The novel matrix structures fractones promote FGF-2 in the neural stem cell niche

新型基质结构 fractone 促进神经干细胞生态位中的 FGF-2

• 批准号: 7387583
• 负责人: FREDERIC MERCIER
• 金额: $ 20.15万
• 依托单位: UNIVERSITY OF HAWAII AT MANOA
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-30 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7387583
• 关键词: Adult; Affect; Affinity; Attention; Basal lamina; Basement membrane; Binding; Blood Vessels; Brain; Bromodeoxyuridine; Cell Proliferation; Cell Surface Receptors; Cells; Confocal Microscopy; Connective Tissue; Count; Coupling; Cues; Data; Degenerative Disorder; Demyelinations; Epidermal Growth Factor; Extracellular Matrix; Extracellular Structure; Fibroblast Growth Factor; Fibroblast Growth Factor 2; Goals; Growth Factor; Growth Factor Interaction; Heparan Sulfate Proteoglycan; Heparin; Heparin Binding; Heparin Binding Growth Factor; Heparitin Sulfate; Immunofluorescence Immunologic; In Vitro; Individual; Injection of therapeutic agent; Inorganic Sulfates; Knockout Mice; Mediating; Meninges; Mitotic; Mus; Nervous System Trauma; Neuroglia; Neurons; Numbers; Organ; Pattern; Peripheral; Play; Procedures; Process; Production; Proliferating; Publishing; Role; Signaling Molecule; Stem cells; Stratum Granulosum; Structure; System; Tissues; Tracer; Transgenic Animals; Transgenic Organisms; Transplantation; Trauma; Unspecified or Sulfate Ion Sulfates; Wild Type Mouse; base; dentate gyrus; design; extracellular; immunocytochemistry; in vivo; lateral ventricle; nerve stem cell; neurogenesis; neuroregulation; novel; perlecan; prevent; receptor; relating to nervous system; research study; response; spatial relationship; stem; sulfation

DESCRIPTION (provided by applicant): Neural stem and progenitor cells (NSPC) offer enormous potential for treating degenerative diseases or traumas of the nervous system. In adult mammalian species, NSC primarily reside in the lateral ventricle walls, where they proliferate and differentiate to produce new neurons and glial cells. It is now evident that microenvironmental cues regulate NSPC proliferation in the "niche" where the cells reside. However, the intrinsic systems that govern the neurogenic niche are poorly understood. Our overall goal is to understand how NSPC, endogenous or transplanted, function within an individual, to eventually replace lost cells. This fundamental understanding is crucial for designing new therapies for neural cells dying due to trauma, degeneration and demyelination. Based on our published data, we hypothesize that the new extracellular structures "fractones" we characterized in direct contact with NSPC in the neurogenic niche of the adult mammalian brain, mediate and regulate the production of new neurons and glial cells throughout adulthood. Our preliminary results indicate that the mechanisms implicated pertain to growth factor/extracellular matrix interactions at the NSPC/fractone interface. The objective of this proposal is to determine whether and how fractones intervene in the process of NSPC proliferation induced by the neurogenic growth factor FGF-2. We found that fractones bind and concentrate FGF-2 in vivo, by a mechanism that is heparan sulfate proteoglycan (HSPG) dependent. Our specific aims are to determine: 1) whether FGF-2 binding to fractones is responsible for FGF-2 induced neurogenesis in the niche; 2) whether NSPC proliferate in contact with fractones that have bound FGF-2; 3) the role of the fractone component perlecan (a major HSPG) on FGF-2-induced NSPC proliferation in the niche. For specific aim 1, we will use biotinylated FGF-2 as a tracer and inducer of NSPC proliferation, and heparatinase, both injected in vivo in the brain of adult mice to determine whether cutting the binding of biotinylated FGF-2 from fractone-borne HSPG annihilates neurogenesis induced by FGF-2. Immunofluorescence will be used to visualize post-mitotic cells that have incorporated bromodeoxyuridine and biotinylated FGF-2. For specific aim 2, we will use biotinylated FGF-2 to identify the spatial relationships between fractones that have bound biotinylated FGF-2 and the emerging cells that have proliferated in response to biotinylated FGF-2. For specific aim 3, we will use transgenic animals (perlecan-null mice) to determine how the lack of perlecan is detrimental to NSPC proliferation induced by biotinylated FGF-2 in the niche. These experiments will allow us to determine whether fractones promote NSPC proliferation induced by FGF-2 in the neural stem cell niche. If demonstrated, fractones will be considered as crucial components that regulate the neural stem cell niche throughout adulthood.

描述(由申请人提供)：神经干细胞和祖细胞(NSPC)为治疗神经系统退行性疾病或创伤提供了巨大的潜力。在成年哺乳动物中，神经干细胞主要存在于侧脑室壁，在那里它们增殖和分化产生新的神经元和神经胶质细胞。现在很明显，微环境信号调节了NSPC在细胞所在的“利基”中的增殖。然而，人们对支配神经源性小生境的内在系统知之甚少。我们的总体目标是了解NSPC，无论是内源性的还是移植的，在个体内如何发挥作用，最终取代丢失的细胞。这一基本认识对于设计治疗因创伤、变性和脱髓鞘而死亡的神经细胞的新疗法至关重要。基于我们已发表的数据，我们假设，在成年哺乳动物大脑的神经源性利基中，我们描述的与NSPC直接接触的新的细胞外结构“分裂”，在成年过程中调节和调节新神经元和神经胶质细胞的产生。我们的初步结果表明，这些机制与生长因子/细胞外基质在NSPC/Fractone界面上的相互作用有关。本研究的目的是确定在神经源性生长因子成纤维细胞生长因子2诱导的NSPC增殖过程中，Fractones是否以及如何进行干预。我们发现，在体内，裂解酮通过一种依赖于硫酸乙酰肝素蛋白多糖(HSPG)的机制结合和浓缩成纤维细胞生长因子-2。我们的具体目的是确定：1)与碎裂酮结合的成纤维细胞生长因子2是否与成纤维细胞生长因子2诱导的神经发生有关；2)神经干细胞是否在与结合了成纤维细胞生长因子2的碎裂酮接触时增殖；3)碎裂酮成分Perlecan(一种主要的热休克蛋白G)在成纤维细胞生长因子-2诱导的神经干细胞增殖中的作用。针对具体目的1，我们将生物素化的成纤维细胞生长因子2作为NSPC增殖的示踪剂和诱导剂，以及肝素酶，在成年小鼠脑内注射，以确定从裂解内酯载体HSPG中切断生物素化的成纤维细胞生长因子-2的结合是否能消除成纤维细胞生长因子-2诱导的神经发生。免疫荧光将被用来观察含有溴脱氧尿苷和生物素标记的成纤维细胞生长因子-2的有丝分裂后细胞。为了达到特定的目的2，我们将使用生物素化的成纤维细胞生长因子-2来确定结合生物素化的成纤维细胞生长因子-2和对生物素化的成纤维细胞生长有反应的新生细胞之间的空间关系。对于特定的目标3，我们将使用转基因动物(Perlecan缺失的小鼠)来确定Perlecan的缺乏如何损害由生物素化的成纤维细胞生长因子-2在利基中诱导的NSPC的增殖。这些实验将使我们能够确定裂解酮是否促进了由成纤维细胞生长因子-2诱导的神经干细胞利基中的NSPC增殖。如果得到证实，分裂音将被认为是在整个成年期调节神经干细胞生态位的关键成分。