Molecular basis of glutamate receptor field formation

谷氨酸受体场形成的分子基础

• 批准号: 6700717
• 负责人: DAVID E FEATHERSTONE
• 金额: $ 32.6万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-02-01 至 2007-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6700717
• 关键词: Drosophilidae; arthropod genetics; developmental genetics; developmental neurobiology; gene expression; gene mutation; genetic transcription; genetic translation; glutamate receptor; green fluorescent proteins; messenger RNA; mutant; neurogenesis; neurogenetics; protein structure function; synapses; synaptogenesis

DESCRIPTION (provided by applicant): Excitatory signaling in the central nervous system occurs primarily via glutamate receptors. A molecular understanding of the development, maintenance, and modulation of postsynaptic glutamate receptor fields is therefore very important. However, the molecular mechanisms underlying glutamate receptor field formation are incompletely understood. We are addressing this problem by identifying genes and mechanisms involved in glutamate receptor field formation using a genetic approach in Drosophila. We have isolated several allelic mutations that result in complete and specific loss of postsynaptic glutamate receptor fields. We will identify and clone the mutant gene (which we named 'bad reception', abbreviated 'brec'), then determine the expression and localization of both the brec gene and protein [aim 1]. Properly localized postsynaptic glutamate receptor fields do not form unless the postsynaptic cell is first contacted by the presynaptic cell. After presynaptic contact, glutamate receptor field function increases several hundred-fold within minutes. The molecular mechanisms underlying this process are unknown. To determine whether brec or similar proteins could be involved in this process, we will test whether the initial induction of glutamate receptor fields requires transcription of new receptor subunit genes, or primarily involves translation of pre-existing, possibly pre-localized, receptor subunit mRNAs [aim 2]. To help understand the function of brec and similar proteins in vivo, we will generate and utilize glutamate receptor subunit transgenes tagged with green fluorescent protein (GFP) [aim 3]. Finally, we will continue our forward genetic search for new genes involved in the development of postsynaptic glutamate receptor fields [aim 4] using the new tools and knowledge generated in aims 1-3. The proteins and mechanisms identified in this study (including brec) represent potential drug targets that could be important for understanding and treating neurological conditions such as epilepsy, schizophrenia, damage due to stroke, learning disorders, spinal cord regeneration, or drug addiction

描述（由申请人提供）：中枢神经系统中的兴奋性信号传导主要通过谷氨酸受体发生。因此，对突触后谷氨酸受体区域的发育、维持和调节的分子理解非常重要。然而，谷氨酸受体场形成的分子机制尚不完全清楚。我们正在通过利用遗传方法在果蝇中鉴定参与谷氨酸受体场形成的基因和机制来解决这个问题。我们已经分离出几种等位基因突变，这些突变导致突触后谷氨酸受体区域完全且特异性丧失。我们将识别并克隆突变基因（我们将其命名为“badreception”，缩写为“brec”），然后确定 brec 基因和蛋白质的表达和定位 [目标 1]。除非突触后细胞首先与突触前细胞接触，否则不会形成正确定位的突触后谷氨酸受体区域。 突触前接触后，谷氨酸受体场功能在几分钟内增加数百倍。这一过程背后的分子机制尚不清楚。为了确定 brec 或类似蛋白是否参与这一过程，我们将测试谷氨酸受体区域的初始诱导是否需要新受体亚基基因的转录，或者主要涉及预先存在的、可能预先定位的受体亚基 mRNA 的翻译[目标 2]。为了帮助了解 brec 和类似蛋白质在体内的功能，我们将生成并利用用绿色荧光蛋白 (GFP) 标记的谷氨酸受体亚基转基因 [目标 3]。最后，我们将利用目标 1-3 中产生的新工具和知识，继续对参与突触后谷氨酸受体域发育的新基因进行正向遗传搜索 [目标 4]。本研究中确定的蛋白质和机制（包括 brec）代表了潜在的药物靶点，对于理解和治疗癫痫、精神分裂症、中风造成的损伤、学习障碍、脊髓再生或药物成瘾等神经系统疾病可能很重要