SULT4A1 Function in Zebrafish Development

描述（由申请人提供）：本提案的总体目标是建立硫转移酶（SULT） 4A1在发育中的斑马鱼（Danio rerio）胚胎/幼虫模型系统中的调控和功能。SULT4A1是一种孤儿酶，基于其结构同源性已被纳入细胞质SULT基因家族。SULT4A1蛋白在脊椎动物神经元中选择性表达。SULT4A1在神经系统中的功能引起了人们的兴趣，因为该蛋白在哺乳动物中氨基酸序列的相似性超过98.5%，在包括斑马鱼在内的脊椎动物中几乎90%相同。在人类中，SULT4A1基因已被确定为精神分裂症发生的易感基因，基于传播不平衡研究及其与精神病理学的关联。迄今为止，还没有令人信服的证据表明该酶在任何物种中或在细胞或细菌表达和纯化后的底物选择性或活性。斑马鱼SULT4A1与其他脊椎动物中该酶的高度同源性为研究其发育调控和功能提供了有价值的模型系统。在斑马鱼胚胎/幼虫模型中，通过卵注射选择性抑制性morpholino寡核苷酸，可以分析SULT4A1的组织特异性表达、时间调控、酶活性以及下调表达的效果。SULT4A1 mRNA和蛋白在4日龄斑马鱼幼体中表达，显示了分析其在胚胎发生和幼体发育过程中的调控潜力。更全面地了解斑马鱼中SULT4A1的功能和活性，如以下具体目标所阐述的，将为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/幼虫模型系统中的组织特异性发育表达。目的2：探讨选择性下调SULT4A1表达在斑马鱼胚胎发育过程中的形态学和生理作用。目的3利用斑马鱼胚胎/幼虫发育系统研究SULT4A1的活性和底物选择性。

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.