Development and regeneration of the zebrafish maxillary barbel

斑马鱼上颌触须的发育和再生

• 批准号: 8035683
• 负责人: ELIZABETH E LECLAIR
• 金额: $ 41.88万
• 依托单位: DE PAUL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2014-11-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8035683
• 关键词: Abnormal Cell; Adult; Affect; Amputation; Apoptosis; Area; Axon; B-Lymphocytes; Barbus; Behavior; Biological; Biological Assay; Biological Models; Biomedical Research; Blood Vessels; Blood capillaries; Caliber; Candidate Disease Gene; Cell Death; Cell Proliferation; Cell division; Cell physiology; Cells; Cessation of life; Cicatrix; Collaborations; Connective Tissue; Data; Deposition; Development; Electroporation; Embryo; Environment; Epithelial; Esthesia; Extracellular Matrix; Eye; Face; Fertilization; Fishes; Fluid Balance; Funding; Genes; Genetic; Gland; Glypican; Goals; Growth; Health; Heart; Heat-Shock Response; Histocompatibility Testing; Histocytochemistry; Human; Hypertrophic Cicatrix; Image; In Situ Hybridization; Injection of therapeutic agent; Injury; Institution; Investigation; Keloid; Knowledge; Label; Life; Ligands; Longevity; Lymph; Lymphangiogenesis; Lymphatic; Maintenance; Maps; Maxilla; Mediating; Mesenchymal; Methods; Molecular; Molecular and Cellular Biology; Natural regeneration; Nature; Neoplasm Metastasis; Nerve; Nerve Regeneration; Nose; Organ; Pathology; Pathway interactions; Pattern; Peripheral; Peripheral Nerves; Physiological; Pigments; Positioning Attribute; Process; Protocols documentation; Regulation; Research; Resolution; Reverse Transcriptase Polymerase Chain Reaction; Rodent; Role; Sensory; Signal Pathway; Signal Transduction; Signaling Molecule; Skin; Structure; Surveys; System; Tadpoles; Tail; Taste Buds; Techniques; Testing; Tissues; Training; Training Programs; Transgenic Organisms; Trauma; United States National Institutes of Health; Vertebrates; Vibrissae; Wound Healing; Xenopus; Zebrafish; afferent nerve; appendage; base; blastema; capillary; cell behavior; cell type; functional restoration; in vivo; in vivo Model; innovation; light microscopy; melanocyte; migration; mutant; next generation; novel; novel strategies; receptor; repaired; research study; response; response to injury; senescence; skills; tumor

DESCRIPTION (provided by applicant): The zebrafish is a vertebrate animal that can regenerate several adult tissue types. These include well-studied structures such as fins, heart and eyes, but also poorly studied ones, such as the whisker-like facial barbels. The zebrafish maxillary barbel (ZMB) contains skin cells, glands, taste buds, connective tissues, nerves, and vascular components. Because the transparent barbells are external, all of the differentiated cell types within are visible and experimentally accessible. Barbels are known to regenerate in other species of fishes, but have not been studied for this capacity in zebrafish. Based on studies to date, we hypothesize that Wnt signaling molecules and glypican co- receptors are critical for both ZMB outgrowth and regeneration after injury. In our first aim, we will document patterns of cell division and cell death during development and regeneration of this appendage, and confirm the existence and function of the barbel lymphatics, which we are the first to describe. In the second aim, we will 1) map the spatial and temporal resolution of non-canonical Wnt ligands and receptors (glypicans and frizzleds) in ZMB tissue, 2) use transgenic fish to test if restoring glypican function restores ZMB outgrowth in a zebrafish mutant, 3) survey if canonical Wnt signaling is required for ZMB budding and regeneration, and 4) test candidate genes in the ZMB using a morpholino approach. Knowledge gained from these investigations will contribute significantly to our understanding of the genetic interactions involved in the development and maintenance of skin and skin appendages. Because the ZMB is an adult organ that regenerates, this study will also illuminate the response to injury of many terminally differentiated cell types, including blood vessels, lymphatics and nerves. Unlike regeneration in the zebrafish caudal fin, regeneration in the ZMB is characterized by abnormal cell proliferation and extracellular matrix deposition, a process that may be relevant to vertebrate wound healing pathologies such as keloids and/or hypertrophic scars. The ability of zebrafish barbel nerves to regrow axons to their taste bud targets may suggest approaches to restore peripheral sensation to traumatized tissues. Finally, the availability within the ZMB of both a capillary loop and a large, accessible lymphatic is an attractive system in which to study the physiological and pathological functions of the vasculature, including fluid balance and tumor metastasis. ) PUBLIC HEALTH RELEVANCE: This study will establish the biological potential of a novel, easily accessible and relatively non-intrusive system for studying skin appendage development and regeneration in the zebrafish, Danio rerio. Because all vertebrates share many genes in common, findings from this project are expected to relate to the cellular mechanisms of wound healing/scarring, revascularization and nerve regeneration in the adult tissues of other species, such as rodents or humans. The 3-year project will provide undergraduates at our institution with a comprehensive training program in zebrafish genetics, molecular biology and cellular regeneration, all vital skills for the next generation of biomedical professionals.

描述（由申请人提供）：斑马鱼是一种脊椎动物，可以再生几种成年组织类型。这些包括鳍，心脏和眼睛等知识良好的结构，以及研究良好的结构，例如晶须状的面部杠铃。斑马鱼上颌杠铃（ZMB）包含皮肤细胞，腺体，味蕾，结缔组织，神经和血管成分。由于透明的杠铃是外部的，因此内部的所有分化细胞​​类型都是可见的，并且在实验上可以访问。已知杠铃在其他鱼类中再生，但尚未在斑马鱼中研究这种能力。 基于迄今为止的研究，我们假设Wnt信号分子和Glypican共同受体对于损伤后的ZMB产物和再生至关重要。在我们的第一个目标中，我们将记录该附属物发育和再生期间细胞分裂和细胞死亡的模式，并确认杠铃淋巴管的存在和功能，我们是第一个描述的。在第二个目标中，我们将1）在ZMB组织中绘制非传统Wnt配体和受体（Glypicans and Frizzleds）的空间和时间分辨率，2）使用转基因鱼测试是否还原Glypican函数是否恢复ZMB的ZMB在Zebrafish signer中恢复ZMB的测试，并需要ZMB for ZM and tif ZM，3）是否需要ZM，3）IS ZM BUDBEN gyn iS ZM BUDN，如果是ZM BUDN，则需要ZM BUDN，3） ZMB中的候选基因使用吗啡方法。 从这些研究中获得的知识将极大地有助于我们对皮肤和皮肤附属物发展和维持所涉及的遗传相互作用的理解。由于ZMB是一种重生的成年器官，因此这项研究还将阐明许多终末分化细胞类型的损伤的反应，包括血管，淋巴管和神经。与斑马鱼尾鳍的再生不同，ZMB的再生具有异常的细胞增殖和细胞外基质沉积的特征，这一过程可能与脊椎动物伤口愈合的病理学（如乳子状和/或肥大刺伤）有关。斑马鱼杠铃神经能够将轴突重新生成其味蕾靶标的能力可能表明可以恢复外围感觉以恢复受伤的组织。最后，在ZMB内的毛细管环和大型，可及的淋巴处理的可用性是一个有吸引力的系统，在其中研究了脉管系统的生理和病理功能，包括液位平衡和肿瘤转移。 ） 公共卫生相关性：这项研究将确定一种新型，易于访问且相对不受干扰的系统的生物学潜力，以研究斑马鱼Danio Rerio的皮肤附属物发育和再生。由于所有脊椎动物共有许多共同的基因，因此该项目的发现有望与其他物种（例如啮齿动物或人类）成人组织中伤口愈合/疤痕，血运重建和神经再生的细胞机制有关。这项为期3年的项目将在我们机构为本科生提供斑马鱼遗传学，分子生物学和细胞再生方面的全面培训计划，这是下一代生物医学专业人员的所有重要技能。