing cortex or the hippocampus. 
 Ventricular co-injection and electroporation of a luciferase gene allowed monitoring of the transfected cells postnatally after intraperitoneal luciferin injection in the anesthetized live P7 pup by in vivo bioluminescence, using an IVIS Spectrum device with 3D quantification software. 
 Area definition by bioluminescence could clearly differentiate between cortical and hippocampal electroporations and detect a signal longitudinally over time up to 5 weeks after birth. This imaging technique allowed us to select pups with a sufficient number of transfected cells assumed necessary for triggering biological effects and, subsequently, to perform behavioral investigations at 3 month of age. As an example, this study demonstrates that IUE with the human full length DISC1 gene into the rat cortex led to amphetamine hypersensitivity. Co-transfected GFP could be detected in neurons by post mortem fluorescence microscopy in cryosections indicating gene expression present at &ge;6 months after birth.
 We conclude that postnatal bioluminescence imaging allows evaluating the success of transient transfections with IUE in rats. Investigations on the influence of topical gene manipulations during neurodevelopment on the adult brain and its connectivity are greatly facilitated. For many scientific questions, this technique can supplement or even replace the use of transgenic rats and provide a novel technology for behavioral neuroscience.

Generation of Topically Transgenic Rats by In utero Electroporation and In vivo Bioluminescence Screening

Genes can be manipulated during development of the cortex or the hippocampus of the rat via in utero electroporation (IUE) at E16, to enable fast and targeted modifications in neuronal connectivity for later studies of behavior or neuropathology in adult animals. Postnatal in vivo imaging for control of IUE success is performed by bioluminescence of activating co-transfected luciferase.

外用转基因大鼠通过代<em&gt;在子宫内</em&gt;电穿孔和<em&gt;在体内</em&gt;生物发光筛选

 In utero electroporation (IUE) is a technique which allows genetic modification of cells in the brain for investigating neuronal development. So far, the ...

神经科学

Non-invasive Parenchymal, Vascular and Metabolic High-frequency Ultrasound and Photoacoustic Rat Deep Brain Imaging

Photoacoustics and high frequency ultrasound stands out as powerful tools for neurobiological applications enabling high-resolution imaging on the central nervous system of small animals. However, transdermal and transcranial neuroimaging is frequently affected by low sensitivity, image aberrations and loss of space resolution, requiring scalp or even skull removal before imaging. To overcome this challenge, a new protocol is presented to gain significant insights in brain hemodynamics by photoacoustic and high-frequency ultrasounds imaging with the animal skin and skull intact. The procedure relies on the passage of ultrasound (US) waves and laser directly through the fissures that are naturally present on the animal cranium. By juxtaposing the imaging transducer device exactly in correspondence to these selected areas where the skull has a reduced thickness or is totally absent, one can acquire high quality deep images and explore internal brain regions that are usually difficult to anatomically or functionally describe without an invasive approach. By applying this experimental procedure, significant data can be collected in both sonic and optoacoustic modalities, enabling to image the parenchymal and the vascular anatomy far below the head surface. Deep brain features such as parenchymal convolutions and fissures separating the lobes were clearly visible. Moreover, the configuration of large and small blood vessels was imaged at several millimeters of depth, and precise information were collected about blood fluxes, vascular stream velocities and the hemoglobin chemical state. This repertoire of data could be crucial in several research contests, ranging from brain vascular disease studies to experimental techniques involving the systemic administration of exogenous chemicals or other objects endowed with imaging contrast enhancement properties. In conclusion, thanks to the presented protocol, the US and PA techniques become an attractive noninvasive performance-competitive means for cortical and internal brain imaging, retaining a significant potential in many neurologic fields.

The present work describes a new protocol to perform non-invasive high-frequency ultrasound and photoacoustic based imaging on rat brain, to efficiently visualize deep subcortical regions and their vascular patterns by directing signals on skull foramina naturally present on animal cranium.

非侵入性的脑实质，血管和代谢高频超声和光声大鼠深脑成像

Photoacoustics and high frequency ultrasound stands out as powerful tools for neurobiological applications enabling high-resolution imaging on the central ...

Is My Mouse Pregnant? High-Frequency Ultrasound Assessment

The mouse is the mammalian animal model of choice for many human diseases and biological processes. Developmental biology often requires staged-pregnant mice to determine evolving processes at various timepoints. Moreover, optimal and efficient breeding of model mice requires an assessment of timed pregnancies. Most commonly, mice are mated overnight, and the presence of a vaginal plug is determined; however, the positive predictive value of this technique is suboptimal, and one needs to wait to know if the mouse is truly pregnant. High-resolution ultrasound biomicroscopy is an effective and efficient tool for imaging: 1) Whether a mouse is pregnant; 2) What gestational stage the mouse has reached; and 3) Whether there are intrauterine losses. In addition to the embryos and fetuses, the investigator must also recognize common artifacts in the abdominal cavity so as not to mistake these for a gravid uterus. This article provides a protocol for imaging along with illustrative examples.

High-resolution ultrasound can help streamline experiments requiring timed-pregnant mice by determining the state of pregnancy, gestational age, and pregnancy losses. Presented here is a protocol to illustrate methods to assess mouse pregnancies as well as potential pitfalls (image artifacts) that may mimic pregnancy.

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 ...

Early Detection of Drug-Induced Renal Hemodynamic Dysfunction Using Sonographic Technology in Rats

The kidney normally functions to maintain hemodynamic homeostasis and is a major site of damage caused by drug toxicity. Drug-induced nephrotoxicity is estimated to contribute to 19- 25% of all clinical cases of acute kidney injury (AKI) in critically ill patients. AKI detection has historically relied on metrics such as serum creatinine (sCr) or blood urea nitrogen (BUN) which are demonstrably inadequate in full assessment of nephrotoxicity in the early phase of renal dysfunction. Currently, there is no robust diagnostic method to accurately detect hemodynamic alteration in the early phase of AKI while such alterations might actually precede the rise in serum biomarker levels. Such early detection can help clinicians make an accurate diagnosis and help in in decision making for therapeutic strategy. Rats were treated with Cisplatin to induce AKI. Nephrotoxicity was assessed for six days using high-frequency sonography, sCr measurement and upon histopathology of kidney. Hemodynamic evaluation using 2D and Color-Doppler images were used to serially study nephrotoxicity in rats, using the sonography. Our data showed successful drug-induced kidney injury in adult rats by histological examination. Color-Doppler based sonographic assessment of AKI indicated that resistive-index (RI) and pulsatile-index (PI) were increased in the treatment group; and peak-systolic velocity (mm/s), end-diastolic velocity (mm/s) and velocity-time integral (VTI, mm) were decreased in renal arteries in the same group. Importantly, these hemodynamic changes evaluated by sonography preceded the rise of sCr levels. Sonography-based indices such as RI or PI can thus be useful predictive markers of declining renal function in rodents. From our sonography-based observations in the kidneys of rats that underwent AKI, we showed that these noninvasive hemodynamic measurements may consider as an accurate, sensitive and robust method in detecting early stage kidney dysfunction. This study also underscores the importance of ethical issues associated with animal use in research.

Early stage hemodynamic dysfunction is critical to the development of kidney disease. Yet, detection methodologies are limited. Recent advances in sonography provide a noninvasive, accurate option for early detection of kidney injury. This study outlines a step-by-step, sonographic methodology for detecting kidney dysfunction using a drug-induced nephrotoxicity rat model.

大鼠使用超声技术药物引起的肾血流动力学障碍的早期检测

The kidney normally functions to maintain hemodynamic homeostasis and is a major site of damage caused by drug toxicity. Drug-induced nephrotoxicity is ...

Unraveling the Role of Discrete Areas of the Rat Brain in the Regulation of Ovulation through Reversible Inactivation by Tetrodotoxin Microinjections

Many experimental approaches have been used for studying the role of the brain in the regulation of ovulation. Examples include the lesion and deafferentation of neuronal groups, which are both invasive methods that permanently impair the integrity of the target area. These methods are accompanied by collateral effects that can affect the analysis of acute and temporal regulatory mechanisms. The stereotaxic implantation of guide cannulas aimed at specific brain regions, followed by a recovery period, allows researchers to microinject different drugs after the disappearance of the undesired effects of the surgery. Tetrodotoxin has been used to determine the roles of several brain areas in diverse physiological processes because it transiently inhibits the sodium-dependent action potentials, thus blocking all neural activity in the target region. This protocol combines this method with strategies for the assessment of the estrous cycle and ovulation to reveal the role of discrete brain regions in the regulation of ovulation at particular times of any given stage of the estrous cycle. Awake and unrestrained rats (Rattus norvegicus) were used to avoid the blocking effects that anesthetics and stress hormones exert on ovulation. This protocol can be easily adapted to other species, brain targets and pharmacological agents to study different physiological processes. Future improvements to this method include the design of a microinjection system using glass capillaries of small diameter instead of guide cannulas. This will reduce the amount of tissue damaged during the implantation and decrease the spread of the infused drugs outside the target area.

This protocol describes the construction of a low-cost microinjection system, its stereotaxic implantation into deep-brain structures and the procedure for timed microinjections of tetrodotoxin in awake and unrestrained rats. The goal is to reveal the participation of hypothalamic structures in the regulation of ovulation by inhibiting their neural activity.

通过特洛多毒素微注射可逆灭活，揭开鼠脑离散区在排卵调节中的作用

Many experimental approaches have been used for studying the role of the brain in the regulation of ovulation. Examples include the lesion and ...

Echocardiographic Assessment of Cardiac Anatomy and Function in Adult Rats

The use of experimental animal models has become crucial in cardiovascular science. Most studies using rodent models are focused on two-dimensional imaging to study the cardiac anatomy of the left ventricle and M-mode echo to assess its dimensions. However, this could limit a comprehensive study. Herein, we describe a protocol that allows an assessment of the heart chamber size, left ventricular function (systolic and diastolic) and valvular function. A conventional medical ultrasound machine was used in this protocol and different echo views were obtained through left parasternal, apical and suprasternal windows. In the left parasternal window, the long and short axis were acquired to analyze left chamber dimensions, right ventricle and pulmonary artery dimensions, and mitral, pulmonary and aortic valve function. The apical window allows the measurement of heart chamber dimensions and evaluation of systolic and diastolic parameters. It also allows Doppler assessment with detection and quantification of heart valve disturbances (regurgitation or stenosis). Different segments and walls of the left ventricle are visualized throughout all views. Finally, the ascending aorta, aortic arch, and descending aorta can be imaged through the suprasternal window. A combination of ultrasound imaging, Doppler flow and tissue Doppler assessment have been obtained to study cardiac morphology and function. This represents an important contribution to improve the assessment of cardiac function in adult rats with impact for research using these animal models.

A non-invasive protocol for transthoracic echocardiography assessment of cardiac anatomy and function for adult rats is presented in the current study. The heart valves, all four cardiac chambers and the ascending aorta, aortic arch and descending aorta are studied in detail.

成人大鼠心脏解剖与功能的超声心动图评估

The use of experimental animal models has become crucial in cardiovascular science. Most studies using rodent models are focused on two-dimensional ...

<meta charset="utf8"></head><body>为早期产后疼痛研究创建早产大鼠模型

A Preterm Rat Model for Pain Studies

This research delves into the consequences of consistent pinprick stimulation on preterm offspring to ascertain its long-term implications for pain sensitivity. The primary objective of this protocol was to investigate the impact of neonatal pinprick stimuli on the pain threshold in the later stages of life using a preterm rat model. By establishing this model, we aim to advance the research on understanding and managing early postnatal pain associated with prematurity. The findings of this study indicate that while the baseline thresholds to mechanical stimuli remained unaffected, there was a notable increase in mechanical hypersensitivity following complete Freund's adjuvant (CFA) injection in adult rats. Interestingly, compared with male rats, female rats demonstrated heightened inflammatory hypersensitivity. Notably, maternal behavior, the weight of the litters, and the growth trajectory of the offspring remained unchanged by the stimulation. The manifestation of altered nociceptive responses in adulthood after neonatal painful stimuli could be indicative of changes in sensory processing and the functioning of glucocorticoid receptors. However, further research is needed to understand the underlying mechanisms involved and to develop interventions for the consequences of prematurity and neonatal pain in adults.

<meta charset="utf8"></head><body>用于疼痛研究的早产大鼠模型

Here, we present a concise protocol for creating a preterm rat model, facilitating research on early postnatal pain management. The method involves performing a cesarean section three days before the expected birth, extracting preterm rat pups via hysterectomy, and integrating them with the biological offspring of a surrogate dam.

用于疼痛研究的早产大鼠模型

This research delves into the consequences of consistent pinprick stimulation on preterm offspring to ascertain its long-term implications for pain ...

大鼠实验性生殖研究中的超声检查

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此视频中的章节

三维高频超声对小鼠早期妊娠特征的新应用

在家兔模型宫内生长受限胎儿超声心动图和多普勒超声脉冲波

外用转基因大鼠通过代

非侵入性的脑实质，血管和代谢高频超声和光声大鼠深脑成像

我的老鼠怀孕了吗？高频超声波评估

大鼠使用超声技术药物引起的肾血流动力学障碍的早期检测

通过特洛多毒素微注射可逆灭活，揭开鼠脑离散区在排卵调节中的作用

成人大鼠心脏解剖与功能的超声心动图评估

用于疼痛研究的早产大鼠模型

用于疼痛研究的早产大鼠模型