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

• 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
• 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

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.

项目摘要 不明原因的不孕症占不孕症病例的25%。在确定与下列因素有关的因素方面的挑战之一是： 不孕症，特别是不明原因的不孕症是没有活产。一个标志性的基因是 卵子发生和植入前发育过程中，未能进展到一个可行的怀孕点， 被认可。因此，确定这些关键阶段所必需的基因是重要的一步， 不明原因的不孕症先前的研究表明，Bmal 1-/-小鼠产生的胚胎主要是 即使植入假孕野生型小鼠，在胚泡阶段之前也会丢失。卡辛医生 初步数据表明，初始缺陷发生得更早，因为Bmal 1-/-卵母细胞在很大程度上被观察到， 异常，可能导致先前描述的表型。此外，这项赠款的初步工作揭示了 BMAL 1在发育中的卵泡和整个着床前发育中表达。BMAL 1已经 在早期发育过程中，它可以调节许多最基本的调节功能，如帽依赖性 翻译和mRNA调控，但Bmal 1的作用尚未研究。这项赠款提出， 在卵子发生和母体到合子转变（MZT）期间检查Bmal 1。总体假设 Bmal 1是卵子发生过程中一个重要的母体因子，有助于调节许多重要的 胚胎转录的早期基因，并调节细胞自主的胚胎时钟。AIM 1将解决 卵子发生过程中Bmal 1活性的表达和表型。Aim 1的指导假设是一个主要的 由于Bmal 1 m-KO导致的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。的 Aim 2的指导假设是BMAL 1是通过调节两个因子来实现植入前的重要因素。 母体mRNA以及早期胚胎基因组激活。由于BMAL 1具有许多调节功能，Aim 2 将联合收割机拯救实验与多组学技术相结合，研究BMAL 1的特异性功能 在MZT中。Aim 3检查Bmal 1对MZT的调节。Aim 3的指导假设是细胞- 发育中的胚胎的自主时钟通过Bmal 1调节。为了解决Aim 3活细胞成像， 报告小鼠将用于观察BMAL 1在整个细胞中的运动。这项工作特别侧重于 一个基因，Bmal 1。然而，由于Bmal 1是一个参与许多调控途径的基因， 开辟了一个新的研究领域，即早期发育中的昼夜节律基因。此外，它确定了一个新的潜在因素， 在不明原因不孕症，并确定了不孕症领域的新方向。卡辛医生将由她训练 胚胎学和昼夜节律生物学领域的导师。再加上演讲，科学写作， 和生物信息学，卡辛博士将拥有建立一个成功、独立实验室所需的所有技能 在赠款的辅导阶段之后。