SULT4A1 Function in Zebrafish Development

描述（由申请人提供）：本提案的总体目标是确定磺基转移酶（SULT）4A 1在发育中的斑马鱼（Danio rerio）胚胎/幼虫模型系统中的调节和功能。SULT 4A 1是一种孤儿酶，基于其结构同源性，已被纳入胞质SULT基因家族。SULT 4A 1蛋白选择性地在所检查的脊椎动物的神经元中表达。SULT 4A 1在神经系统中的功能是令人感兴趣的，因为该蛋白质在哺乳动物中的氨基酸序列大于98.5%相同，并且在脊椎动物物种（包括斑马鱼）中几乎90%相同。在人类中，基于传递不平衡研究及其与精神病理学的关联，SULT 4A 1基因已被鉴定为精神分裂症发生的易感基因。迄今为止，没有令人信服的证据，其底物的选择性或活性已被确定为酶在任何物种或以下细胞或细菌的表达和纯化。斑马鱼SULT 4A 1与其他脊椎动物的SULT 4A 1具有高度同源性，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/幼虫模型允许使用卵注射选择性抑制性吗啉代寡核苷酸来分析SULT 4A 1组织特异性表达、时间调节、酶活性和表达下调的影响。SULT 4A 1 mRNA和蛋白质在4日龄斑马鱼幼虫中表达，表明有可能分析其在胚胎发生和幼虫发育过程中的调控。如以下具体目标所阐述的，对斑马鱼中SULT 4A 1功能和活性的更完整理解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SULT 4A 1基因在斑马鱼胚胎/仔鱼模型系统中的组织特异性表达。目的2研究选择性下调斑马鱼胚胎发育过程中SULT 4A 1表达的形态学和生理学效应。目的3利用斑马鱼胚胎/幼体发育系统研究SULT 4A 1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.