三维高频超声对小鼠早期妊娠特征的新应用
Summary
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。
Abstract
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。
Introduction
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。
Protocol
Representative Results
Discussion
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…
Disclosures
The authors have nothing to disclose.
Acknowledgements
我们非常感谢蓉赵、洁丽和严营的帮助。
Materials
| VisualSonics Vevo 2100 Ultrasound Imaging Platform/Machine | VisualSonics, inc. | VS-11945 | |
| Vevo Imaging Station | VisualSonics, inc. | SA-11982 | |
| Aquasonic 100 Ultrasound Transmission Gel | Parker | #SKU PLI 01-08 | |
| Isoflurane (IsoThesia) 100mL bottle | Henry Shein | #29404 | |
| PuraLubenAnimal Ophthalmic Ointment | Dechra | #12920060 |
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