In vivo investigation of fallopian tube transport

输卵管运输的体内研究

• 批准号: 10537294
• 负责人: Deirdre Scully
• 金额: $ 6.76万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-25 至 2024-01-24
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10537294
• 关键词: Adrenergic beta-Agonists; Affect; Age; Albuterol; Area; Biology; Cilia; Complement; Complex; Consensus; Couples; Dynein ATPase; Embryo; Embryo Transfer; Environment; Epithelial; Event; Female; Fertilization; Frequencies; Genetic; Goals; Image; In Vitro; Infertility; Integral Membrane Protein; Intervention; Investigation; Laboratories; Light; Location; Mammalian Oviducts; Measurement; Methods; Modernization; Molecular Genetics; Movement; Mus; Muscle; Muscle Contraction; Muscle relaxation phase; Oocytes; Optical Coherence Tomography; Pathologic; Pattern; Persons; Pharmacology; Physiological; Physiological Processes; Publications; Regulation; Reproductive Biology; Reproductive Process; Research; Resolution; Role; Site; Smooth Muscle; Technology; Testing; Time; Tube; Tubular formation; United States; Visualization; Woman; base; blastocyst; cell motility; ciliopathy; conditional knockout; constriction; driving force; female reproductive system; fluid flow; image guided; imaging modality; implantation; improved; in vivo; in vivo imaging; innovation; insight; mouse model; optical imaging; optogenetics; pregnant; reproductive; reproductive function; reproductive system disorder; theories; tubal infertility

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.

项目概要/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。 为了实现这一目标，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 它提供了原始状态下输卵管前所未有的 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，这一理论 虽然有研究表明肌肉收缩是导致不孕的原因。 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 排卵卵母细胞的不完全运输，并因此证明了目前围绕 然而，纤毛的整合可能更为微妙。 我们的实验室利用 4D 体内成像揭示了卵母细胞和胚胎移植所需的收缩和流动。 输卵管内卵母细胞/胚胎先前未知的活动数量，表明运输 受精前，上壶腹中的卵母细胞的机制比最初想象的要复杂得多。 沿着高度纤毛的管腔壁以一致的圆形图案引导，并且它们进展到下部 壶腹由“门状”管腔收缩控制。此外，植入前胚胎表现出快速、双收缩。 这表明纤毛的主要作用不太可能是肌肉峡部的定向运动。 卵母细胞和胚胎通过输卵管的定向运动。 根据我实验室的初步研究，我认为目前对纤毛在输卵管中作用的看法 该项目的目标是确定纤毛敲击是否必要且充分。 漏斗部的卵母细胞拾取、壶腹部的管腔转向以及卵母细胞和卵母细胞的定向转移 我的实验室拥有丰富的功能光学专业知识的独特组合。 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) 使用显微技术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）纤毛位置和搏动频率的深度解析映射； 目前，任何其他方法都无法进行这些测量，但它们已经实现了。 以前从未与遗传和药物干预措施一起应用过，因此，我有一个。 从新的动态角度探索生殖过程的难得机会成功完成。 该项目的研究将导致人们更深入地了解纤毛跳动和肌肉运动的确切贡献 卵母细胞收缩和体内胚胎运输，并为难以捉摸的运作提供了宝贵的见解 拟议的研究将填补重要的生殖解释和有限的解释之间的空白。 支持此类事件的体内证据。