KINASE-MEDIATED SIGNALING PATHWAYS IN NEURONAL APOPTOSIS

神经元凋亡中激酶介导的信号通路

• 批准号: 6639703
• 负责人: MARY LOU VALLANO
• 金额: $ 26.48万
• 依托单位: UPSTATE MEDICAL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-07-01 至 2005-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6639703
• 关键词: apoptosis; biological signal transduction; calcium flux; genetically modified animals; granule cell; laboratory mouse; neurotrophic factors; phosphorylation; protein kinase; tissue /cell culture; transcription factor

DESCRIPTION (From the applicant's abstract): We are studying the kinase-mediated signaling pathways that promote neuronal survival and prevent apoptosis in primary cultures of granule neurons. Relevant neurotrophins are insulin-like growth factor (IGF-1) and brain-derived neurotrophic factor (BDNF). Glutamate is the predominant neurotransmitter that regulates neuronal activity through its interaction with different glutamate receptor-channels, referred to herein as N-methyl-D-aspartate (NMDA) and non-NMDA receptors. Binding of these agents to their respective receptors in the neuronal plasma membrane initiates activation of intracellular signaling cascades that regulate the transcription of pro-apoptotic or anti-apoptotic genes. The balance between these gene products ultimately determines cell fate. Early in granule cell development, before neurons have formed synaptic connections, they rely exclusively on neurotrophic factors, present in the serum that bathes them, for survival. We have novel evidence that particular isoforms of the family of phospholipid-sensitive protein kinases (PKCs), designated as atypical PKCs (aPKCs), are critical components of the neurotrophin-dependent survival pathway. We are quite enthusiastic about these data since almost nothing is currently known about the neuronal functions of aPKCs. Moreover, preliminary data indicate that inhibition of aPKCs reduces phosphorylation on serine-133 of the nuclear transcription factor CREB. One goal of this application is to extend our studies examining the linkage between neurotrophin-dependent activation of aPKCs. phosphorvlation of CREB. and survival. In many neuronal populations, inadequate electrical stimulation by excitatory neurotransmitters or inadequate trophic factor make cell death more likely. A "Ca2+ set-point hypothesis" postulates that the bulk cytosolic Ca2+ activity determines the degree to which immature neurons require trophic factor to suppress the mechanism responsible for apoptosis (Koike et al., 1989). Accordingly, many types of neurons grown in dissociated culture can be rescued from death, induced by neurotrophin deprivation, by inclusion of agents in the media that produce a sustained elevation in Ca2+. These agents include elevated KCI acting through voltage-sensitive Ca2+ channels (VSCCs) and NMDA acting directly upon NMDA receptor channels (NRs). We have novel evidence that a Ca2+ and calmodulin-dependent kinase, designated as CaM KIV, and its substrate CREB are critical components of this Ca2+-dependent signaling pathway. Immunocytochemical evidence also indicated that these treatments induce the redistribution of CaM KIV from the nucleus to the cytosol. A second goal of this application is to extend our studies examining the linkage between calcium, CaM KIV phosphorvlation of CREB, and survival. Phosphorylation and activation of CREB may mediate induction of anti-apoptotic genes, such as bc1-2, or repression of pro-apoptotic genes, such as bax. A third goal in this application is to compare the expression levels of bc1-2 and bax rnRNAs under culture conditions promoting survival or apoptosis.

描述（摘自申请人的摘要）：我们正在研究 激酶介导的信号通路，促进神经元存活并预防 颗粒神经元原发性培养物的凋亡。相关的神经营养蛋白是 胰岛素样生长因子（IGF-1）和脑衍生的神经营养因子 （bdnf）。谷氨酸是调节神经元的主要神经递质 通过与不同的谷氨酸受体通道相互作用的活动， 此处称为N-甲基-D-天冬氨酸（NMDA）和非NMDA受体。 这些药物与神经元血浆中各自受体的结合 膜启动调节细胞内信号传导级联 促凋亡或抗凋亡基因的转录。之间的平衡 这些基因产物最终决定了细胞命运。颗粒细胞的早期 开发，在神经元建立突触连接之前，它们依靠 仅在沐浴它们的血清中存在于神经营养因素上 生存。我们有新的证据表明家族的特殊同工型 磷脂敏感蛋白激酶（PKC），被指定为非典型PKCS （APKC）是神经营养蛋白依赖性生存的关键成分 路径。我们对这些数据非常热情，因为几乎没有 目前知道APKC的神经元功能。而且，初步 数据表明，抑制APKC会降低对丝氨酸133的磷酸化 核转录因子CREB。该应用程序的一个目标是 扩展我们的研究检查神经营养依赖性之间的联系 APKC的激活。 Creb的磷酸化。和生存。在许多神经元中 种群，兴奋性神经递质电刺激不足 或营养因子不足会使细胞死亡更有可能。 “ CA2+设定点 假设“假设大量的胞质Ca2+活性决定了 未成熟神经元需要营养因子以抑制的程度 负责凋亡的机制（Koike等，1989）。因此，许多 可以从死亡中拯救出在解离文化中生长的神经元的类型， 由神经营养素剥夺引起的，通过在培养基中纳入剂 在Ca2+中产生持续的升高。这些代理人包括高高的KCI表演 通过对电压敏感的Ca2+通道（VSCC）和NMDA直接作用 NMDA受体通道（NRS）。我们有新的证据表明CA2+和 钙调蛋白依赖性激酶，被指定为CAM KIV，其底物Creb是 此Ca2+依赖性信号通路的临界组件。 免疫细胞化学证据还表明，这些治疗诱导 CAM KIV从细胞核重新分布。第二个目标 该应用是为了扩展我们的研究，以研究 CREB的钙，CAM KIV磷酸化和存活。磷酸化和 CREB的激活可能介导抗凋亡基因的诱导，例如 BC1-2或促凋亡基因的抑制，例如Bax。这是第三个目标 应用是比较BC1-2和BAX RNRNA的表达水平 培养条件促进生存或凋亡。

项目成果

Roscovitine triggers excitotoxicity in cultured granule neurons by enhancing glutamate release.

Roscovitine 通过增强谷氨酸释放来触发培养的颗粒神经元的兴奋性毒性。

• DOI: 10.1124/mol.105.012732
• 发表时间: 2005
• 期刊: Molecular pharmacology
• 影响因子: 3.6
• 作者: Monaco3rd,EdwardA;Vallano,MaryLou
• 通讯作者: Vallano,MaryLou

Reduced blockade by extracellular Mg(2+) is permissive to NMDA receptor activation in cerebellar granule neurons that model a migratory phenotype.

细胞外 Mg(2 ) 的阻断减少，允许模拟迁移表型的小脑颗粒神经元中 NMDA 受体激活。

• DOI: 10.1111/j.1471-4159.2010.06746.x
• 发表时间: 2010
• 期刊: Journal of neurochemistry
• 影响因子: 4.7
• 作者: Gerber,AdamM;Beaman-Hall,CarolM;Mathur,Anjili;Vallano,MaryLou
• 通讯作者: Vallano,MaryLou

Depolarization and Ca(2+) down regulate CB1 receptors and CB1-mediated signaling in cerebellar granule neurons.

去极化和 Ca(2) 下调小脑颗粒神经元中 CB1 受体和 CB1 介导的信号传导。

• DOI: 10.1016/j.neuropharm.2005.11.012
• 发表时间: 2006
• 期刊: Neuropharmacology
• 影响因子: 4.7
• 作者: Vallano,MaryLou;Beaman-Hall,CarolM;Bui,CuongJ;Middleton,FrankA
• 通讯作者: Middleton,FrankA

Ca2+ and CaM kinase regulate neurofilament expression.

Ca2 和 CaM 激酶调节神经丝表达。

• DOI: 10.1097/00001756-200311140-00013
• 发表时间: 2003
• 期刊: Neuroreport
• 影响因子: 1.7
• 作者: Bui,CuongJ;Beaman-Hall,CarolM;Vallano,MaryL
• 通讯作者: Vallano,MaryL

Transcriptional profiling of depolarization-dependent phenotypic alterations in primary cultures of developing granule neurons.

发育中的颗粒神经元原代培养物中去极化依赖性表型改变的转录谱。

• DOI: 10.1016/j.brainres.2006.08.043
• 发表时间: 2006
• 期刊: Brain research
• 影响因子: 2.9
• 作者: Bui,CuongJ;McGann,AlexandraC;Middleton,FrankA;Beaman-Hall,CarolM;Vallano,MaryLou
• 通讯作者: Vallano,MaryLou