Androgen Modulation of Neurodegeneration in Dopamine Neurons

雄激素对多巴胺神经元神经变性的调节

• 批准号: 7986604
• 负责人: REBECCA L CUNNINGHAM
• 金额: $ 2.62万
• 依托单位: UNIVERSITY OF NORTH TEXAS HLTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-03-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7986604
• 关键词: Acute; Address; Age; Androgen Receptor; Androgens; Animal Model; Animals; Apoptosis; Apoptotic; Aromatase; Astrocytes; Attenuated; Award; Behavior; Behavioral; Biological Models; Biology of Aging; Bradykinesia; Cell Line; Cells; Cessation of life; Chemicals; Child; Chronic; Cities; Clinical; Clinical Research; Clinical Trials; Coculture Techniques; Communities; Corpus striatum structure; Data; Doctor of Philosophy; Dopamine; Dopamine Receptor; Dopaminergic Cell; Educational process of instructing; Educational workshop; Embryo; Enrollment; Environment; Enzymes; Estrogens; Face; Fellowship; Foundations; Functional disorder; Gender; Gender Role; Goals; Grant; Health Sciences; High Prevalence; Hormones; Human; Hydrogen Peroxide; In Vitro; Incidence; Individual; Institution; Instruction; Journals; Knockout Mice; Knowledge; LNCaP; Learning; Length; Lesion; Letters; Literature; Manuscripts; Mediating; Methods; Midbrain structure; Minority; Molecular; Monitor; Motor; Muscle Rigidity; Names; Nerve Degeneration; Neuroblastoma; Neurodegenerative Disorders; Neuroglia; Neurons; Operative Surgical Procedures; Oxidative Stress; Oxidopamine; Parkinson Disease; Pathway interactions; Peripheral; Pharmacology; Phase; Play; Population; Preparation; Prevalence; Progress Reports; Progressive Disease; Publications; Rattus; Registries; Reporting; Research; Research Design; Research Personnel; Research Support; Research Training; Resource Sharing; Role; Safety; Scientist; Sex Characteristics; Signal Pathway; Signal Transduction; Site; Stanolone; Substantia nigra structure; Techniques; Testing; Testosterone; Texas; Time; Training; Tyrosine; Tyrosine 3-Monooxygenase; Universities; Vertebrates; Western Blotting; Woman; Work; Writing; aged; behavior measurement; career; caspase-3; design; dopaminergic neuron; embryonic stem cell; experience; field study; human embryonic stem cell; human male; human subject; human subject protection; hydroxyflutamide; immunocytochemistry; improved; in vivo; male; men; neuron loss; neuroprotection; neurotoxicity; nigrostriatal pathway; pre-doctoral; research facility; research study; responsible research conduct; skills; stem; tool

PROBLEM: Parkinson's disease (PD) has a higher prevalence in the human male population. PD is characterized by motor dysfunction, rigidity, and bradykinesia. Current research suggests that gender plays a role in this condition, since males have a greater incidence of PD. It is believed that the underlying mechanism of PD involves oxidative stress leading to cellular apoptosis. In many peripheral cells and some neuronal cells, androgens have been shown to increase apoptosis. PURPOSE: The goal of these studies is to determine androgen's effects on dopaminergic cells following oxidative stress by using both in vitro and in vivo methods. The central hypothesis of this proposal is that androgens increase cellular vulnerability to oxidative stress-induced neurotoxicity in dopaminergic neurons. RESEARCH QUESTIONS: The first specific aim of this proposal is to determine the apoptotic signaling pathways activated by androgens in dopaminergic N27 cells following oxidative stress (hydrogen peroxide). The second specific aim is designed to evaluate the effects of androgens on estrogen-mediated neuroprotection in dopaminergic N27 neurons following oxidative stress. These aims will be accomplished through in vitro molecular studies with and without the presence of astroglia cells and the androgen receptor antagonist, hydroxyflutamide. Lastly, the third specific aim will characterize the in vivo effects of androgens on tyrosine hydroxylaseexpression, neuronal death, and motor behavior in aged male rats exposed to 6-OHDA, which induces oxidative stress and apoptosis in the substantia nigra and striatum. In vivo hormone treatment groups will consist of gonadectomized aged males, gonadally intact aged males, and gonadectomized aged males plus replacement androgen (testosterone or dihydrotestosterone) for either a chronic (3 months) or acute (1 week) treatment time length prior to 6-OHDA lesion. Immunocytochemical and behavioral techniques will be used to accomplish this aim. OUTCOMES: This study will provide basic knowledge on how androgens modulate dopaminergic cellular vulnerability to oxidative stress. Ultimately, this knowledge can be used to provide a foundation to understanding the mechanisms underlying sex differences in neurodegenerative disorders. PHS 416-1 (Rev. 10/05) Page 2 Number pages consecutively at the bottom throughout Form Page 2 the application. Do not use suffixes such as 2a, 2b NAME OF APPLICANT (Last, first, middle initial) Kirschstein-NRSA Individual Fellowship Application Cunningham, Rebecca, L. (To be completed by applicant- follow PHS416-1 instructions) 20. GOALS FOR KIRSCHSTEIN-NRSA FELLOWSHIP TRAINING AND CAREER The training goals of this fellowship include learning scientific approach, troubleshooting, manuscript preparation, and presentation skills. I believe these goals are critical in maturing as a scientist. As a researcher it is of utmost importance to have the ability to not only design and perform an experiment, but to also have the ability to troubleshoot potential problems. The experiments contained within this proposal have been chosen for their ability to test the hypothesis and extend my knowledge in androgen's mechanisms of action by utilizing different experimental techniques and model systems. As a researcher it is imperative to answer "the question." By having a broad knowledge of the tools and techniques available in the scientific community, the reliability of the data obtained is improved. The skills I wish to acquire while undertaking this proposal include Western blot analysis, immunocytochemistry, motor behavior analysis, and stereotaxic surgery. In addition, I wish to expand my knowledge of behavioral studies into the neurodegenerative field of study and determine what impact androgens have on neurodegeneration. 21. ACTIVITIES PLANNED UNDER THIS AWARD: instructions.) Year Research First 80% Second 90% Third 90% Fourth Fifth Approximate percentage of proposed award time in activities identified below. (See Course Work Teaching Clinical 20% 0% 0% 10% 0% 0% 10% 0% 0% PREDOCTORAL FELLOWSHIPS ONLY MD/PhD FELLOWSHIPS ONLY Sixth Briefly explain activities other than research and relate them to the proposed research training. The applicant will participate in several workshops offered by UTHSCSA that include grant and manuscript writing courses. The applicant has been and plans to continue participating in the weekly department seminars and journal clubs to expand her knowledge beyond her field of expertise. During the first year of the fellowship the applicant will take a 1 semester course on Biology of Aging, which is offered by the Department of Pharmacology. 22. TRAINING SITE(S) (organization, city, state) University of Texas Health Science Center-San Antonio, Texas 23. HUMAN EMBRYONIC STEM CELLS Kl No Q Yes If the proposed project involves human embryonic stem cells, list below the registration number of the specific cell line(s) from the following list: http://stemcells.nih.qov/reqistrv/index.asp. Use continuation pages as needed. If a specific line cannot be referenced at this time, include a statement that one from the Registry will be used. Cell Line PHS 416-1 (Rev. 10/05) Page 3 Form Page 3 Kirschstein-NRSA Individual Fellowship Application NAME OF APPLICANT (Last, first, middle initial) Table of Contents Cunningham, Rebecca, L. Page Numbers (Number pages consecutively at the _ _ bottom throughout the application. Section 1 - Applicant Do not use suffixes such as6a, 6b.) Face Page 1 Sponsor's Contact Information, Description (Form Page 2) 2 Training & Career Goals, Activities Planned Under This Award, Training Site, Human Embryonic Stem Cells (Form Page 3) 3 Table of Contents (Form Page 4) 4 Biographical Sketch - Applicant/Fellow (Not toexceed four pages) 5-6 Previous Research Experience (Form Page 5) 7-9 Research Training Plan Introduction to Revised Application (not to exceed 1page) A. Specific Aims TV. ^ 10 B. Background/Significance !(......(Not to exceed 10pages) J 11 C. Preliminary Studies/Progress Report ../T | 12 D. Research Design andMethods ).. ^ 13 E. Human Subjects (Required if Item 9 on the Face Page is marked "Yes") 20 Protection of Human Subjects (Required if Item 9 on the Face Page is marked "Yes") - Data and Safety Monitoring Plan (Required if Item 9 on the Face Page is marked "Yes" and a Phase I, II, or III clinical trial is proposed - Inclusion of Women and Minorities (Required if Item 9 on the Face Page is marked "Yes" and is Clinical Research) - Targeted/Planned Enrollment Table (for new and continuing clinical research studies) - Inclusion of Children (Required if Item 9 on the Face Page is marked "Yes") - F. Vertebrate Animals (Required if Item 10on the Face Page is marked "Yes") 20 G. Literature Cited 20 H. Resource Sharing 24 I. Respective Contributions 24 J. Selection of Sponsor and Institution 24 K. Responsible Conduct of Research 24 Section 2 - Sponsor's/Co-Sponsor's Information Biographical Sketch-Sponsor 25 Research Support Available 29 Previous Trainees 31 Training Plan, Environment, Research Facilities 31 Number of Fellows/Trainees to be Supervised 32 Applicant's Qualifications and Potential 32 Checklist (Completed by Fellow/Applicant & Sponsoring Institution) 33 Section 3 - References (Minimum of 3) (See instructions for submission of references.) List full name, institution, and department of individuals submitting reference letters. 1) Marilyn Y. McGinnis, Ph.D., UTHSCSA, Pharmacology 2) William P. Clarke, Ph.D., UTHSCSA, Pharmacology 3) J. Randy Strong, Ph.D., UTHSCSA, Pharmacology Other Items (list): Personal Data Page for Fellowship Applicants Section 4 - Appendix (5 collated sets. Nopage numbering necessary. Not to exceed 3 publications; 2 for predoctoral candidates.) E3 Check if Appendix is included PHS 416-1 (Rev. 10/05) Page 4 Form Page 4

问题：帕金森病 (PD) 在男性人群中的患病率较高。 PD 是 其特征为运动功能障碍、强直和运动迟缓。目前的研究表明性别发挥作用 在这种情况下，男性的 PD 发病率更高。据信，底层 PD的机制涉及氧化应激导致细胞凋亡。在许多外周细胞和一些 神经元细胞，雄激素已被证明可以增加细胞凋亡。目的：这些研究的目标是 通过体外和体内实验确定雄激素对氧化应激后的多巴胺能细胞的影响 体内方法。该提案的中心假设是雄激素增加了细胞对 氧化应激诱导的多巴胺能神经元的神经毒性。研究问题：第一个具体问题 该提案的目的是确定雄激素激活的细胞凋亡信号通路 氧化应激（过氧化氢）后的多巴胺能 N27 细胞。第二个具体目标的设计 评估雄激素对雌激素介导的多巴胺能 N27 神经元神经保护的影响 氧化应激后。这些目标将通过体外分子研究来实现 不存在星形胶质细胞和雄激素受体拮抗剂羟基氟他胺。最后， 第三个具体目标将表征雄激素对酪氨酸羟化酶表达的体内影响， 暴露于 6-OHDA（诱导氧化应激）的老年雄性大鼠的神经元死亡和运动行为 以及黑质和纹状体的细胞凋亡。体内激素治疗组将包括 性腺切除的老年男性、性腺完整的老年男性和性腺切除的老年男性 替代雄激素（睾酮或二氢睾酮）用于慢性（3 个月）或急性（1 周）6-OHDA 损伤前的治疗时间长度。免疫细胞化学和行为技术将 用于实现此目的。结果：这项研究将提供有关雄激素如何作用的基本知识 调节多巴胺能细胞对氧化应激的脆弱性。最终，这些知识可以用来 为理解神经退行性疾病中性别差异的机制提供基础 失调。 PHS 416-1（修订版 10/05）第 2 页 在整个表格第 2 页的底部连续编号页码 该应用程序。请勿使用 2a、2b 等后缀 申请人姓名（姓氏、名字、中间名首字母） Kirschstein-NRSA 个人奖学金申请 坎宁安，丽贝卡，L. （由申请人填写 - 请遵循 PHS416-1 说明） 20. Kirschstein-NRSA 奖学金培训和职业目标 该奖学金的培训目标包括学习科学方法、故障排除、手稿 准备和演讲技巧。我相信这些目标对于成为一名成熟的科学家至关重要。作为一个 对于研究人员来说，最重要的是不仅要有能力设计和执行实验，还要有能力 也有能力解决潜在的问题。该提案中包含的实验 被选中是因为他们有能力检验这一假设并扩展我对雄激素机制的了解 利用不同的实验技术和模型系统采取行动。作为一名研究者，必须 回答“问题”。通过对科学领域可用的工具和技术有广泛的了解 社区，提高了所获得数据的可靠性。我希望在从事这项工作时获得的技能 建议包括蛋白质印迹分析、免疫细胞化学、运动行为分析和立体定位 外科手术。此外，我希望将我的行为研究知识扩展到神经退行性疾病领域 研究并确定雄激素对神经变性的影响。 21. 本奖项计划开展的活动： 指示。） 年研究 前 80% 第二个90% 第三 90% 第四 第五 拟议奖励时间在下列活动中的大致百分比。 （看 临床课程教学 20% 0% 0% 10% 0% 0% 10% 0% 0% 仅限博士前奖学金 仅限医学博士/博士奖学金 第六 简要解释研究以外的活动，并将其与拟议的研究培训联系起来。 申请人将参加 UTHSCSA 提供的多个研讨会，其中包括资助和手稿 写作课程。申请人已经并计划继续参加每周部门 研讨会和期刊俱乐部将她的知识扩展到她的专业领域之外。第一年期间 奖学金申请者将参加为期 1 个学期的衰老生物学课程，该课程由 药理学系。 22. 培训地点（组织、城市、州） 德克萨斯大学健康科学中心 - 德克萨斯州圣安东尼奥 23. 人类胚胎干细胞 Kl 否 Q 是 如果拟议项目涉及人类胚胎干细胞，请在下面列出以下列表中特定细胞系的注册号： http://stemcells.nih.qov/reqistrv/index.asp。根据需要使用延续页面。 如果此时无法引用特定行，请包含一项声明，表明将使用注册表中的某一行。 细胞系 PHS 416-1（修订版 10/05）第 3 页 表格第 3 页 Kirschstein-NRSA 个人奖学金申请 申请人姓名（姓氏、名字、中间名首字母） 目录 丽贝卡·L·坎宁安 (Cunningham) 页码 （在页码处连续编号 _ _ 贯穿整个应用程序的底部。 第 1 部分 - 申请人请勿使用 6a、6b 等后缀。） 正面第 1 页 赞助商的联系信息、说明（表格第 2 页）2 培训和职业目标、该奖项计划的活动、培训地点、人力 胚胎干细胞（表格第3页）3 目录（表格第 4 页）4 简历 - 申请人/研究员（不超过四页）5-6 以往的研究经历（表格第5页）7-9 研究培训计划 修改后的申请简介（不超过1页） A. 具体目标电视。 ^10 B. 背景/意义！(......(不超过10页) J 11 C. 初步研究/进展报告../T | 12 D. 研究设计和方法).. ^ 13 E. 人类受试者（如果主页上的第 9 项标记为“是”则为必填项）20 保护人类受试者（如果主页上的第 9 项标记为“是”则为必需）- 数据和安全监控计划（如果首页第 9 项标记为“是”则需要 并提议进行 I、II 或 III 期临床试验 - 纳入妇女和少数民族（如果主页上的第 9 项标记为“是”则为必填项） 是临床研究）- 目标/计划入组表（用于新的和持续的临床研究）- 包含儿童（如果主页上的第 9 项标记为“是”，则为必填项）- F. 脊椎动物（如果首页第 10 项标记为“是”则为必填项） 20 G. 被引文献 20 H. 资源共享 24 一、各自的贡献 24 J. 赞助商和机构的选择 24 K. 负责任的研究行为 24 第 2 部分 - 赞助商/联合赞助商的信息 传记素描赞助商 25 提供研究支持 29 历届学员 31 培训计划、环境、研究设施 31 受指导的研究员/实习生人数 32 申请人的资格和潜力 32 清单（由研究员/申请人和赞助机构填写）33 第 3 部分 - 参考文献（至少 3 篇） （参见提交参考文献的说明。） 列出提交推荐信的个人的全名、机构和部门。 1) Marilyn Y. McGinnis，博士，UTHSCSA，药理学 2) William P. Clarke，博士，UTHSCSA， 药理学 3) J. Randy Strong，博士，UTHSCSA，药理学 其他项目（清单）： 奖学金申请人的个人数据页面 第 4 节 - 附录 （5 套整理集。无需页码编号。出版物不得超过 3 份；博士前候选人不得超过 2 份。） E3 检查是否包含附录 PHS 416-1（修订版 10/05）第 4 页 表格第 4 页