Nutritional Copper Status and the Nervous System

描述(申请人提供)：围产期铜(铜)不足会对中枢神经系统造成重大影响，导致严重的长期神经化学和行为后果。这项研究的长期目标是确定铜的神经化学作用，以更好地了解铜缺乏神经病理的难以捉摸的机制。对啮齿动物的研究表明，铜缺乏会导致特定的局部神经化学变化，围产期铜缺乏后脑铜的恢复可能很困难，如果不是不可能的话。此外，即使在营养补充了几个月后，补充铜的大鼠的感觉-运动功能也发生了变化。人类妊娠和哺乳期的铜RDA可能设定得太低。三个特定的目标是这项研究的重点，将利用Holzman大鼠和小鼠的营养和遗传模型。目的1：我们将建立阻止永久性行为改变和神经病理表达所必需的临界膳食铜水平。我们将确定对大鼠施加不同膳食铜摄入量后的生化和行为结果。还将使用饮食和铜转运蛋白基因Ctrl+/-杂合子的小鼠来研究边缘铜状态。目标2：我们将检验两个具体的假设。使用L-3，4-二羟基苯丝氨酸(L-DOPS)绕过铜依赖酶多巴胺T3-单氧基酶，我们将测试脑内去甲肾上腺素水平降低是否导致大脑发育和运动功能改变。我们将利用生化和行为结果，通过比较铜缺乏对SOD-/-小鼠和野生型对照组的挑战来确定脑中铜，锌-超氧化物歧化酶的作用。目的3：我们将描述缺铜和补铜脑的基因组、蛋白质组和代谢组学特征。大鼠的大脑记录将使用DNA微阵列进行比较。小脑蛋白质图谱将使用二维凝胶电泳法和质谱仪进行比较。大脑代谢物图谱将使用MS和高效液相技术进行比较。

DESCRIPTION (provided by applicant): Insufficient copper (Cu) during perinatal development has a major impact on the central nervous system leading to severe long-term neurochemical and behavioral consequences. The long-range goal of this research is to identify the neurochemical roles of Cu to better understand the elusive mechanisms of Cu-deficient neuropathology. Research in rodents demonstrates that there are selected regional neurochemical alterations induced by Cu deficiency and that restoration of brain Cu following perinatal deficiency may be difficult, if not impossible. Furthermore sensory-motor function is altered in Cu-repleted rats even after months of nutritional supplementation. Human Cu RDAs for pregnancy and lactation may be set too low. Three specific aims are the focus of this research and will utilize nutritional and genetic models with Holzman rats and mice. AIM 1: We will establish the critical dietary Cu level necessary to block expression of permanent behavior alterations and neuropathology. We will determine the biochemical and behavioral outcomes following the imposition of variable dietary copper intakes to rats. Marginal Cu status will also be studied using diet and mice heterozygous for the Cu transporter gene Ctrl +/-. AIM 2: We will test two specific hypotheses. Using L-3,4-dihydroxyphenylserine (L-DOPS) to bypass the Cu-dependent enzyme dopamine t3-monooxvqenase we will test if low brain norepinephrine is responsible for altered brain development and motor function. We will determine the role of brain Cu, Zn-superoxide dismutase by comparing the challenge of Cu deficiency in SOD -/- mice to wild-type controls using biochemical and behavioral outcomes. AIM 3: We will characterize the genomic, proteomic, and metabolomic profile of Cu-deficient and Cu-repleted brain. Rat brain transcripts will be compared using DNA microarrays. Cerebellar protein profiles will be compared using 2-D gel electrophoresis and mass spectrometry. Brain metabolite profiles will be compared using MS and HPLC technology.