EXAMINING THE ROLE OF BMAL1, A NOVEL MATERNAL FACTOR IN PREIMPLANTATION DEVELOPMENT

检查 BMAL1（植入前发育中的一种新母体因素）的作用

基本信息

批准号：
10740754
负责人：
Jessica Cassin
金额：
$ 14.89万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-01 至 2025-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10740754
关键词：
ARNTL gene Address Affect Animals Area Bioinformatics Cells Couples Crying Cytoplasm DNA Sequence Alteration Data Defect Deposition Development Early Gene Transcriptions Embryo Embryology Embryonic Development Exhibits Failure Feedback Female Fertilization Fetal Development Foundations Future Genes Genetic Transcription Genome Goals Grant Human Hypothalamic structure Immunofluorescence Immunologic Implant In Situ Hybridization Infertility Investigation Knock-out Live Birth Maternal Messenger RNA Mentors Messenger RNA Methods Modeling Monitor Morphology Movement Mus National Institute of Child Health and Human Development Oocytes Oogenesis Ovary Phase Phenotype Pre-implantation Embryo Development Pregnancy Proteins RNA RNA immunoprecipitation sequencing RNA-Binding Proteins Regulation Regulatory Pathway Reporter Research Role Strategic Planning System Techniques Testing Time Training Translating Translational Regulation Translations Venus Wild Type Mouse Work Writing blastocyst circadian circadian biology circadian regulation embryo culture epitranscriptomics experimental study field study gene network idiopathic infertility insight knock-down live cell imaging mRNA Transcript Degradation molecular clock multiple omics novel offspring oocyte quality preimplantation programs ribosome profiling single-cell RNA sequencing skills transcriptome

项目摘要

PROJECT SUMMARY Unexplained infertility accounts for 25% of infertility cases. One of the challenges in identifying factors related to infertility, particularly unexplained infertility is that there are no live births. A hallmark of genes essential in oogenesis and during preimplantation development is a failure to progress to the point a viable pregnancy can be recognized. As such, identifying genes essential to the these crucial stages is an important step in understating unexplained infertility. Prior research suggests that Bmal1-/- mice produce embryos which are largely lost before the blastocyst stage, even when implanted into pseudopregnant wildtype mice. Dr. Cassin’s preliminary data suggests that initial deficits occur much earlier as Bmal1-/- oocytes are largely observed to be abnormal, likely leading to the phenotype previously described. Further, preliminary work in this grant reveals that BMAL1 is expressed in developing follicles and throughout preimplantation development. BMAL1 has been shown to regulate many of the most essential regulatory functions in early development such as cap-dependent translation, and mRNA regulation, but the role of Bmal1 has not yet been investigated. This grant proposes the examination of Bmal1 during oogenesis and the maternal to zygotic transition (MZT). The overarching hypothesis of this grant is Bmal1 is an essential maternal factor during oogenesis, aids in the regulation of many essential early genes transcribed by the embryo, and regulates a cell-autonomous embryonic clock. Aim1 will address the expression and phenotype of Bmal1 activity during oogenesis. The guiding hypothesis for Aim1 is a primary deficit in oogenesis as a result of Bmal1 m-KO contributes to poor oocyte quality and defects in MZT progression. This Aim will be addressed through the use of embryo culture and immunohistochemical (IHC) techniques. The guiding hypothesis for Aim2 is BMAL1 is an essential factor for preimplantation through the regulation of both maternal mRNAs as well as early embryonic genome activation. As BMAL1 has many regulatory functions Aim2 combine rescue experiments with several -omics techniques to investigate the specific function that BMAL1 has in the MZT. Aim3 examines the regulation of the MZT by Bmal1. The guiding hypothesis of Aim3 is the cell- autonomous clock of the developing embryo is regulated through Bmal1. To address Aim3 live cell imaging of reporter mice will be used to observe movement of BMAL1 throughout the cell. This work specifically focuses on one gene, Bmal1. However, because Bmal1 is a gene involved in many regulatory pathways, this grant opens up a new area of research, the circadian genes in early development. Further, it identifies a novel potential factor in unexplained infertility, and identifies a new direction in the field of infertility. Dr. Cassin will be trained by her mentors in the fields of embryology and circadian biology. Together with training in presenting, scientific writing, and bioinformatics, Dr. Cassin will possess all the skills necessary to establish a successful, independent lab following the mentored phase of the grant.

项目概要不明原因不孕症占不孕症病例的 25%，这是识别相关因素的挑战之一。不孕症，特别是不明原因的不孕症，是没有活产的一个标志。卵子发生和植入前发育期间未能进展到可行妊娠的程度因此，识别这些关键阶段所必需的基因是实现这一目标的重要一步。先前的研究表明 Bmal1-/- 小鼠产生的胚胎大部分是胚胎。即使植入到卡辛博士的假孕野生型小鼠中，在囊胚阶段之前也丢失了。初步数据表明，初始缺陷发生得更早，因为 Bmal1-/- 卵母细胞在很大程度上被观察到此外，这项资助的初步工作揭示了异常，可能导致前面描述的表型。 BMAL1 在发育中的卵泡和整个植入前发育过程中表达。显示可调节早期发育中许多最重要的调节功能，例如上限依赖性翻译和 mRNA 调节，但 Bmal1 的作用尚未得到研究。在卵子发生和母体向合子转变 (MZT) 过程中检查 Bmal1 的总体假设。这笔资助的主要内容是 Bmal1 是卵子发生过程中重要的母体因子，有助于调节许多重要的因素 Aim1 是由胚胎转录的早期基因，并调节细胞自主胚胎时钟。 Aim1 的指导假设是卵子发生过程中 Bmal1 活性的表达和表型。 Bmal1 m-KO 导致卵子发生缺陷，导致卵母细胞质量差和 MZT 进展缺陷。这一目标将通过使用胚胎培养和免疫组织化学 (IHC) 技术来实现。 Aim2 的指导假设是 BMAL1 通过调节两者是植入前的一个重要因素母体 mRNA 以及早期胚胎基因组激活。Aim2 具有许多调节功能。将救援实验与多种组学技术相结合，研究 BMAL1 的具体功能 Aim3 检查 Bmal1 对 MZT 的调节。Aim3 的假设是细胞- 胚胎发育的自主时钟通过 Bmal1 进行调节，以解决 Aim3 活细胞成像问题。报告小鼠将用于观察 BMAL1 在整个细胞中的运动。然而，由于 Bmal1 是一种参与许多调控途径的基因，因此这项资助开放。早期发育中的昼夜节律基因是一个新的研究领域，它还确定了一个新的潜在因素。原因不明的不孕症，并确定了不孕症领域的新方向，卡辛博士将接受她的培训。胚胎学和昼夜节律生物学领域的导师以及演讲、科学写作方面的培训，和生物信息学，卡辛博士将拥有建立成功的独立实验室的所有必要技能在资助的指导阶段之后。