HUMAN TYROSINE HYDROXYLASE AND SCHIZOPHRENIA

这是K 02支持竞争性更新的修订申请， 博士John W.海考克 这一奖项将使海考克博士继续他的 科学发展和追求他最近资助的研究 计划（MH 55208）对脑单胺能系统在精神疾病， 这是在当前的裁决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 研究计划（NS 25134）实现了候选人的主要之一 长期目标。 因此，候选人的短期目标是 他专注于实验室的发展， 科学家参与实现科学目标， 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 通过采用新颖的多学科方法， 这是他最近开发的，使用抗体来对抗不稳定的表位， 研究体内信号分子的调节。 拟议的研究计划侧重于酪氨酸羟化酶（TyrOH）和 色氨酸羟化酶（TrpOH），其催化初始和速率- 儿茶酚胺（多巴胺和去甲肾上腺素）的限制步骤， 血清素生物合成。每一处的变化 系统与精神障碍有关，特别是， 精神分裂症 TyrOH受到蛋白质磷酸化的高度调节 在短期内通过转录控制，在长期内通过转录控制; 选择性剪接（仅发生在猴子和人类中） 产生多种TyrOH同种型，并且可能产生额外水平的 调控 相比之下，对TrpOH的了解相对较少， 尽管其在进化和功能上接近TyrOH。 死后 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抗体库 申请人的目的。 TyrOH和TrpOH蛋白水平以及 TyrOH同种型的相对丰度将在 神经化学上适当的大脑区域从冷冻解剖 路段 多巴脱羧酶（直接位于TyrOH和Dopa脱羧酶的下游） TrpOH）和多巴胺β-羟化酶（其将多巴胺转化为 去甲肾上腺素）蛋白水平也将被定量，并且类似地 已经针对另一类突触前神经元开发了分析方法， 单胺能标记物--囊泡和质膜单胺 运输机 主要研究组将包括（a.） 确诊患有精神分裂症的自杀/猝死者 和（B.）年龄匹配的猝死对照受试者，无轴1 精神障碍 对抑郁症患者的平行合作研究 将提供比较组，并允许识别潜在的 疾病特异性差异。

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.