In vivo investigation of fallopian tube transport

项目总结/摘要 本研究的总体目标是研究纤毛在体内输卵管（输卵管）中的作用。到 为此，我的实验室开发了一种高度创新的高分辨率体积光学成像方法 其提供了天然状态下输卵管的前所未有的4D分辨率。 传统上认为纤毛产生卵母细胞/胚胎运输所需的驱动力， 由纤毛病变相关的不育症强化。虽然有研究表明肌肉收缩是 运输的主要因素，最近在纤毛跳动减弱的小鼠模型中进行的研究 证明了排卵卵母细胞的不完全运输，因此，目前围绕 输卵管运输的控制是分开的。然而，很可能纤毛的更细微的整合， 收缩和流动是卵母细胞和胚胎移植所必需的。使用4D体内成像，我们的实验室揭示了一种 输卵管内卵母细胞/胚胎的一些以前未知的活动，表明运输 机制比原先想象的要复杂得多。受精前，上壶腹中的卵母细胞 引导沿着高度纤毛的管腔壁以一致的圆形模式前进， 壶腹由一个“门状”管腔收缩控制。此外，植入前胚胎显示快速，双- 肌肉峡部的定向运动。这表明纤毛的主要作用不太可能是 卵母细胞和胚胎通过输卵管的定向运动。 基于我实验室的初步研究，我认为目前关于纤毛在输卵管中的作用的观点 需要重新评估。该项目的目标是确定纤毛跳动是否是必要的和足够的， 漏斗处的卵母细胞拾取、壶腹中的腔转向以及卵母细胞的定向转移， 胚胎通过输卵管区域。我的实验室拥有独特的功能光学 因此，我们开发了一套光学相干断层扫描仪， (OCT)该方法允许：1）用显微镜对小鼠生殖事件进行活体和动态体积成像， 尺度空间分辨率;（2）纤毛位置和拍频的深度分辨映射;（3） 输卵管收缩波这些测量目前无法用任何其他方法进行， 以前从未与遗传和药物干预结合使用。因此，我有一个 从一个新的和动态的角度探索生殖过程的难得机会。成功完成 这一项目将导致更深入地了解纤毛跳动和肌肉的确切贡献 收缩卵母细胞和胚胎运输在体内，并提供了宝贵的见解难以捉摸的工作原理， 输卵管这项拟议的研究将填补重要的生殖解释和有限的生殖解释之间的差距。 支持这些事件的体内证据。

PROJECT SUMMARY/ABSTRACT The overall objective of this study is to investigate the role of cilia in the fallopian tube (oviduct) in vivo. To accomplish this, my lab has developed a highly innovative, high resolution volumetric optical imaging method that provides unprecendented 4D resolution of the oviduct in its native state. Cilia are traditionally considered to generate the driving force required for oocyte/embryo transport, a theory reinforced by ciliopathy-associated infertility. While there are studies that identify muscular contractions as the predominant factor for transport, recent research conducted in mouse models with weakened ciliary beating demonstrated incomplete transport of ovulated oocytes, and as a result, the current consensus surrounding the control of tubal transport is divided. However, it is likely that a much more nuanced integration of cilia, contractions and flow is required for oocyte and embryo transfer. Using 4D in vivo imaging, our lab revealed a number of previously unknown activities of oocytes/embryos within the oviduct, suggesting that transport mechanisms are much more complex than originally thought. Pre-fertilization, oocytes in the upper ampulla are steered along the highly-ciliated luminal wall in a consistent circular pattern and their progression to the lower ampulla is controlled by a ‘gate-like’ luminal constriction. Furthermore, preimplantation embryos show fast, bi- directional movements in the muscular isthmus. This suggests that it is unlikely that the primary role of cilia is for directional movement of oocytes and embryos through the oviduct. Based on the preliminary studies in my lab, I believe that the current view of the role of cilia in the oviduct requires reassessment. The goal of the project is to determine if ciliary beating is necessary and sufficient for oocyte pick-up at the infundibulum, luminal steering in the ampulla, and the directional transfer of oocytes and embryos through oviductal regions. My lab is enriched with a unique blend of expertise in functional optical imaging and reproductive biology and as a result, we have developed a set of optical coherence tomography (OCT) methods which allow for: 1) live and dynamic volumetric imaging of mouse reproductive events with micro- scale spatial resolution; (2) depth-resolved mapping of cilia location and beat frequency; (3) quantification of the tubal contractile wave. These measurements are currently not possible with any other methods and they have never been applied in conjunction with genetic and pharmacological interventions before. Therefore, I have a rare opportunity to explore reproductive processes from a new and dynamic perspective. Successful completion of this project will lead to a deeper understanding of the exact contributions of ciliary beating and muscular contractions to oocyte and embryo transport in vivo, and provide valuable insights into the elusive workings of the oviduct. The proposed study will fill the gap between important reproductive interpretations and the limited in vivo evidence supporting such events.