Engrailed and the Control of Synaptic Circuitry in Drosophila

果蝇的纠缠和突触回路的控制

• 批准号: 7683182
• 负责人: JONATHAN M BLAGBURN
• 金额: $ 11.25万
• 依托单位: UNIVERSITY OF PUERTO RICO MED SCIENCES
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-15 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7683182
• 关键词: Afferent Neurons; Animals; Auditory; Autistic Disorder; Binding; Biological Models; Brain; Branchiostoma floridae AmphiEn protein; Cell surface; Cells; Cereals; Complex; Confocal Microscopy; Data; Development; Dictyoptera; Disease; Drosophila genus; Drosophila melanogaster; Ectopic Expression; Evolution; Florida; Genes; Genetic; Genetic Models; Genetic Screening; Genetic Transcription; Green Fluorescent Proteins; Health; Human; Institutes; Knockout Mice; Learning; Link; Membrane; Mentors; Midbrain structure; Modeling; Molecular; Molecular Biology; Morphology; Nervous system structure; Neurons; Parkinson Disease; Pathway interactions; Periplaneta americana; Pilot Projects; Play; Process; Research; Role; Sensory; Specificity; Symptoms; Synapses; System; Techniques; Testing; Time; Touch sensation; Universities; Work; autism spectrum disorder; axonal guidance; cofactor; dopaminergic neuron; genetic manipulation; nervous system disorder; neural circuit; neuronal survival; retinal rods; sensory system; synaptogenesis

DESCRIPTION (provided by applicant): Engrailed is a ubiquitous transcriptional regulator that is potentially of great significance to human health. It has recently been linked to fate determination and survival of midbrain dopaminergic neurons, with En knockout mice showing Parkinson-like symptoms, and has also been linked to autism spectrum disorder. It is therefore essential to understand the role En plays in regulating neuronal connectivity. The long term objectives of this research are to use simple model systems, that have identifiable neurons, to investigate how Engrailed controls synaptic target recognition, and what are the downstream effector genes that it regulates. My previous work has used the cockroach cereal system to study this; however, there are substantial barriers to further progress in this system. The aims of this pilot project will enable me to develop a similar system of identifiable Engrailed-expressing neurons in the genetically tractable Drosophila melanogaster. The first aim is to use Gal4-UAS to express green fluorescent protein in neurons that normally express Engrailed. Confocal microscopy will then be used to characterize these neurons in a range of sensory systems, such as the olfactory, auditory and touch cells. The second aim is to focus on one of these sensory systems using the electrophysiological and anatomical techniques that have been developed in the lab, and the expertise of the mentor, Dr. Rod Murphey, in order to be able to test the role of Engrailed in controlling the synaptic connectivity of this system. The third aim will involve the co-expression of ectopic Engrailed along with GFP, enabling a direct test of the idea that Engrailed controls connectivity in the circuit. In the final aim, with the help of Dr. Rod Murphey, I will use genetic screens to begin a search for other genes downstream of Engrailed that control synaptic specificity. An alternative strategy will be to test En binding targets previously identified by other groups. Drosophila models are particularly useful for the discovery of molecular pathways that are directly relevant to human health, because most of these pathways have been conserved during evolution. All animals have Engrailed protein, so it is very likely that any molecules that are regulated by it during the process of synapse formation in Drosophila have their counterparts in humans. These molecules may be of great potential importance in neurological diseases such as Parkinson's or autism.

描述（由申请人提供）：Engrailed是一种普遍存在的转录调节因子，对人类健康可能具有重要意义。它最近与中脑多巴胺能神经元的命运决定和存活有关，En敲除小鼠表现出帕金森样症状，并且还与自闭症谱系障碍有关。因此，了解En在调节神经元连接中的作用至关重要。本研究的长期目标是使用具有可识别神经元的简单模型系统来研究Engrailed如何控制突触目标识别，以及它调节的下游效应基因是什么。我以前的工作已经使用蟑螂谷物系统来研究这一点;然而，在这个系统中进一步取得进展存在实质性障碍。这个试验项目的目的将使我能够在遗传上易处理的果蝇中开发一个类似的可识别的表达Engrailed的神经元系统。第一个目的是使用Gal 4-UAS在正常表达Engrailed的神经元中表达绿色荧光蛋白。然后，共聚焦显微镜将用于表征这些神经元在一系列的感觉系统，如嗅觉，听觉和触觉细胞。第二个目标是使用实验室开发的电生理和解剖技术以及导师Rod Murphey博士的专业知识来关注这些感觉系统之一，以便能够测试Engrailed在控制该系统的突触连接方面的作用。第三个目标将涉及异位Engrailed沿着与GFP的共表达，从而能够直接测试Engrailed控制电路中的连接性的想法。在最后一个目标中，在罗德·墨菲博士的帮助下，我将使用基因筛选来开始寻找控制突触特异性的其他基因。另一种策略是测试先前由其他组鉴定的En结合靶点。 果蝇模型对于发现与人类健康直接相关的分子通路特别有用，因为这些通路中的大多数在进化过程中是保守的。所有的动物都有Engrailed蛋白，所以很可能在果蝇突触形成过程中受其调节的任何分子在人类中都有对应物。这些分子可能在神经系统疾病如帕金森氏症或自闭症中具有巨大的潜在重要性。