AGING AND BRAIN ACETYLCHOLINE RELEASE

这种竞争性续约申请的持续目标是 阐明衰老如何导致大脑中明确的缺陷 神经递质释放，集中在胆碱能系统。 作为 我们在生物化学水平上进一步探讨这个问题， 提供了有关监管的见解， 正常AS神经递质释放过程及其调节 以及衰老的动物。 未来四年的重点将从钙- 通量和效力一般的调节作用， 蛋白激酶C在这些释放缺陷。 我们的结果和 其他人认为这种酶是一个节点的影响 衰老和膜过氧化作用对递质释放的影响。 具体而言，我们建议回答以下问题： 1. 衰老是否会影响蛋白激酶C的活性， 大脑区域和准备类似吗？ 2. 衰老是如何干扰佛波酯诱导的 乙酰胆碱（ACh）的释放和蛋白质的移位 激酶C？ 3. 老化是否会影响钙离子载体的效力， 与蛋白激酶C易位有关吗 4. 将蛋白激酶C直接注入突触体 影响蛋白质磷酸化和乙酰胆碱释放？ 5. 衰老是否影响蛋白激酶C介导的 突触体中蛋白质的原位磷酸化 6. 磷脂酰丝氨酸是否能恢复老年人的年龄相关缺陷？ 蛋白激酶C易位正常？ 7. 外源性突触体膜过氧化作用是否模拟了 衰老对蛋白激酶C活性的影响， 释放和膜流动性？ 这些研究结合了联合收割机的各种酶测定和 在我的实验室中使用的药理学方法，沿着 适于注入酶或其它的病毒外壳注入方法 大分子进入神经末梢。 我们将继续使用 稀释的突触体作为选择的原因 如前所述。

The continuing goal of this competitive renewal application is to elucidate how aging causes well defined deficits in brain neurotransmitter release, focusing on the cholinergic system. As we explore this problem further at the biochemical level, new insights are provided about the regulation of the neurotransmitter-release process and its modulation in normal as well as senescent animals. The focus of the next four years shifts from that of calcium- fluxes and potency generally to that of the modulatory role of protein kinase C in these release-deficits. Our results and others suggest that this enzyme is a nodal point for the effects of aging and membrane peroxidation on transmitter release. Specifically, we propose to answer the following questions: 1. Does aging affect protein kinase C activity in different brain regions and preparations similarly? 2. How does aging interfere with the phorbol ester-induced release of acetylcholine (ACh) and the translocation of protein kinase C? 3. Does aging affect the potency of calcium ionophores with respect to protein kinase C translocation? 4. Does infusion of protein kinase C directly into synaptosomes affect protein phosphorylation and ACh release? 5. Does aging affect the protein kinase C-mediated phosphorylation of proteins in synaptosomes in situ? 6. Does phosphatidylserine restore age-related deficits in protein kinase C translocation to normal? 7. Does exogenous synaptosomal membrane peroxidation mimic the effects of aging on protein kinase C activities as it does ACh release and membrane fluidity? These studies combine a variety of enzyme assays and pharmacological approaches used in my laboratory, along with a viral coat infusion procedure adapted to infuse enzymes or other macromolecules into nerve terminals. We will continue to use diluted synaptosomes as the preparation of choice for reasons described in the previous grant.