PROTEIN PHOSPHORYLATION AND LTP IN AGED ANIMALS

老年人和阿尔茨海默病患者 通常有严重的认知障碍。年龄相近的动物 在学习和记忆方面也有严重的缺陷。然而， 直到最近，才在以下方面取得重大进展 理解认知的分子和细胞基础 功能正常。从而了解了细胞和分子基础。 与年龄相关的学习缺陷才刚刚开始出现。长期的 增强(LTP)是突触可塑性的一种形式，已经被广泛地 被吹捧为学习和记忆的细胞积木。在.期间 在之前的授权期内，P.I.在LTP诱导方面显示出明显的缺陷 老年Fischer 344只大鼠的CA1区。LTP的分子相关因素有 也是以前工作的一个主要重点。私家侦探已经证明了LTP诱导 刺激导致突触蛋白的磷酸化增加 囊泡相关的磷蛋白，还表明老年动物有 突触素磷酸化的显著缺陷。此外，私家侦探和 他的同事们最近开发出了所需的免疫试剂 研究NMDA和海人藻酸/AMPA受体。这些受体被认为是 在LTP中发挥关键作用，在老年人中似乎有缺陷 动物。在目前的提案中，先前关于年龄性缺乏症的工作 LTP机制的扩展和NMDA和海人藻酸盐/AMPA的新研究 受体是有计划的。其次，私家侦探提议测试 治疗(卡路里限制和抗氧化治疗) 据报道，改善认知和其他随年龄增长的缺陷可能会限制 或消除特定LTP机制中与年龄相关的缺陷。这些 研究将涉及细胞和分子分析。具体的 重点将放在四个领域。首先，私家侦探会检验这一假设 老年动物在齿状回和CA3区的LTP诱导方面将出现缺陷 海马体的区域。然后，私家侦探将检验这一假设 抗氧化剂治疗和/或限制热量摄入可能会延缓衰老 LTP诱导相关缺陷。其次，私家侦探提议测试 老年动物交感蛋白磷酸化缺陷的假说 由电生理刺激引起的。当赤字被发现时，它就是 建议测试抗氧化治疗和/或卡路里限制 因为他们有能力改善这些赤字。私家侦探建议测试 假设NMDA和/或海人藻酸/AMPA受体功能、表达 和/或磷酸化在老年动物中发生改变。如果与年龄相关 如果选择了这两个受体中的任何一个的缺陷，P.I.将测试 卡路里限制和/或抗氧化剂治疗可能 改善这些赤字。在这项提案的最终目标中，P.I. 将检验这样一种假设，即LTP诱导后的激酶激活可能 缺乏古老的动物。此外，私家侦探还提出， 与项目1、2和4合作。纯化的激酶和抗体将 为GABAA-R和钙离子通道功能的合作研究提供依据 分别与项目1和项目2进行协作实验。最后， 将检测突触素I的表达作为移植的衡量标准 与项目4合作开发。

Aged individuals and individuals suffering from Alzheimer's disease typically have significant cognitive impairment. Similarly aged animals are also known to have profound deficits in learning and memory. However, it is only recently that significant progress has been made in understanding the molecular and cellular substrates of cognitive functioning. Thus an understanding of the cellular and molecular bases of age related-learning deficits is just beginning to emerge. Long-term potentiation (LTP) is a form of synaptic plasticity that has been widely touted as a cellular building block of learning and memory. During the previous grant period the P.I. demonstrated clear deficits in LTP induction in the CA1 area of aged Fischer 344 rats. Molecular correlates of LTP were also a major focus of previous work. The P.I. has shown that LTP-inducing stimuli produce an increase in the phosphorylation of synapsin a synaptic vesicle-associated phosphoprotein and has also shown that aged animals have significant deficits in synapsin phosphorylation. Moreover, The P.I. and his colleagues have recently developed the immunological reagents required to study the NMDA and kainate/AMPA receptors. These receptors are thought to play critical role in LTP and they appear to be defective in old animals. in the current proposal previous work on age-related deficits in LTP mechanisms will be extended and new studies of NMDA and kainate/AMPA receptors are planned. Second, the P.I. proposes to test whether treatments (caloric restriction an antioxidant therapy) which have been reported to ameliorate cognitive and other deficits seen with age may limit or eliminate age-related deficits in specific LTP mechanisms. These studies will involve both cellular and molecular analyses. The specific foci will be in four areas. First, the P.I. will test the hypothesis that aged animals will exhibit deficits in LTP induction in the dentate and CA3 regions of the hippocampus. The P.I. will then test the hypothesis that antioxidant treatment and/or caloric restriction may ameliorate age related-deficits in LTP induction. Second, the P.I. proposes to test the hypothesis that aged animals have deficits in symapsin phosphorylation induced by electrophysiological stimulation. When deficits are found it is proposed that antioxidant treatment and/or caloric restriction be tested for their ability to ameliorate these deficits. The P.I. proposes to test the hypothesis that NMDA and/or kainate/AMPA receptor function, expression and/or phosphorylation are altered in aged animals. If age related deficits in either of these receptors is selected, the P.I. will test the hypothesis that caloric restriction and/or antioxidant treatment may ameliorate these deficits. In the final aim of this proposal, the P.I. will test the hypothesis that kinase activation following LTP induction may be deficient in old animals. In addition, the P.I. also proposed to collaborate with Projects 1, 2 and 4. Purified kinases and antibodies will be provided for collaborative studies of GABAA-R and Ca2+ channel function in collaborative experiments with Projects 1 and 2 respectively. Lastly, synapsin I expression will be assayed as a measure of transplant development in collaboration with Project #4.