SULT4A1 Function in Zebrafish Development

描述(申请人提供)：本提案的总体目标是在斑马鱼(Danio Rerio)胚胎/仔鱼发育模型系统中建立磺基转移酶(Sult)4A1的调节和功能。SULT4A1是一种孤儿酶，根据其结构同源性已被归入胞质sult基因家族。SULT4A1蛋白在所研究的脊椎动物的神经元中选择性表达。SULT4A1在神经系统中的功能令人感兴趣，因为该蛋白在哺乳动物中的氨基酸序列同源性超过98.5%，在包括斑马鱼在内的脊椎动物中的同源性几乎为90%。在人类中，SULT4A1基因已被确定为精神分裂症发生的易感基因，这是基于传递不平衡研究及其与精神病理学的关联。到目前为止，还没有令人信服的证据表明TH酶在任何物种中或随后的细胞或细菌表达和纯化中具有底物选择性或活性。斑马鱼SULT4A1与其他脊椎动物的SULT4A1酶具有很高的同源性，为研究其发育调控和功能提供了一个有价值的模型系统。斑马鱼胚胎/仔鱼模型可以分析SULT4A1的组织特异性表达、时间调节、酶活性以及通过鸡蛋注射选择性抑制吗啉寡核苷酸下调表达的效果。SULT4A1基因在4天龄斑马鱼幼体中表达，为分析其在胚胎发生和幼体发育过程中的调控奠定了基础。更全面地理解SULT4A1在斑马鱼中的功能和活性，如以下具体目的所阐述的，将为了解其在人类神经生物学中的作用提供有价值的见解。目的1分析SULT4A1在斑马鱼胚胎/仔鱼模型系统中的组织特异性发育表达。目的研究选择性下调SULT4A1基因在斑马鱼胚胎发育过程中的形态和生理效应。目的3利用斑马鱼胚胎/仔鱼发育系统研究SULT4A1的活性和底物选择性。 与公共卫生相关：磺基转移酶(Sult)4A1是一种神经元表达的酶，在包括斑马鱼在内的脊椎动物中高度保守。这种酶在大脑中有选择性地表达，但没有已知的底物或功能，尽管它与人类精神分裂症的发展有基因上的联系。将通过抑制SULT4A1在胚胎发育过程中的表达来研究SULT4A1在遗传上可驯化的斑马鱼胚胎/幼体发育系统中的调节和功能。

DESCRIPTION (provided by applicant): The overall goal of this proposal is to establish the regulation and function of sulfotransferase (SULT) 4A1 in the developing zebrafish (Danio rerio) embryo/larval model system. SULT4A1 is an orphan enzyme that has been included in the cytosolic SULT gene family based upon its structural homology. SULT4A1 protein is selectively expressed in neurons in the vertebrates examined. The function of SULT4A1 in the nervous system is of interest since the protein is greater than 98.5% identical in amino acid sequence in mammals and almost 90% identical in vertebrate species, including zebrafish. In humans, the SULT4A1 gene has been identified as a susceptibility gene for the occurrence of schizophrenia based on both transmission disequilibrium studies and its association with psychopathology. To date, no convincing evidence as to its substrate selectivity or activity has been identified for th enzyme in any species or following cell or bacterial expression and purification. The high homology of zebrafish SULT4A1 to the enzyme in other vertebrates provides a valuable model system for the examination of its developmental regulation and function. The zebrafish embryo/larval model allows for the analysis of SULT4A1 tissue specific expression, temporal regulation, enzymatic activity and effects of down-regulation of expression using egg injection of selective inhibitory morpholino oligonucleotides. SULT4A1 mRNA and protein are expressed in 4-day old zebrafish larvae demonstrating the potential to analyze its regulation during embryogenesis and larval development. A more complete comprehension of SULT4A1 function and activity in zebrafish, as elaborated by the following specific aims, will provide valuable insights into its role in human neurobiology. Aim 1 To analyze the tissue-specific developmental expression of SULT4A1 in the zebrafish embryo/larval model system. Aim 2 To investigate the morphological and physiological effects of selective down-regulation of SULT4A1 expression during zebrafish embryonic development. Aim 3 To investigate the activity and substrate selectivity of SULT4A1 using the zebrafish embryo/larval developmental system. PUBLIC HEALTH RELEVANCE: Sulfotransferase (SULT) 4A1 is a neuronally expressed enzyme that is highly conserved in vertebrates including zebrafish. The enzyme is selectively expressed in the brain but has no known substrates or function although it has been associated genetically with the development of schizophrenia in humans. The regulation and function of SULT4A1 will be investigated in the genetically tractable zebrafish embryo/larval developmental system using inhibition of its expression during embryonic development.