In vivo investigation of fallopian tube transport

项目摘要/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。至 为此，我的实验室开发了一种高度创新的、高分辨率的体积光学成像方法 这为处于天然状态的输卵管提供了前所未有的4D分辨率。 纤毛传统上被认为产生卵母细胞/胚胎运输所需的驱动力，这是一种理论 因纤毛疾病相关的不孕不育而强化。虽然有研究将肌肉收缩识别为 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 显示了排卵卵母细胞的不完全运输，因此，目前围绕 对输卵管运输的控制存在分歧。然而，纤毛的更细微的整合很可能是， 卵母细胞和胚胎移植需要收缩和流动。使用4D活体成像，我们的实验室揭示了 输卵管内先前未知的卵母细胞/胚胎活动的数量，表明 机制比最初想象的要复杂得多。受精前，上壶腹的卵母细胞 沿着高度纤毛的管腔壁以一致的圆形图案前进，并向下推进 壶腹是由一种“像门一样”的管腔收缩控制的。此外，植入前的胚胎表现出快速、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为了 卵母细胞和胚胎通过输卵管的定向移动。 根据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充分的 漏斗处的卵母细胞拾取，壶腹的腔转向，以及卵母细胞的定向转移和 胚胎通过输卵管区域。我的实验室拥有丰富的功能光学专业知识 成像和生殖生物学，因此，我们开发了一套光学相干断层扫描 (OCT)方法，允许：1)实时和动态的体积成像小鼠生殖事件与微观 尺度空间分辨率；(2)纤毛位置和拍频的深度分辨映射；(3)纤毛位置和拍频的量化 输卵管收缩波。这些测量目前不可能用任何其他方法，而且他们已经 以前从未与遗传和药物干预一起使用过。因此，我有一个 从新的和充满活力的角度探索生殖过程的难得机会。成功完成 这个项目将使我们更深入地了解睫状肌和肌肉的确切作用。 对体内卵母细胞和胚胎运输的收缩，并提供了对难以捉摸的 输卵管。拟议的研究将填补重要生殖解释与有限生殖解释之间的空白。 支持这种事件的活体证据。

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.