HUMAN TYROSINE HYDROXYLASE AND SCHIZOPHRENIA

这是一份修订后的K02支持竞争续订申请 约翰·W·海考克博士这一奖项将使海科克博士能够继续他的 并继续他最近资助的研究 项目(MH55208)关于精神疾病中的大脑单胺类系统， 它是在本奖项的过程中发起和发展的。 研究与儿茶酚胺功能相关的临床问题的能力 与他之前建立的基础神经科学同时进行 研究计划(NS25134)已经实现了候选人的一个专业 长期目标。因此，候选人的短期目标是 通过招募更年轻的员工专注于实验室的发展 科学家参与实现这两个国家的科学目标 临床和基础研究项目。除了开发 他作为实验室主任的角色，候选人将继续他的 追求新颖的多学科方法实现科学发展 这是他最近开发的，使用不稳定表位的抗体 研究体内信号分子的调控。 拟议的研究计划侧重于酪氨酸羟化酶(TyrOH)和 色氨酸羟基酶(TrpOH)，它催化初始和速度- 限制儿茶酚胺(多巴胺和去甲肾上腺素)的摄入量 5-羟色胺的生物合成。其中的每一个都有变化 系统与精神障碍有牵连，尤其是， 精神分裂症。酪氨酸高度调控--受蛋白质磷酸化 在短期和长期通过转录控制；以及， 选择性剪接(仅在猴子和人类中出现) 产生多种TyrOH亚型，可能还会产生额外水平的 监管。相比之下，人们对TrpOH知之甚少， 尽管它在进化和功能上与TyrOH很接近。尸检 将使用定量印迹分析人脑组织 免疫标记技术和为此开发的一系列抗体库 申请人的目的。酪氨酸和TrpOH蛋白水平，以及 酪氨酸异构体的相对丰度将在 从冷冻条件下解剖出的适合神经化学的脑区 横断面。多巴脱羧酶(紧接在TyrOH和 TrpOH)和多巴胺β-羟基酶(将多巴胺转化为 去甲肾上腺素)蛋白质水平也将被量化，以及类似的 已经为另一类突触前神经元开发了检测方法 单胺能标记物--囊泡单胺和质膜单胺 传送者。主要的学习小组将包括(a.) 确诊为精神分裂症的自杀/猝死受害者 及(B)年龄匹配的猝死对照受试者没有轴1 精神错乱。对主要抑郁症患者的平行、协作研究 将提供比较组，并允许识别潜在的 疾病特定的差异。

This is a revised application for competitive renewal of K02 support for Dr. John W. Haycock. This award will enable Dr. Haycock to continue his scientific development and to pursue his recently funded research program (MH55208) on brain monoaminergic systems in mental illnesses, which was initiated and developed over the course of the current award. The ability to study clinical issues related to catecholamine function concurrently with his previously established fundamental neuroscience research program (NS25134) has realized one of the candidate's major long-term goals. As a result, the candidate's short-term goals are focused upon development of his laboratory by recruiting younger scientists to participate in effecting the scientific goals of both the clinical and fundamental research projects. In addition to developing his role as a laboratory director, the candidate will continue his scientific development by pursuing a novel, multidisciplinary approach which he has recently developed, using antibodies to labile epitopes for studying the regulation of signaling molecules in vivo. The proposed research plan focuses upon tyrosine hydroxylase (TyrOH) and tryptophan hydroxylase (TrpOH), which catalyze the initial and rate- limiting steps in catecholamine (dopamine and norepinephrine) and serotonin biosynthesis, respectively. Alterations in each of these systems have been implicated in mental disorders and, in particular, schizophrenia. TyrOH is highly regulated--by protein phosphorylation in the short-term and by transcriptional control in the long-term; and, alternative splicing (which occurs exclusively in monkeys and humans) produces multiple TyrOH isoforms and perhaps, an additional level of regulation. By contrast, comparatively little is known about TrpOH, despite its evolutionary and functional proximity to TyrOH. Postmortem human brain tissue will be analyzed using quantitative blot immunolabeling techniques and a bank of antibodies developed for this purpose by the applicant. TyrOH and TrpOH protein levels, as well as the relative abundances of TyrOH isoforms, will be measured in neurochemically appropriate brain regions dissected from cryostatic sections. DOPA decarboxylase (immediately downstream of both TyrOH and TrpOH) and dopamine beta-hydroxylase (which converts dopamine to norepinephrine) protein levels will also be quantitated, and similar assays have been developed for another class of presynaptic monoaminergic markers--the vesicular and plasmalemmal monoamine transporters. The primary study groups will consist of (a.) suicide/sudden death victims having confirmed diagnoses of schizophrenia and (b.) age-matched, sudden-death control subjects having no Axis 1 mental disorder. Parallel, collaborative studies of major depressives will provide comparison groups and allow identification of potential disease-specific differences.