JNK regulation of desmosomes in development.

JNK 对发育过程中桥粒的调节。

• 批准号: 9052712
• 负责人: Amanda Jane Dickinson
• 金额: $ 7.63万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-15 至 2017-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9052712
• 关键词: Adult; Affect; Aftercare; Antibodies; Antisense Oligonucleotides; Antisense Technology; Atomic Force Microscopy; Autoimmune Process; Biological; Biological Assay; Biology; Body Fluids; Bulla; Cell Line; Cell Proliferation; Cell Shape; Chemicals; Co-Immunoprecipitations; Collaborations; Complex; Confocal Microscopy; Craniofacial Abnormalities; Cytoskeletal Proteins; DNA Sequence Alteration; Data; Defect; Desmosomes; Development; Disease; Electron Microscopy; Embryo; Embryonic Development; Epidermis; Epithelial; Family member; Foundations; Future; Gene Expression; Genes; Genetic; Genetic Transcription; Goals; Grant; Health; Heart; Heart Abnormalities; Human; Immune; Immunohistochemistry; Infectious Skin Diseases; Injection of therapeutic agent; JNK-activating protein kinase; JUN gene; Knockout Mice; MAPK8 gene; Maintenance; Mammals; Measures; Mechanical Stress; Mediating; Methods; Mitogen-Activated Protein Kinases; Morphogenesis; Mus; Organ; Phenotype; Phosphorylation; Phosphotransferases; Process; Protein Analysis; Proteins; Proteomics; Rana; Reducing Agents; Regulation; Reverse Transcriptase Polymerase Chain Reaction; Role; Severities; Shapes; Signal Pathway; Signal Transduction; Skin; Skin Abnormalities; Specificity; System; Tadpoles; Techniques; Testing; Tissues; Transplantation; Vertebrates; Work; Wound Healing; Xenopus; Xenopus laevis; craniofacial; desmoplakin; inhibitor/antagonist; innovation; interest; loss of function; novel; photoactivation; protein biomarkers; protein function; research study; response; screening; skin disorder; transcription factor

DESCRIPTION (provided by applicant): During development the epidermis protects the embryo from chemical, biological and mechanical stresses. This organ is also essential in providing tension, necessary for shaping organs while also allowing for cell shape changes necessary for morphogenesis. The integrity of the epidermis is mediated by intercellular junctional complexes, including desmosomes. However, relatively little is known about how desmosomes are regulated during embryonic development. My lab has uncovered a potentially new role for c-Jun NH(2)- terminal kinase (JNK) signaling in regulating desmosomal proteins in the epidermis of Xenopus laevis tadpoles. JNK signaling has been shown to regulate transcription of a variety of genes or to directly phosphorylate cytoskeletal proteins during embryogenesis. But, this MAP kinase family member has never been shown to regulate desmosomal function. We have found that decreased JNK signaling results phenotypes consistent with human conditions resulting from defective desmosomal function such as epidermal fragility and bubbling as well as heart and craniofacial defects. Further, in a proteomics screen for JNK targets we uncovered two plakins; epiplakin and periplakin that are associated with desmosomes in mammals. This proposal aims to use a combination of methods to perform temporal and spatial loss of function of JNK signaling in the epidermis. This will be followed by characterizing the effects in the whole embryo and at the cellular level. We will perform epidermal specific loss of JNK1 function using antisense technology, photoactivation and transplant assays. Desmosomes will be analyzed by electron and confocal microscopy. Finally we aim to determine whether JNK regulates the desmosomal proteins, epiplakin and periplakin, indirectly via transcription or by direct phosphorylation using quantitative PCR and co-immunoprecipitations. I believe this to be an innovative grant that uses whole embryo approaches and bridges developmental signaling and desmosomal function.

描述（由申请人提供）：在发育过程中，表皮保护胚胎免受化学、生物和机械应力。该器官对于提供塑造器官所必需的张力也至关重要，同时还允许形态发生所需的细胞形状变化。表皮的完整性由细胞间连接复合物（包括桥粒）介导。然而，人们对胚胎发育过程中桥粒的调控机制知之甚少。我的实验室发现了 c-Jun NH(2)-末端激酶 (JNK) 信号在调节非洲爪蟾蝌蚪表皮桥粒蛋白方面的潜在新作用。 JNK 信号传导已被证明可以调节多种基因的转录或在胚胎发生过程中直接磷酸化细胞骨架蛋白。但是，这种 MAP 激酶家族成员从未被证明可以调节桥粒功能。我们发现，JNK 信号传导减少导致的表型与人类因桥粒功能缺陷（如表皮脆弱和起泡以及心脏和颅面缺陷）导致的状况一致。此外，在 JNK 靶标的蛋白质组学筛选中，我们发现了两种斑块：与哺乳动物桥粒相关的表斑蛋白和周斑蛋白。该提案旨在使用多种方法的组合来执行表皮中 JNK 信号传导功能的时间和空间丧失。接下来将描述整个胚胎和细胞水平的影响。我们将使用反义技术、光激活和移植检测来进行表皮特异性 JNK1 功能丧失。桥粒将通过电子显微镜和共焦显微镜进行分析。最后，我们的目标是确定 JNK 是否通过转录或通过定量 PCR 和免疫共沉淀直接磷酸化间接调节桥粒蛋白、表皮蛋白和周蛋白。我相信这是一项创新资助，它使用整个胚胎方法并连接发育信号和桥粒功能。

项目成果

Role of JNK during buccopharyngeal membrane perforation, the last step of embryonic mouth formation.

JNK 在胚胎口腔形成的最后一步——颊咽膜穿孔过程中的作用。

• DOI: 10.1002/dvdy.24470
• 发表时间: 2017
• 期刊: Developmental dynamics : an official publication of the American Association of Anatomists
• 作者: Houssin,NathalieS;Bharathan,NavaneethaKrishnan;Turner,StephenD;Dickinson,AmandaJG
• 通讯作者: Dickinson,AmandaJG