NEURONAL CA++ SEQUESTERING COMPARTMENTS PROTECTING ROLE

该补助金的主要研究重点是深入了解 细胞内钙离子螯合的结构和功能 神经元的间隔。 这些隔间非常有趣 因为最近的证据表明它们在保护 在暴露于谷氨酸的神经元中，谷氨酸是主要的兴奋性神经递质 和“兴奋性毒性”的原因。 兴奋性毒性是一种主要的 中风和创伤中神经元损伤的原因以及可能的 在与年龄相关的神经变性疾病中神经元损伤的介质， 例如亨廷顿舞蹈症渐冻症 具体目标是：1.定位CaATP酶的特异性亚型 和钙网蛋白2，通过神经元钙离子螯合成分 在大脑中的隔室和培养的神经元和神经母细胞瘤细胞。 2. 为了研究CaATPase和 钙网蛋白随后到达Ca 2+螯合隔室。 3.到 研究提高胞浆Ca 2+的试剂对mRNA的影响， 使用神经母细胞瘤细胞的CaATP酶和钙网蛋白的蛋白水平， 培养的神经元 4.用cDNA转染神经母细胞瘤细胞， CaATP酶和钙网蛋白，以确定这些蛋白质是否具有保护作用 对抗升高的胞质Ca 2+和5.雇用 细胞内染料，以直接测量神经元中的细胞溶质Ca2+， 神经母细胞瘤细胞 我们将采用生物化学，形态学和 分子技术在这些研究中的应用。

The principle research focus of this grant is to gain insight into the structure and function of the intracellular Ca2+ sequestering compartments of neurons. These compartments are of great interest because recent evidence has accumulated that they play a protective role in neurons exposed to glutamate, the major excitatory neurotransmitter in the CNS and the cause of "excitotoxicity". Excitotoxicity is a major cause of neuronal damage in stroke and trauma as well as a likely mediator of neuronal injury in age related neurodegenerative diseases, e.g. Huntington's, ALS. The specific aims are: 1. To localize the specific isoforms of CaATPase and calreticulin two, by components of neuronal Ca2+ sequestering compartments in brain and cultured neurons and neuroblastoma cells. 2. To investigate the biosynthetic route(s) by which the CaATPase and calreticulin follow to reach the Ca2+ sequestering compartments. 3. To study the effects of agents which raise cytosolic Ca2+ on the mRNA and protein level of CaATPase and calreticulin using neuroblastoma cells and cultured neurons. 4. To transfect neuroblastoma cells with cDNAs for CaATPase and calreticulin to determine if these proteins are protective against toxic effects of elevated cytosolic Ca2+ and 5. To employ intracellular dyes to directly measure cytosolic Ca2+ in neurons and neuroblastoma cells. We will employ biochemical, morphological and molecular techniques in these investigations.