CYTOSKELETAL TARGETS OF NEURONAL SIGNALING

描述(改编自申请人的摘要)：本提案涉及 细胞外对神经元细胞骨架蛋白的分子调控 信号。这里提出的研究将集中在监管和 多功能微管相关蛋白MAP2的功能 神经元特异性蛋白定位于树突。在此基础上进行的以往研究 项目建立了MAP2‘S的磷酸化状态是原位调节的 通过神经活动，特别是通过神经递质谷氨酸。 磷酸化可以减弱MAP2与微管的结合。 在体外；然而，对其他MAP2功能的影响似乎是可能的。这个 继续这些研究的目的是为了增进我们对 通过对各种map2目标执行详细分析来实现map2功能 相互作用及其通过磷酸化的调节。适用的条件 通过模式突触活性调节MAP2磷酸化将是 用啮齿类动物脑海马片进行了详细的观察。 将使用磷酸表位特异性抗体来检测对 MAP2原位定位，以切片或培养的神经元为模型 系统。钙调神经磷酸酶在MAP2中发挥重要作用的假说 将使用缺乏主要神经元的转基因小鼠来测试调节 钙调神经磷酸酶的异构体。最后，map2与其他基因相互作用的能力 体内分子，包括肌动蛋白细丝和依赖的环-AMP 蛋白激酶，将使用荧光显微镜进行研究。这些 研究涉及几种疾病，在这些疾病中，发育异常或 神经结构的维护是显而易见的，包括精神健康 精神分裂症等疾病，神经退行性疾病 阿尔茨海默氏症和导致智力迟缓的先天性疾病。

DESCRIPTION (Adapted from applicant's abstract): This proposal addresses the molecular control of neuronal cytoskeletal proteins by extracellular signals. The studies proposed here will focus on the regulation and function of the microtubule-associated protein MAP2, a multifunctional neuron-specific protein localized to dendrites. Previous studies under this project established that MAP2's phosphorylation state is regulated in situ by neural activity, especially via the neurotransmitter glutamate. Phosphorylation was shown to attenuate the binding of MAP2 to microtubules in vitro; however, effects on other MAP2 functions appear likely. The continuation of these studies is designed to advance our understanding of MAP2 function by performing detailed analyses of various MAP2 target interactions and their modulation by phosphorylation. Conditions for regulation of MAP2 phosphorylation via patterned synaptic activity will be examined in detail using rodent brain hippocampal slices. Phosphoepitope-specific antibodies will be used to examine the regulation of MAP2 in situ at defined sites, using slices or cultured neurons as model systems. The hypothesis that calcineurin plays an important role in MAP2 regulation will be tested using transgenic mice lacking the major neuronal isoform of calcineurin. Finally, the ability of MAP2 to interact with other molecules in vivo, including actin filaments and the cyclic-AMP dependent protein kinase, will be explored using fluorescence microscopy. These studies are relevant to several disorders in which abnormal development or maintenance of neural structure is manifest, including mental health diseases such as schizophrenia, neurodegenerative diseases such as Alzheimer's, and congenital disorders causing mental retardation.