In vivo analysis of mammalian fertilization

哺乳动物受精的体内分析

基本信息

批准号：
10311522
负责人：
Irina Larina
金额：
$ 60.48万
依托单位：
BAYLOR COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-01-19 至 2023-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10311522
关键词：
3-Dimensional Animal Model Animals Area Assisted Reproductive Technology Behavior Biological Cells Cilia Clinical Complex Contraceptive methods Contrast Media Data Defect Development Embryo Embryo Transfer Ensure Environment Estradiol Estrous Cycle Event Failure Female Female infertility Fertility Fertilization Fertilization failure Flagella Frequencies Genetic Models Histologic Hormonal Hormones Human Image Imaging Device Individual Infertility Invertebrates Investigation Knockout Mice Laboratories Lead Light Location Longitudinal Studies Male Infertility Mammalian Oviducts Maps Measurement Mediating Methods Modeling Molecular Molecular Genetics Movement Mus Muscle Contraction Oocytes Optical Coherence Tomography Organ Ovulation Pathology Pattern Peer Review Phase Positioning Attribute Pregnancy Procedures Process Progesterone Protocols documentation Publications Publishing Recording of previous events Reproduction Reproductive Biology Reproductive Process Research Research Design Resolution Scanning Sea Urchins Speed Sperm Motility Sperm Tail Superovulation System Time Visualization Zona Pellucida base blastocyst cell motility early pregnancy egg experimental study high resolution imaging hormone regulation human model imaging genetics imaging modality imaging study imaging system in vivo in vivo imaging infertility treatment innovation insight intraperitoneal mouse model optical imaging preimplantation reproductive reproductive tract research study sex sperm cell

项目摘要

PROJECT SUMMARY/ABSTRACT Estrous cycle, ovulation, fertilization, and pre-implantation pregnancy are fundamental reproductive processes of clinical importance. While research has shed light on the cellular and molecular mechanisms mediating these events, much of these data are derived from static histological analysis, low-resolution visualizations, and studies of invertebrate models (e.g. sea urchin). Therefore, any conclusions regarding mammalian fertilization, which takes place deep inside the body, are extrapolated and do not necessarily represent the native state. If this technical limitation was overcome, we may gain a more complete understanding of mammalian reproduction leading to the development of better fertility treatments and Assisted Reproductive Technologies (ART). By integrating expertise in live, functional optical coherence tomography (OCT) and reproductive biology, we recently established a set of unique methods for in vivo imaging of the female mouse reproductive tract. Our approach allows for (i) live, dynamic volumetric imaging of the mouse Fallopian tube (oviduct) with micro- scale spatial resolution, (ii) depth-resolved mapping of oviduct cilia location and cilia beat frequency (CBF); and (iii) tracking of individual sperm and their motility within the oviduct. None of these measurements are currently possible with other methods, and the dynamic environment of the female reproductive tract is too complex to model. Therefore, we are in a unique position to directly visualize specific mammalian reproductive processes from an entirely new vantage point. We propose the first in vivo volumetric imaging study of mammalian fertilization. This study is taking advantage of new technological developments in OCT imaging and will allow for quantitative assessment of hormonal regulation of oviduct cilia beating and muscle contractions, and functional analysis of fertility failures in mouse models of human defects. This study will likely provide new insight on the process of mammalian fertilization in its native state and lead to a better understanding of pathologies resulting in infertility. It will also establish new functional live imaging tools, which will be a major step forward in reproductive research. Scientific Premise, Scientific Rigor, and Relevant Biological Variables: This proposal is aimed to fill a significant gap in the field of reproductive biology through highly innovative live imaging methods, which we developed. All proposed experiments are supported by strong preliminary data, which have been published in four peer-reviewed publications; one more publication is currently under review. We carefully articulated the number of experimental animals to be used, and the rationale for the choice of the models. “Sex as a biological variable” does not apply to our study design. Extensive details and references to our published protocols are provided to ensure that preliminary and proposed experiments can be replicated in other laboratories.

项目摘要/摘要发情周期，排卵，受精和植入前妊娠是基本复制过程临床重要性。虽然研究已经阐明了介导的细胞和分子机制这些事件，其中大部分数据来自静态组织学分析，低分辨率可视化，和无脊椎动物模型（例如海胆）的研究。因此，关于哺乳动物的任何结论施肥发生在体内的深处，被外推，不一定代表本地国家。如果克服了这种技术限制，我们可能会对哺乳动物的繁殖导致发展更好的生育治疗和辅助生殖技术（艺术）。通过将专业知识整合到现场功能性光学相干断层扫描（OCT）和生殖生物学中，我们最近，建立了一组独特的方法，用于对雌性小鼠复制道的体内成像。我们的方法允许（i）使用微型 - 小鼠输卵管（Oviduct）实时的动态体积成像刻度空间分辨率，（ii）纤毛纤毛位置和纤毛频率（CBF）的深度分辨映射；和（iii）跟踪单个精子及其在输卵管中的运动。这些测量当前都不是使用其他方法可能，女性复制道的动态环境太复杂了模型。因此，我们处于独特的位置，可以直接可视化特定的哺乳动物重复过程从一个全新的有利位置。我们提出了哺乳动物施肥的第一个体内体积成像研究。这项研究正在利用 OCT成像中的新技术发展，将允许对Horsemonal进行定量评估纤毛纤毛殴打和肌肉收缩的调节，以及小鼠生育失败的功能分析人类缺陷的模型。这项研究可能会提供有关哺乳动物受精过程的新见解它的原生状态并导致对病理的更好理解，导致不育症。它也将建立新的功能性实时成像工具，这将是复制研究的重要一步。科学前提，科学严谨和相关的生物学变量：该建议旨在填补通过高度创新的实时成像方法，生殖生物学领域的显着差距，我们发达。所有提出的实验均由强大的初步数据支持，这些数据已发表在四个经同行评审的出版物；目前正在审查另一份出版物。我们仔细阐明了使用的实验动物数量以及选择模型的基本原理。 “作为生物学的性可变”不适用于我们的研究设计。广泛的详细信息和对我们发布的协议的参考是提供的目的是确保可以在其他实验室中复制初步和建议的实验。