SYNAPTIC AND EXTRASYNAPTIC NMDA RECEPTORS AND NEUROPROTECTIVE LIPID SIGNALING

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 具体目标 1.目的：利用15-脂氧合酶（15-lipoxygenase，15-LOX）开放阅读框（open reading frame，ORF）和小干扰RNA（small-interfering RNA，siRNA）质粒分别转染原代培养的大鼠海马神经元，使其过表达和敲低15-LOX。 免疫细胞化学用于检测蛋白表达和神经元活力。 2.使用转染的培养物测定对淀粉样肽（A <$42）的保护作用 诱导毒性。 预期15-LOX的过表达将激活神经保护途径，而siRNA敲低培养物将更容易受到A 42诱导的毒性的影响。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Specific Aims 1. To successfully transfect primary cultures of rat hippocampal neurons with the 15-LOX (15-lipoxygenase) open reading frame (ORF) and siRNA (small-interference RNA) plasmids to over-express and knock-down 15-LOX, respectively. Immunocytochemistry was used to examine for protein expression and neuron viability. 2. To use the transfected cultures to assay for protection against amyloid-¿ peptide (A¿42) induced toxicity. It is expected that over expression of the 15-LOX will activate neuroprotective pathways, whereas the siRNA-knock-down cultures will be more susceptible to A¿42-induced toxicity.