ECM and polycystin localization

ECM 和多囊蛋白定位

• 批准号: 8976749
• 负责人: DEANNA M DE VORE
• 金额: $ 2.77万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8976749
• 关键词: Affect; Afferent Neurons; Alleles; Animal Model; Animals; Area; Autosomal Dominant Polycystic Kidney; Caenorhabditis elegans; Cell membrane; Cells; Child; Cilia; Collagen; Data; Defect; Dendrites; Development; Disease; Distal; End stage renal failure; Epithelial; Exhibits; Extracellular Matrix; Extracellular Matrix Proteins; Fibrosis; G Protein-Coupled Receptor Genes; Gene Mutation; Genes; Genetic; Genetic Screening; Goals; Head; Homologous Gene; Human; Ion Channel; Kidney Diseases; Kinesin; Laboratories; Light; Mammalian Cell; Maps; Modeling; Mutate; Mutation; Nematoda; Nervous system structure; Neurons; PKD2 gene; PKD2 protein; Pathway interactions; Pattern; Persons; Phenotype; Play; Property; Protein C; Protein Isoforms; Proteins; Regulation; Research; Role; Sensory Receptors; Site; Sodium Channel; Sorting - Cell Movement; TRPV channel; Tail; Testing; Time; Touch sensation; Zebrafish; base; forward genetics; genome sequencing; improved; insight; knowledge base; male; mating behavior; mutant; polycystic kidney disease 1 protein; public health relevance; receptor; research study; trafficking

DESCRIPTION (provided by applicant): Exploring the role of extracellular matrix components in TRP polycystin ciliary localization and function PKD2 encodes a transient receptor potential polycystin (TRPP) channel receptor protein found in non-motile, primary cilia of mammalian cells. In humans, PKD2 mutations result in Autosomal Dominant Polycystic Kidney Disease (ADPKD). In the nematode Caenorhabditis elegans, the polycystin-2 homolog, PKD-2, localizes to the primary cilia of male-specific sensory neurons (CEMs and RnBs in the head and tail respectively) where it is required for male mating behaviors. Given the ancient and evolutionarily conserved role for polycystin-2 in cilia, C. elegans will be used as a model to identify new genes required for ciliary receptor localization. Mutants were isolated from a forward genetic screen for PKD-2: GFP ciliary localization (Cil) defects. The cil-2(my2) mutants exhibit excess unusual PKD-2 accumulation at the ciliary base, cilium proper, and around the distal dendrite. Three-factor and deficiency mapping, whole genome sequencing, and single gene rescue experiments have determined that the cil-2(my2) allele is a mutation in the gene mec-5. MEC-5 is a unique collagen protein found in the extracellular matrix (ECM) surrounding touch receptor neurons. In touch receptor neurons (TRNs), MEC-1, MEC-5, and MEC-9 are found in the ECM, play a role in mechanosensation, and may localize degenerin/epithelial sodium channels (DEG/ENaCs) along the TRN. Preliminary data indicates that mec-1 and mec-9 regulate PKD-2::GFP localization in male-specific sensory neurons. Therefore current research will determine how these ECM proteins regulate localization and channel properties of the PKD-2 TRP polycystin channel. These studies will shed light on how sensory receptors like PKD-2 are targeted to cilia, and may advance the understanding and treatment of ADPKD.

描述（由申请人提供）：探索细胞外基质成分在 TRP 多囊蛋白纤毛定位和功能中的作用 PKD2 编码在哺乳动物细胞的非运动初级纤毛中发现的瞬时受体电位多囊蛋白 (TRPP) 通道受体蛋白。在人类中，PKD2 突变会导致常染色体显性多囊肾病 (ADPKD)。在线虫秀丽隐杆线虫中，多囊蛋白-2 同源物 PKD-2 定位于雄性特异性感觉神经元（分别位于头部和尾部的 CEM 和 RnB）的初级纤毛，这是雄性交配行为所必需的。鉴于多囊蛋白-2 在纤毛中的古老且进化上保守的作用，线虫将被用作识别纤毛受体定位所需的新基因的模型。通过正向遗传筛选分离出突变体 PKD-2：GFP 纤毛定位 (Cil) 缺陷。 cil-2(my2) 突变体在纤毛基部、纤毛本身和远端树突周围表现出过量的异常 PKD-2 积累。三因素和缺陷图谱、全基因组测序和单基因拯救实验已确定 cil-2(my2) 等位基因是 mec-5 基因的突变。 MEC-5 是一种独特的胶原蛋白，存在于触觉感受器神经元周围的细胞外基质 (ECM) 中。在触觉感受器神经元 (TRN) 中，MEC-1、MEC-5 和 MEC-9 存在于 ECM 中，在机械感觉中发挥作用，并且可能沿着 TRN 定位退化蛋白/上皮钠通道 (DEG/ENaCs)。初步数据表明 mec-1 和 mec-9 调节雄性特异性感觉神经元中的 PKD-2::GFP 定位。因此，当前的研究将确定这些 ECM 蛋白如何调节 PKD-2 TRP 多囊蛋白通道的定位和通道特性。这些研究将揭示 PKD-2 等感觉受体如何靶向纤毛，并可能促进 ADPKD 的理解和治疗。