Functional and evolutionary characterization of the zebrafish ryanodine receptor multigene family

斑马鱼兰尼碱受体多基因家族的功能和进化特征

• 批准号: 312079-2007
• 负责人: Franck, Jens
• 金额: $ 1.82万
• 依托单位: University of Winnipeg
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2011
• 资助国家: 加拿大
• 起止时间: 2011-01-01 至 2012-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=481127
• 关键词: Functional; evolutionary; characterization; zebrafish; ryanodine

The process of gene and whole genome duplication is a fundamental mechanism in the evolution of genome and organism complexity. Genes descending from a duplication event are said to be members of a gene family. The diversity in function of the gene family members is the result of changes in amino acids in the coding region and/or changes in the regulatory domains due to random mutations in the descendant genes. We are studying the genetic regulation and molecular evolution of the intracellular calcium release channels gene family also known as ryanodine receptors (RyR) in non-mammalian vertebrates. We have chosen zebrafish as our model system for several reasons. They are hardy, disease resistant and easily reared and have become a well characterized model for studying vertebrate genetics and development. The zebrafish genome project and recent assembly of sequence data will also permit us to mine for gene sequences. Another distinct advantage of working with fish is the fact that they have discretely compartmentalized muscle fiber types. We have previously shown that fish express fiber type-specific RyR isoforms in skeletal muscle of fish. We are interested in determining the spatial and temporal expression patterns of all members of the RyR gene family in zebrafish. We will clone the regulatory regions of the zebrafish RyR gene isoforms and determine the essential regions for expression using green fluorescent tagged promoter constructs in microinjected zebrafish embryos. We will also investigate the functional role of RyR genes in development by "knocking down" expression by injecting antisense morpholinos into zebrafish embryos. To investigate the molecular evolution of the two-isoform condition in skeletal muscles of fish we will survey the genomic DNA of primitive vertebrate species including coelacanth and hagfish. Comparison of primitive and derived RyR sequences will reveal important information about both the coding and non-coding (regulatory) regions of the genes.

基因和全基因组的复制过程是基因组和生物体复杂性进化的基本机制。来自重复事件的基因被称为基因家族的成员。基因家族成员的功能多样性是由于编码区氨基酸的变化和/或由于后代基因的随机突变而导致的调控结构域的变化。我们正在研究非哺乳动物脊椎动物细胞内钙释放通道基因家族（也称为ryanodine受体（RyR））的遗传调控和分子进化。我们选择斑马鱼作为我们的模型系统有几个原因。它们耐寒、抗病、易饲养，已成为研究脊椎动物遗传和发育的良好模型。斑马鱼基因组计划和最近的序列数据汇编也将使我们能够挖掘基因序列。与鱼一起工作的另一个明显的优势是它们有分散的肌肉纤维类型。我们之前已经证明鱼类在骨骼肌中表达纤维类型特异性的RyR亚型。我们感兴趣的是确定斑马鱼中RyR基因家族所有成员的空间和时间表达模式。我们将克隆斑马鱼RyR基因同工型的调控区域，并在微注射斑马鱼胚胎中使用绿色荧光标记启动子构建物确定表达的必要区域。我们还将通过向斑马鱼胚胎注射反义morpholinos来“敲低”RyR基因的表达，从而研究RyR基因在发育中的功能作用。为了研究鱼类骨骼肌双异构体状态的分子进化，我们将对包括腔棘鱼和盲鱼在内的原始脊椎动物物种的基因组DNA进行研究。比较原始和衍生的RyR序列将揭示基因编码和非编码（调节）区域的重要信息。