NEURONAL FUNCTIONS OF FHFS

FHFS 的神经元功能

• 批准号: 6993669
• 负责人: MITCHELL GOLDFARB
• 金额: $ 38.2万
• 依托单位: HUNTER COLLEGE
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-08-01 至 2007-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6993669
• 关键词: X ray crystallography; fibroblast growth factor; gene dosage; gene mutation; laboratory mouse; neurogenetics; neuromuscular disorder; sodium channel

DESCRIPTION (provided by applicant): Fibroblast growth factor homologous factors (FHFs) bear sequence and structural homology to fibroblast growth factors (FGFs), but FHFs act solely as neuronal intracellular signalling molecules. FHFs are co-factors for the assembly of the kinase module and are themselves phosphorylated in vivo. FHFs have been shown to associate with a MAP kinase scaffold protein, IB2, and with voltage-gated sodium channels in the central and peripheral nervous systems. Although the precise functions of FHFs are still poorly understood, mice bearing mutations in FHF genes are neurologically impaired. FHF1-/-FHF4 -/- double knockout mice are severely hyperactive and have very poor grip strength. Preliminary electromyography data indicate motor nerve and neuromuscular deficiencies in these animals. Continued research on FHFs shall focus on four new Specific Aims: I) The nerve conduction and neuromuscular transmission defects of FHF1-/-FHF4 -/- mice shall be determined through electrophysiological, ultrastructural, immunological, and biochemical approaches. Detected motor nerve defects shall guide analysis of higher brain centers for similar defects. Genetic interaction between mutant FHF and sodium channel genes shall be investigated. II) The IB2 gene shall be disrupted conditionally in neurons of neonatal mice. The motor phenotypes of IB2 mutant mice shall be determined and compared to those of FHF mutants, and we shall test whether reduced IB2 gene dosage potentiates FHF mutant phenotypes. III) The identity and constituents of brain microvesicles with which phosphorylated FHF associate shall be determined by conventional and affinity-based fractionation methods. IV) The structure of complexes containing FHF together with the FHF-binding segment of IB2 or sodium channels shall be determined by X-ray crystallographic methods. The structures shall guide mutagenesis to determine residues critical for interactions, determine whether IB2 and sodium channels bind FHF by a similar mechanism, and resolve why FHF and FGF folds and surfaces are so paradoxically similar.

描述（由申请人提供）： 成纤维细胞生长因子同源因子（FHF）与成纤维细胞生长因子（FGF）具有序列和结构同源性，但FHF仅作为神经元细胞内信号分子。FHF是用于激酶模块组装的辅因子，并且其自身在体内被磷酸化。FHF已被证明与MAP激酶支架蛋白IB 2以及中枢和外周神经系统中的电压门控钠通道相关。虽然FHF的确切功能仍然知之甚少，但携带FHF基因突变的小鼠神经功能受损。FHF 1-/-FHF 4-/-双敲除小鼠是严重多动的，并且具有非常差的握力。初步的肌电图数据表明，这些动物的运动神经和神经肌肉缺陷。对FHF的继续研究将集中在四个新的具体目标：I）通过电生理学、超微结构、免疫学和生物化学方法确定FHF 1-/-FHF 4-/-小鼠的神经传导和神经肌肉传递缺陷。检测到的运动神经缺陷应指导对类似缺陷的高级脑中心的分析。应研究突变FHF和钠通道基因之间的遗传相互作用。II）IB 2基因应在新生小鼠的神经元中被有条件地破坏。将测定IB 2突变小鼠的运动表型并与FHF突变小鼠的运动表型进行比较，我们将测试降低IB 2基因剂量是否增强FHF突变表型。III）与磷酸化FHF缔合的脑微泡的身份和组成应通过常规和基于亲和力的分级分离方法来确定。IV）含有FHF以及IB 2或钠通道的FHF结合片段的复合物的结构应通过X射线晶体学方法确定。这些结构将指导诱变以确定相互作用的关键残基，确定IB 2和钠通道是否通过类似的机制结合FHF，并解决为什么FHF和FGF折叠和表面如此矛盾地相似。