REPRODUCTIVE STEROIDS AND EXPERIMENTAL STROKE

生殖类固醇和实验性中风

• 批准号: 6718432
• 负责人: Stephanie J Murphy
• 金额: $ 10.03万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-05-03 至 2005-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6718432
• 关键词: brain injury; cerebral ischemia /hypoxia; dopamine; dopamine receptor; dopamine transporter; estrogens; gender difference; hormone regulation /control mechanism; hormone therapy; immunocytochemistry; in situ hybridization; infarct; laboratory rat; microdialysis; neural transmission; neuroprotectants; neurotransmitter metabolism; northern blottings; progesterone; reproductive hormone; steroid hormone; stroke; western blottings

Candidate: Dr. Stephanie J. Murphy plans to establish a biomedical research career integrating neuroscience and specialization in comparative medicine with the clinical aspects of stroke. Her extensive formal doctoral training in biochemistry, clinical training in comparative medicine, and postdoctoral research in cerebral ischemia has given her a broad multidisciplinary background to achieve this goal. Environment: The A/CCM Research Division at the Johns Hopkins Medical Institutions offers exceptional personal guidance, interactive and experienced researchers, and excellent physical facilities with which to execute this research proposal. Drs. Patricia D. Hurn and Richard J. Traystman have offered their support and mentorship to Dr. Murphy. Research: We have shown that brain outcomes after experimental stroke are gender-specific and linked to reproductive hormone status. There are striking differences in post-ischemic brain injury after middle cerebral artery occlusion (MCAO) in female vs. male rat, suggesting that females enjoy substantial neuroprotection when confronted with an ischemic episode. Preliminary work suggests that reproductive steroids may reduce brain injury in older animals. The work proposed will clarify the neuroprotective importance of estrogen, alone or with progesterone, in reproductively senescent female (RSF) ischemic rat brain and assess if estrogen or combined hormone therapy alters ischemic outcome by reducing injury due to dopaminergic hyperactivity and hydroxyl radical formation. In Aim 1, we will determine if hormone replacement provides neuroprotection in RSF rats during reversible MCAO. Ischemic outcomes are determined by infarction volume analysis. Aim 2 determines if estrogen or combined hormone treatment attenuates ischemic injury by reducing dopaminergic neurotransmission utilizing microdialysis. Aim 3 determines if reproductive steroids alter infarction outcomes after MCAO in RSF rats by reducing hydroxyl radical formation from either dopamine (DA)-induced or non-DA derived sources as determined by salicylate trapping and microdialysis. In Aim 4, we will assess if ovarian steroids induce DA-modulated neuroprotection through increases in DA uptake and intracellular transport via increased expression of the DA membrane transporter (DAT) and the vesicular monoamine transporter (VMAT2), respectively. DAT and VMAT2 expression are determined using in situ hybridization, Northern blots, RNAase protection assays, immunohistochemistry, and Western blots.

候选人：斯蒂芬妮·J·墨菲（Stephanie J. Murphy）博士计划建立一项生物医学研究职业，该研究将比较医学中的神经科学和专业化与中风的临床方面相结合。 她在生物化学，比较医学方面的临床培训以及脑缺血的博士后研究中进行了广泛的正式博士培训，这给了她广泛的多学科背景，以实现这一目标。环境：约翰·霍普金斯医学机构的A/CCM研究部提供了出色的个人指导，互动和经验丰富的研究人员，并提供了出色的物理设施，可以执行这项研究建议。 博士。 Patricia D. Hurn和Richard J. Traystman已向墨菲博士提供了支持和指导。研究：我们已经表明，实验中风后的大脑结局是性别特异性的，并且与生殖激素状态有关。 雌性与雄性大鼠中脑动脉阻塞（MCAO）后缺血后脑损伤存在明显的差异，这表明雌性在面对缺血性发作时享有实质性神经保护作用。初步工作表明，生殖类固醇可以减少老年动物的脑损伤。 提出的工作将阐明雌激素单独或孕激素在生殖性衰老女性（RSF）缺血大鼠脑中的神经保护重要性，并评估雌激素或组合激素治疗是否通过减少多巴胺疗法过度和羟基自由基形成而导致的缺血性损伤改变了缺血性损伤。 在AIM 1中，我们将确定在可逆MCAO期间，激素替代量是否在RSF大鼠中提供神经保护作用。 缺血性结果由梗塞体积分析确定。 AIM 2通过减少使用微透析的多巴胺能神经传递来确定雌激素或激素治疗联合治疗缺血性损伤。 AIM 3通过减少来自多巴胺（DA）诱导的或非DA衍生的源源的RSF大鼠MCAO后的生殖类固醇是否改变了MCAO后MCAO的梗塞结果，这是通过水杨酸酯捕获和微二聚体确定的。 在AIM 4中，我们将评估卵巢类固醇是否通过增加DA膜转运蛋白（DAT）和囊泡单胺转运蛋白（VMAT2）的表达来增加DA摄取和细胞内转运的神经保护作用。 使用原位杂交，北印迹，RNAase保护测定，免疫组织化学和蛋白质印迹确定DAT和VMAT2表达。

项目成果

The human gonadotropin releasing hormone type I receptor is a functional intracellular GPCR expressed on the nuclear membrane.

• DOI: 10.1371/journal.pone.0011489
• 发表时间: 2010-07-08
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Re M;Pampillo M;Savard M;Dubuc C;McArdle CA;Millar RP;Conn PM;Gobeil F Jr;Bhattacharya M;Babwah AV
• 通讯作者: Babwah AV

Trafficking and quality control of the gonadotropin releasing hormone receptor in health and disease.

• DOI: 10.1016/j.mce.2008.10.051
• 发表时间: 2009-02-27
• 期刊: Molecular and cellular endocrinology
• 影响因子: 4.1
• 作者: Conn PM;Janovick JA
• 通讯作者: Janovick JA

X-chromosome inactivation in monkey embryos and pluripotent stem cells.

• DOI: 10.1016/j.ydbio.2012.08.009
• 发表时间: 2012-11-15
• 期刊: Developmental biology
• 影响因子: 2.7
• 作者: Tachibana M;Ma H;Sparman ML;Lee HS;Ramsey CM;Woodward JS;Sritanaudomchai H;Masterson KR;Wolff EE;Jia Y;Mitalipov SM
• 通讯作者: Mitalipov SM

Mitochondria as a therapeutic target for aging and neurodegenerative diseases.

• DOI: 10.2174/156720511795745401
• 发表时间: 2011-06
• 期刊: Current Alzheimer research
• 影响因子: 2.1
• 作者: Reddy PH;Reddy TP
• 通讯作者: Reddy TP

Abnormal mitochondrial dynamics and synaptic degeneration as early events in Alzheimer's disease: implications to mitochondria-targeted antioxidant therapeutics.

• DOI: 10.1016/j.bbadis.2011.10.011
• 发表时间: 2012-05
• 期刊: Biochimica et biophysica acta
• 作者: Reddy PH;Tripathi R;Troung Q;Tirumala K;Reddy TP;Anekonda V;Shirendeb UP;Calkins MJ;Reddy AP;Mao P;Manczak M
• 通讯作者: Manczak M