Detection of BMP Signaling In Pre-Implantation Mouse Embryos

植入前小鼠胚胎中 BMP 信号传导的检测

• 批准号: 8400805
• 负责人: Nabora Soledad Reyes de Barboza
• 金额: $ 3.58万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8400805
• 关键词: Address; BMP4; Binding; Biological; Biological Assay; Bone Morphogenetic Proteins; CDX2 gene; Cancer Biology; Cell Adhesion; Cell Communication; Cell Polarity; Cells; Chemicals; Commit; Complication; Data; Data Set; Defect; Detection; Development; Discipline; Embryo; Embryonic Development; Event; Extraembryonic Structure; Failure; Fertility; Fertilization; Foundations; Genetic Transcription; Growth Factor; Health; Hour; Image; Immunofluorescence Immunologic; In Vitro; Inner Cell Mass; Knowledge; Light; Maintenance; Mediating; Methods; Molecular; Morphogenesis; Morula; Mus; Nucleic Acid Regulatory Sequences; Pattern; Placenta; Pregnancy; Pregnancy loss; Process; Public Health; Reproductive Medicine; Resolution; Role; Signal Pathway; Signal Transduction; Staging; Stem cells; System; Testing; Therapeutic; Transcript; Work; Yolk Sac; base; blastocyst; bone morphogenetic protein receptor type II; bone morphogenetic protein receptors; cell fate specification; chromatin immunoprecipitation; embryo culture; embryo stage 2; embryonic stem cell; feeding; imaging Segmentation; improved; inhibitor/antagonist; loss of function; member; morphogens; novel; pluripotency; preimplantation; research study; spatiotemporal; transcription factor

DESCRIPTION (provided by applicant): Mammalian development begins with fertilization, followed by multiple cell cleavage events and the transition from maternal to zygotic transcription. All these events work in concert to establish the first and thus highly significant developmental event, lineage differentiation. The first step in the establishment of different lineages in the pre-implantation embryo gives rise to the trophectoderm (TE), the precursor for extraembryonic structures such as the placenta and yolk sac, and the inner cell mass (ICM), which becomes the embryo proper. While the regulatory interactions and morphological contributions of transcription factors such as Oct3/4, Nanog, Sox2, Tead4, and Cdx2 have been extensively studied, surprisingly little is known about the roles of secreted growth factors during these early stages of development. The presence of bone morphogenetic proteins (BMPs) in these early mouse embryonic stages suggests likely functional roles for BMP signaling in lineage differentiation. This project will determine the significance of BMP signaling in pre- implantation stage embryos by elucidating the spatiotemporal emergence and functional role of BMP signaling. Also, this projects aims to identify molecular interactions between BMPs and lineage specification transcription factors vital for the first lineage differentiation. Through a highly sensitive image segmentation quantification method, immunofluorescence detection and quantification of phosphorylated Smad1/5/8 will denote active BMP signaling in pre-implantation embryos. Loss of function experiments will be used to determine if BMP signaling activity impinges on two major transcriptional networks that control lineage specification during blastulation: the pluripotent regulatory motifs of Oct4/Nanog/Sox2 and the TE committing networks of Tead4, Cdx2, and Gata6. Direct evidence for the biological role of BMP signaling in pre-implantation mammalian embryonic development provides a foundation for understanding novel regulatory interactions between transcriptional regulators and signaling morphogens. This understanding benefits multiple health related disciplines such as stem cell therapeutics, cancer biology, and reproductive medicine. PUBLIC HEALTH RELEVANCE: This project imparts novel conceptual knowledge of BMP signaling pathways, which is beneficial to multiple disciplines. Interactions of signaling morphogens and transcription factors mediate this process that are vital in mammalian development, gestation, and differentiation of embryonic and extraembryonic stem cells. Advancing the understanding of these processes can improve public health by allowing for improved fertility treatments as well as advancing the field of stem cell therapeutics.

描述（由申请人提供）：哺乳动物的发育始于受精，随后是多个细胞裂解事件以及从母体到二元转录的过渡。所有这些事件都可以协同建立第一个且因此非常重要的发展事件，即谱系分化。在植入前胚胎中建立不同谱系的第一步会引起滋养剂（TE），这是胚胎外结构（例如胎盘和蛋黄囊）的前体，以及内部细胞质量（ICM），这成为胚胎适当的。尽管经过广泛研究了转录因子，例如Oct3/4，Nanog，Sox2，Tead4和CDX2等转录因子的调节相互作用和形态学贡献，但令人惊讶的是，对于分泌生长因子的作用知之甚少。 这些发展的早期阶段。在这些早期的小鼠胚胎阶段中骨形态发生蛋白（BMP）的存在表明BMP信号在谱系分化中可能作用。该项目将通过阐明BMP信号传导的时空出现和功能作用来确定BMP信号传导在植入前胚胎中的重要性。此外，该项目旨在确定BMP和谱系规范转录因子之间对第一个谱系分化至关重要的转录因子之间的分子相互作用。通过高度敏感的图像分割定量方法，磷酸化SMAD1/5/8的免疫荧光检测和定量将表示植入前植入胚胎中的主动BMP信号。功能实验的丧失将用于确定在爆破过程中控制谱系规范的两个主要转录网络上的BMP信号传导活动是否会影响：OCT4/NANOG/SOX2的多能调节基序以及TEAD4，CDX2，CDX2和GATA6的TE构成网络。直接证据证明了BMP信号传导在植入前哺乳动物胚胎发育中的生物学作用为理解转录调节剂和信号传导形态的新调节相互作用提供了基础。这种理解使多个与健康相关的学科受益，例如干细胞疗法，癌症生物学和生殖医学。 公共卫生相关性：该项目赋予了BMP信号通路的新颖概念知识，这对多个学科有益。信号传导形态和转录因子的相互作用介导了该过程，这在哺乳动物发育，妊娠和胚胎和胚外干细胞的分化中至关重要。促进对这些过程的理解可以通过改善生育治疗以及推进干细胞疗法领域来改善公共卫生。