LOW-LEVEL LEAD EFFECTS IN GLIAL/ENDOTHELIAL CO-CULTURES

低铅对发育中的神经系统的毒性作用仍然存在 这是这个国家的一个主要健康问题。尚未解决的问题包括 细胞损伤机制，以及铅吸收的机制， 积累铅的脑细胞的储存和耐受性。这个 应聘者的直接职业目标是获得深度和经验 在神经科学领域，特别是星形胶质细胞及其 与血脑屏障(BBB)的相互作用。我们计划彻底地 低水平铅神经毒性的多步骤模型探索 在此应用程序中提供。调查人员提议研究 星形胶质细胞和血管内皮细胞发生的分子变化 培养，在暴露于低水平的铅(0.01至1微米)后。 研究的参数将是钙离子稳态，线粒体膜 电势和间隙结调节。还原型谷氨酰胺 星形胶质细胞的合成酶活性也将作为一个指标进行测量。 铅引起的毒性。调查人员还将检验这一假设 内皮细胞诱导星形胶质细胞成熟并抵抗 共培养中的Pb2+。这位候选人将利用强大的新政策 交互式激光细胞测量技术用于探测细胞的破坏 离散部位的细胞新陈代谢。候选人所在的部门是独一无二的 因为它既有最先进的设备，也有训练有素的 有效使用这项技术所需的支持人员。 交互式激光细胞术和生化分析将结合在一起 评估暴露后这些细胞中发生的分子事件 到亚微摩尔的铅浓度。此外，还建立了一个体外模型。 血脑屏障(BBB)，由大鼠星形胶质细胞和脑组成 毛细血管内皮细胞生长在半透明玻璃的相对表面 可渗透膜，将根据以下方面进行构建和评估 低水平的铅中毒。铅的输运通过由 将测量细胞，以及铅对屏障完整性的影响。 这些实验构成了研究计划的第一阶段，将 获得她的博士学位。最后，在第二阶段，候选人计划 具体解决铅如何进入电池的问题(特别是 星形胶质细胞和内皮细胞)，以及它是如何储存的。候选人 将由一位经验丰富的生物化学家指导这项工作 在蛋白质-金属相互作用以及过渡金属化学中 和新陈代谢。公益广告会解除她的其他责任， 特别是教书，这样她就可以把90%的时间投入到培训中 和研究。候选人有两个长期的职业目标，那就是 作为一名学术科学家和教师，她计划达到：第一，使 在开发有效的体外模型方面取得了重大进展 BBB，然后应用该模型来研究毒素诱导的以及 感染性神经病理学。

The toxic effects of low lead upon the developing nervous system remain a major health issue in this country. Among the unsolved problems are mechanisms of cellular injury, as well as mechanisms of Pb uptake, storage, and tolerance in brain cells that accumulate Pb. The candidate's immediate career objective is to gain depth and experience in the field of neuroscience, specifically with astroglia and their interaction with the blood-brain barrier (BBB). We plans to thoroughly explore the multistep model of low-level lead (Pb) neurotoxicity presented in this application. The investigators propose to study the molecular changes that occur in astroglia and endothelial cells in culture, following exposure to low levels of Pb (0.01 to 1 microM). Parameters investigated will be Ca2+ homeostasis, mitochondrial membrane potential, and gap junction regulation. Reduction of glutamine synthetase activity in astroglia will also be measured as an indicator of Pb-induced toxicity. The investigators will also test the hypothesis that endothelial cells induce astroglial maturation and resistance to Pb2+ in co-culture. The candidate will exploit the powerful new technology of Interactive Laser Cytometry for probing the disruption of cell metabolism at discrete sites. the candidate's department is unique in that it has both state-of-the-art equipment and a highly trained support staff required for effective use of this technology. Interactive Laser Cytometry and biochemical assays will be combined to evaluate the molecular events transpiring in these cells after exposure to submicromolar Pb concentrations. In addition, an in vitro model of the blood-brain barrier (BBB), consisting of rat astroglial and brain capillary endothelial cells grown on opposite surfaces of a semi- permeable membrane, will be constructed and evaluated with respect to low-level Pb toxicity. Pb transport across the barrier formed by the cells will be measured, as well as Pb effects on barrier integrity. These experiments, which constitute Phase I of the research plan, will lead to her PhD degree. Finally, in Phase II, the candidate plans to specifically address the question of how lead enters cells (particularly astroglia and endothelial cells), and how it is stored. The candidate will be guided in this effort by a biochemist with extensive experience in protein-metal interactions, as well as transitional metal chemistry and metabolism. The PSA would relieve her of other responsibilities, particularly teaching, so she could commit 90% of her time to training and research. The candidate has two long-term career objectives, which she plans to attain as an academic scientist and teacher: first, to make significant strides in the development of a valid in vitro model of the BBB, and secondly to apply this model to study toxin-induced as well as infectious neuropathologies.