HUMAN TYROSINE HYDROXYLASE AND SCHIZOPHRENIA

这份ADAMHA RSDA（II级）的申请书是为John W. Haycock 专注于大脑中的儿茶酚胺系统在心理健康中的作用。 多巴胺能系统与精神分裂症有关， 多巴胺生物合成起始酶和限速酶，酪氨酸 羟化酶（TH），将进行研究。 转录、翻译和 TH的翻译后调节已经在许多 物种 然而，与低等物种不同， TH（mRNA-TH）在人类（和至少一种其他灵长类动物）经历 选择性剪接，导致四种不同的人类mRNA- 日 在申请人的实验室的最近工作中，蛋白质产品 对应于所有这四种mRNA种类的基因 在人肾上腺髓质，而在人和猴黑质纹状体， 系统中，两种形式占主导地位。 因此，在人类和灵长类动物中， TH蛋白不同亚型的改变也可能涉及 调节TH活性。 在本申请中提出的研究涉及分析 多种形式的人类TH。 （A.）区域、细胞和亚细胞 将用免疫印迹法研究不同形式的分布 和使用类型特异性抗体的免疫细胞化学。 区域 不同形式的mRNA-TH和TH蛋白的分布将是 在精神分裂症患者和各种对照组的大脑样本中测定 组（B.）两种主要形式之间的功能差异将是 通过研究位点特异性TH磷酸化的调节来评估 和TH催化活性。Ser 31被Haycock博士鉴定为 调节TH的磷酸化位点。在“正常剪接的”TH中， 申请人最近证明了Ser 31的磷酸化 由MAP 2激酶直接介导，而在另一种形式中， 在脑中，该丝氨酸位于磷酸化共有序列内 钙/钙调蛋白依赖性蛋白激酶II。 对TH的监管将 在人神经母细胞瘤细胞系中原位研究， 表达不同形式的转染细胞系， 用纯化的蛋白激酶和从不同细胞中纯化的TH 线 该奖项将提供的研究开发包括 海考克博士广泛的基础研究 进入人类/临床竞技场。这些研究还将提供 他的专业领域扩展到免疫细胞化学， 分子生物学 在专业成长方面，该奖项将 促进必要的协调努力，使他的多学科 通过防止进一步侵犯部门 和机构职责的时间 的 该奖项还将提高Haycock博士的专业地位， 考虑从副教授晋升为正教授。

This application for an ADAMHA RSDA (Level II) for Dr. John W. Haycock focuses on the role of brain catecholamine systems in mental health. Dopaminergic systems have been implicated in schizophrenia, and the initial and rate-limiting enzyme in dopamine biosynthesis, tyrosine hydroxylase (TH), will be studied. Transcriptional, translational and posttranslational regulation of TH has been demonstrated in a number of species. Unlike in lower species, however, the messenger RNA encoding TH (mRNA-TH) in humans (and at least one other primate) undergoes alternative splicing, resulting in four different species of human mRNA- TH. In recent work from the applicant's laboratory, protein products corresponding to all four of these mRNA species have been demonstrated in human adrenal medulla whereas in the human and monkey nigrostriatal system, two of the forms predominate. Thus, in humans and primates, alterations in the different isoforms of TH protein may also be involved in the regulation of TH activity. The research proposed in this application involves analysis of the multiple forms of human TH. (A.) Regional, cellular and subcellular distributions of the different forms will be studied with immunoblotting and immunocytochemistry using type-specific antibodies. The regional distribution of the different forms of mRNA-TH and TH protein will be determined in brain samples from schizophrenics and various control groups. (B.) Functional differences between the two major forms will be evaluated by studying the regulation of site-specific TH phosphorylation and TH catalytic activity. Ser31 was identified by Dr. Haycock as a regulated phosphorylation site in TH. In "normally spliced" TH, the phosphorylation of Ser31 has recently been demonstrated by the applicant to be directly mediated by MAP2 kinase whereas in the other form abundant in brain, this serine is within a consensus sequence for phosphorylation by calcium/calmodulin-dependent protein kinase II. Regulation of TH will be studied in situ in human neuroblastoma cell lines and in stably transfected cell lines which express the different forms and in vitro with purified protein kinases and TH purified from the different cell lines. The research development that would be provided by this award includes the progression of Dr. Haycock's extensive fundamental research background into the human/clinical arena. The studies will also provide for the expansion of his areas of expertise into immunocytochemistry and molecular biology. In terms of professional growth, the award will facilitate the concerted efforts necessary to bring his multidisciplinary studies to fruition by preventing further encroachment of departmental and institutional responsibilities into Dr. Haycock's research time. The award would also enhance Dr. Haycock's professional status in terms of consideration for promotion from Associate to Full Professor.