Ductin's Role in Craniofacial Patterning and Development

Ductin 在颅面图案形成和发育中的作用

基本信息

批准号：
7786728
负责人：
DANY SPENCER ADAMS
金额：
$ 9.25万
依托单位：
TUFTS UNIVERSITY MEDFORD
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-09-08 至 2010-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7786728
关键词：
Affect Anterior Antibodies Attention Biochemical Biochemistry Biological Biophysics Cells Cellular biology Cephalic Characteristics Chick Embryo Communication Conceptions Congenital Abnormality Connexins Coupled Craniofacial Abnormalities Cytoplasm Data Dental Development Dominant-Negative Mutation Dyes Ear Ectoderm Embryo Embryonic Development Erinaceidae Event Extracellular Space Eye Face Future Gap Junctions Generations Genes Genetic Goals Grant Head Heart Histology Human Image In Situ Hybridization Injection of therapeutic agent Ions Knock-out Knowledge Label Lead Learning Left Life Light Link Localized Mammals Maps Membrane Messenger RNA Microscopy Modeling Molecular Molecular Genetics Monitor Morphogenesis Mus Natural regeneration Nerve Tissue Neural Crest Neurologist Optics Oral Organ Organism Organogenesis Pathway interactions Pattern Phenotype Physiology Play Preventive Principal Investigator Proteins Proton Pump Protons Rana Reagent Regulation Reporting Research Personnel Resolution Role Sample Size Signal Pathway Signal Transduction Skeleton Sonic Hedgehog Pathway Spinal cord damage Staging Structure Study Subject Surveys System Testing Therapeutic procedure Thinking Time Transmembrane Transport United States National Institutes of Health Vertebrates Vesicle Work Xenopus Xenopus frizzled 3 protein Xenopus laevis Zebrafish base bioelectricity comparative concept craniofacial cranium ductin electric field fascinate gain of function genetic manipulation in vivo inhibitor/antagonist insight loss of function member migration novel novel strategies prevent research study scaffold smoothened signaling pathway spatial relationship tumor progression vacuolar H+-ATPase voltage

项目摘要

DESCRIPTION (provided by applicant): Recent experiments provide strong evidence that the protein ductin plays an important role in craniofacial development. Ductin, also known as the 16kD subunit c of V-ATPase, functions in transmembrane transport of protons out of the cytoplasm and into vesicles or the extracellular space; ductin can probably also form gap junctions either alone or with connexins. Thus, ductin has important roles in ion flux across intra-, inter-, and extra-cellular boundaries. We recently discovered that misexpressing wildtype ductin in Xenopus embryos leads to craniofacial abnormalities, as does expressing a dominant negative ductin or injecting antiductin antibodies. The phenotypes of treated embryos, including malformed eyes, ears, and skull, look remarkably like those caused by injection of anti-connexin antibodies and misexpression of the Xenopus frizzled-3 protein. We already know that ductin functions upstream of the Sonic hedgehog signaling pathway; our new results raise the fascinating prospect that ductin and ion flux are also upstream of the Wnt-frizzled cascade during morphogenesis of the vertebrate head. We propose to explore the hypothesis that ductin-based ion flux is an early step in craniofacial morphogenesis that regulates signaling cascades via both V-ATPase dependent proton pumping and cell-cell gap junctional communication. Imaging using in situ hybridization, immuncytochemistry, and in vivo monitoring of pH and fluorescently labeled proteins will be used to characterize ductin's spatial and temporal expression pattern and to monitor the effects of ductin inhibition and misexpression. Loss and gain of function experiments using wildtype and dominant negative ductin constructs will allow us to study the relationship of ductin to downstream genetic pathways. This work will yield important and novel information about the under-studied phenomenon of biophysical control of developmental events, e.g., ion control of secreted signals, thus providing unique and valuable insight into normal morphogenesis and suggesting novel approaches to treating and preventing birth defects.

描述（由申请人提供）：最近的实验提供了有力的证据，表明该蛋白质ductin在颅面发育中起着重要作用。 ductin，也称为V-ATPase的16KD亚基C，在质子从细胞质和囊泡或细胞外空间中的跨膜转运中起作用。 ductin可能也可以单独或与连接蛋白形成间隙连接。因此，ductin在跨离子通量中具有重要作用，跨越细胞内和细胞外边界。我们最近发现，爪蟾胚胎中的野生型ductin会导致颅面异常，表达显性阴性依赖蛋白或注射抗体抗体抗体。所处理的胚胎的表型，包括畸形的眼睛，耳朵和头骨，看起来非常像是由注射抗环性抗体以及对爪蟾毛躁3蛋白的Misexpression引起的表型。我们已经知道，声音刺猬信号通路上游的Ductin功能。我们的新结果提出了一个令人着迷的前景，即脊椎动物头部形态发生期间，ductin和Ion Flux也是Wnt-Frizzed Cascade的上游。我们建议探讨以下假设基于Ductin的离子通量是颅面形态发生的早期一步，可以通过V-ATPase依赖性质子泵送和细胞细胞间隙连接通信来调节信号级联。使用原位杂交，免疫化化学以及对pH值和荧光标记蛋白的体内监测将用于表征ductin的空间和时间表达模式，并监测ductin的抑制和Misexpression的影响。使用野生型和显性阴性ductin构建体的功能实验的丧失和增益将使我们能够研究垂体蛋白与下游遗传途径的关系。这项工作将产生有关生物物理控制事件的生物物理控制现象不足的重要信息，例如，离子对分泌信号的控制，从而为正常的形态发生提供了独特而有价值的见解，并提出了治疗和预防先天缺陷的新方法。