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，然后是多个细胞分裂事件以及从母体转录到合子转录的转变。所有这些事件共同作用，建立了第一个因而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是产生滋养外胚层（TE）和内细胞团（ICM），滋养外胚层是胎盘和卵黄囊等胚外结构的前身，而内细胞团则成为胚胎本身。虽然 Oct3/4、Nanog、Sox2、Tead4 和 Cdx2 等转录因子的调控相互作用和形态学贡献已被广泛研究，但令人惊讶的是，人们对分泌生长因子在生长过程中的作用知之甚少。 这些发展的早期阶段。这些早期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发挥功能作用。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 信号传导在植入前阶段胚胎中的重要性。此外，该项目旨在确定 BMP 和谱系规范转录因子之间的分子相互作用，这对于第一次谱系分化至关重要。通过高灵敏度图像分割量化方法，免疫荧光检测和磷酸化 Smad1/5/8 的量化将表明植入前胚胎中活跃的 BMP 信号传导。功能丧失实验将用于确定 BMP 信号传导活性是否会影响在胚芽形成过程中控制谱系规范的两个主要转录网络：Oct4/Nanog/Sox2 的多能调控基序和 Tead4、Cdx2 和 Gata6 的 TE 提交网络。 BMP 信号在植入前哺乳动物胚胎发育中生物学作用的直接证据为理解转录调节因子和信号形成素之间的新型调节相互作用奠定了基础。这种理解有利于多个健康相关学科，如干细胞治疗、癌症生物学和生殖医学。 公共健康相关性：该项目传授 BMP 信号通路的新颖概念知识，这对多个学科都有好处。信号形态发生素和转录因子的相互作用介导这一过程，该过程对于哺乳动物的发育、妊娠以及胚胎和胚胎外干细胞的分化至关重要。增进对这些过程的理解可以通过改进生育治疗以及推进干细胞治疗领域来改善公共健康。