Gene Expression Analysis of Aplysia Neural Network

海兔神经网络基因表达分析

• 批准号: 7424034
• 负责人: JINGYUE JU
• 金额: $ 54.7万
• 依托单位: COLUMBIA UNIV NEW YORK MORNINGSIDE
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-01 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7424034
• 关键词: Address; Affinity; Aftercare; Animal Model; Anti-Sense Probes; Aplysia; Appearance; Behavior; Behavioral; Binding; Biological; Biological Neural Networks; Caenorhabditis elegans; Cell Nucleus; Cell physiology; Cells; Chromosome Pairing; Code; Collection; Communication; Complementary DNA; Complex; Cytoplasmic Protein; Databases; Development; Dissection; Drosophila genus; EST Library; Event; Expressed Sequence Tags; FMRFamide; Facility Construction Funding Category; Family; Figs - dietary; Flowcharts; Future; Ganglia; Gene Expression; Gene Expression Profile; Gene Expression Profiling; Gene Expression Regulation; Gene Transfer; Genes; Genetic; Genome; In Situ Hybridization; In Vitro; Individual; Invertebrates; Kinesin; Large-Scale Sequencing; Lead; Learning; Length; Libraries; Localized; Location; Long-Term Depression; Longitudinal Studies; Maps; Marines; Measurement; Membrane; Memory; Mental Depression; Messenger RNA; Microarray Analysis; Microsomes; Molecular; Molecular Biology; Molecular Profiling; Monitor; Motor Neurons; Myxoid cyst; Nervous system structure; Neuraxis; Neurobiology; Neuronal Plasticity; Neurons; Neurosciences; Neurosciences Research; Neurotransmitters; Organelles; Organism; Pleural; Polyribosomes; Population; Process; Protein Biosynthesis; Protein Export Pathway; Proteins; RNA; RNA Transport; Regulation; Research; Research Personnel; Resources; Reverse Transcriptase Polymerase Chain Reaction; Rough endoplasmic reticulum; Sensory; Serotonin; Site; Source; Synapses; Synaptic Vesicles; Synaptic plasticity; System; Time; Transcript; Translating; Translations; Work; base; cDNA Arrays; cDNA Library; fly; impression; in vivo; long term memory; member; neural circuit; programs; relating to nervous system; size

DESCRIPTION (provided by applicant): Aplysia californica has been one of the dominant invertebrate model organisms utilized for research on the neurobiology of behavior, learning, and long-term memory. A key finding in recent studies is that both the functional and structural changes for memory storage are synapse-specific and require local translation. To understand the molecular mechanisms underlying long-term memory storage including its structural underpinning, it will be essential to determine how the population of mRNAs is destined for translation in synapses during memory formation. To examine this issue, we will extract mRNAs from kinesin transport complexes from ganglia and cultured individual sensory and motor neurons following treatment with the neurotransmitter serotonin (involved in learning-related long-term facilitation) and with FMRFamide (involved in long-term inhibition). At several time points after treatment, we will also characterize the mRNAs being actively translated on free and membrane-bound polysomes. The latter should include proteins targeted to the synaptic vesicles and membranes. Isolated mRNAs from all 3 populations (kinesin complexes, free polysomes, bound polysomes) will be hybridized to an Aplysia cDNA microarray containing features specific for the majority of central nervous system (CNS) genes. This microarray, an expansion of our current array, will be constructed from the unique clones in the EST libraries we have already generated from individual neurons, pedal-pleural ganglia, and the whole CNS of Aplysia. While we estimate these clones cover nearly 80% of all genes expressed in Aplysia neurons, only a fraction of the library has been annotated due to insufficient coding information. Nearly full annotation of the libraries will be achieved by sequencing full-length cDNAs from the same CNS sources. The full-length cDNA libraries and the Aplysia microarray will provide important resources for investigators to study learning and other behavior paradigms in this important model organism.

描述（由申请人提供）：海兔是用于研究行为、学习和长期记忆的神经生物学的主要无脊椎动物模型生物之一。最近研究的一个重要发现是，记忆存储的功能和结构变化都是突触特异性的，并且需要局部翻译。为了了解长期记忆存储的分子机制，包括其结构基础，有必要确定记忆形成过程中 mRNA 群体如何在突触中进行翻译。为了研究这个问题，我们将从神经节的驱动蛋白转运复合物中提取 mRNA，并在用神经递质血清素（涉及与学习相关的长期促进）和 FMRFamide（涉及长期抑制）治疗后培养单个感觉和运动神经元。 。在治疗后的几个时间点，我们还将表征在游离和膜结合多核糖体上主动翻译的 mRNA。后者应包括针对突触小泡和膜的蛋白质。来自所有 3 个群体（驱动蛋白复合物、游离多核糖体、结合多核糖体）的分离 mRNA 将与含有大多数中枢神经系统 (CNS) 基因特异性特征的海兔 cDNA 微阵列杂交。该微阵列是我们当前阵列的扩展，将由我们已经从单个神经元、足胸膜神经节和海兔的整个中枢神经系统生成的 EST 文库中的独特克隆构建。虽然我们估计这些克隆覆盖了海兔神经元中表达的所有基因的近 80%，但由于编码信息不足，仅对文库的一小部分进行了注释。 通过对来自相同 CNS 来源的全长 cDNA 进行测序，可以实现对文库的近乎完整的注释。全长 cDNA 文库和海兔微阵列将为研究人员研究这一重要模式生物的学习和其他行为范式提供重要资源。