Role of the Bradykinin Pathway in Craniofacial Development

缓激肽通路在颅面发育中的作用

• 批准号: 8904316
• 负责人: Laura Anne Jacox
• 金额: $ 4.32万
• 依托单位: HARVARD MEDICAL SCHOOL
• 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 2016-05-16
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8904316
• 关键词: Adult; Anterior; Anterior nares; Antisense Oligonucleotides; Biological Assay; Bradykinin; Bradykinin Receptor; Cell Death; Cell Proliferation; Cells; Cephalic; Complex; Congenital Abnormality; Craniofacial Abnormalities; Data; Defect; Development; Diagnosis; Ectoderm; Elements; Embryo; Endoderm; Etiology; Face; Fertilization; Future; Goals; Hour; Human Biology; Implant; In Situ Hybridization; Jaw; Kininogens; Label; Lead; Ligands; Live Birth; Mammals; Mediator of activation protein; Micromanipulation; Migration Assay; Mission; Modeling; Mothers; National Institute of Dental and Craniofacial Research; Neural Crest; Nitric Oxide; Oral cavity; Pathway interactions; Peptides; Phenotype; Process; Production; Rana; Regulator Genes; Role; Signal Pathway; Signal Transduction; Structure; Testing; Tissues; Tracer; Xenopus; blood pressure regulation; craniofacial; craniofacial development; facial transplantation; in vivo; loss of function; migration; minimally invasive; receptor expression; receptor function

DESCRIPTION (provided by applicant): The broad goal of this proposal is to define signaling pathways that direct craniofacial development, with the long-term goal of diagnosing and treating abnormalities. Specific focus is on the jaws, derived from the neural crest, and on the mouth. The mouth develops from the "extreme anterior domain" (EAD), where the ectoderm and endoderm directly juxtapose. The Bradykinin pathway, previously described only in adults, is locally necessary in the EAD, for mouth formation. However, this pathway also has more global effects on face formation. I hypothesize that the EAD is a craniofacial organizer that regulates mouth and cranial neural crest development, through graded Bradykinin signaling, and culminating in NO production. There are two Aims. Aim 1 will characterize the effects of CPN loss of function on neural crest formation. CPN is a Bradykinin pathway mediator, expressed in the EAD, and loss of function results in abnormal mouth and neural crest tissues. I hypothesize that CPN signaling regulates cranial neural crest determination and serves as a migration stop signal at the facial midline. These hypotheses will be tested after local loss of CPN function using antisense oligonucleotides and a face transplant assay developed in the Sive lab. The effect on neural crest migration will use lineage labeling with fluorescent, injected tracers. Aim will determine the role of Bradykinin signaling on mouth and neural crest formation. CPN processes the ligand Bradykinin to desArgBradykinin. Both are active ligands and lead to nitric oxide NO production. I hypothesize that Bradykinins act in a concentration-dependent manner, through the Bradykinin receptor, to determine the mouth and cranial neural crest. These hypotheses will be tested by implanting beads loaded with Bradykinin peptides into loss of function embryos, and analyzing correction of defects at different distances from the bead. The spectrum of tissues that can respond to Bradykinin signaling will be determined by analysis of Bradykinin receptor function. This study uses the frog Xenopus, an ideal model for analysis of craniofacial defects, as embryos develop outside the mother, and as the developing face is readily accessible. Hundreds of embryos can be obtained, and these allow rapid assays, since mouth and jaw precursors form 36 hours after fertilization, and are functional by 3 days. The large embryos allow micromanipulation, and gain and loss of function assays can be performed. Facial structure and regulatory genes appear conserved between Xenopus and mammals, suggesting that the information gained from this study will be directly relevant to human biology. Craniofacial anomalies are prevalent, appearing in 1 out of 700 live births, yet most have unknown cause. This project is exciting since Bradykinin signaling in facial development has not previously been described, neither has the role of the EAD as a facial organizer. These studies interface with the NIDCR mission, and have potential to provide etiologies for craniofacial defects and suggest future early, minimally invasive treatments.

描述（由申请人提供）：该提案的广泛目标是定义直接颅面发育的信号通路，其长期目标是诊断和治疗异常。特定的重点是源自神经rest和口腔的下颌。嘴从“极端前域”（EAD）发育，外胚层和内胚层直接并置。在EAD中，在EAD中，局部需要仅在成年人中描述的心动激肽途径，以形成嘴。但是，该途径对面部形成也具有更大的全球影响。我假设EAD是一种颅面组织者，可以通过分级的松曲蛋白信号传导来调节口腔和颅神经rest发育，并在无生产中达到顶点。有两个目标。 AIM 1将表征CPN功能丧失对神经rest形成的影响。 CPN是在EAD中表达的心动激肽途径介质，功能丧失会导致口腔和神经rest组织异常。我假设CPN信号传导调节颅神经rest测定，并用作面部中线的迁移停止信号。这些假设将在使用反义寡核苷酸和Sive Lab中开发的面部移植测定法进行局部CPN功能后进行测试。对神经rest迁移的影响将使用谱系标记和荧光，注射的示踪剂。 AIM将确定缓铁蛋白信号在口腔和神经rest形成中的作用。 CPN将布雷蛋白加工为desargbradykinin。两者都是活性配体，并导致一氧化氮无产生。我假设Bradykinins以浓度依赖性的方式通过松曲蛋白受体起作用，以确定口腔和颅神经rest。这些假设将通过植入载有铁丁质肽的珠子来测试这些假设，并分析与珠不同距离的缺陷校正。可以通过缓激肽受体功能的分析来确定可以对心动激肽信号反应的组织光谱。这项研究使用青蛙武士，这是分析颅面缺陷的理想模型，因为胚胎在母亲之外发育，并且随着发育中的面部易于使用。可以获得数百个胚胎，并且可以快速测定，因为受精后36小时形成嘴和下颌前体，并且在3天内起作用。大型胚胎允许微观渗透，并且可以进行功能测定的增益和丧失。面部结构和调节基因似乎在爪蟾和哺乳动物之间保守，这表明从这项研究中获得的信息将与人类生物学直接相关。颅面异常普遍存在，出现在700个活产中的1个，但大多数有未知的原因。该项目令人兴奋，因为Bradykinin在面部发育中的信号传导以前没有被描述过，EAD作为面部组织者也没有作用。这些研究与NIDCR任务接触，并具有为颅面缺陷提供病因的潜力，并暗示未来的早期，微创治疗方法。