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Functional Genetic Screen to Identify Interneurons Governing Behaviorally Distinct Aspects of Drosophila Larval Motor Programs

机译:功能基因筛选,以识别控制果蝇幼虫运动程序的行为不同方面的中间神经元。

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Drosophila larval crawling is an attractive system to study rhythmic motor output at the level of animal behavior. Larval crawling consists of waves of muscle contractions generating forward or reverse locomotion. In addition, larvae undergo additional behaviors, including head casts, turning, and feeding. It is likely that some neurons ( e.g. , motor neurons) are used in all these behaviors, but the identity (or even existence) of neurons dedicated to specific aspects of behavior is unclear. To identify neurons that regulate specific aspects of larval locomotion, we performed a genetic screen to identify neurons that, when activated, could elicit distinct motor programs. We used 165 Janelia CRM-Gal4 lines—chosen for sparse neuronal expression—to ectopically express the warmth-inducible neuronal activator TrpA1, and screened for locomotor defects. The primary screen measured forward locomotion velocity, and we identified 63 lines that had locomotion velocities significantly slower than controls following TrpA1 activation (28°). A secondary screen was performed on these lines, revealing multiple discrete behavioral phenotypes, including slow forward locomotion, excessive reverse locomotion, excessive turning, excessive feeding, immobile, rigid paralysis, and delayed paralysis. While many of the Gal4 lines had motor, sensory, or muscle expression that may account for some or all of the phenotype, some lines showed specific expression in a sparse pattern of interneurons. Our results show that distinct motor programs utilize distinct subsets of interneurons, and provide an entry point for characterizing interneurons governing different elements of the larval motor program.
机译:果蝇幼虫爬行是一种有吸引力的系统,可以在动物行为水平上研究节律性运动输出。幼虫爬行由产生向前或向后运动的肌肉收缩波组成。此外,幼虫还具有其他行为,包括头部石膏,转身和进食。可能在所有这些行为中都使用了一些神经元(例如运动神经元),但尚不清楚专门用于行为特定方面的神经元的身份(甚至存在)。为了识别调节幼虫运动特定方面的神经元,我们进行了基因筛选,以识别激活时可引发不同运动程序的神经元。我们使用了165个Janelia CRM-Gal4细胞系(选择用于稀疏的神经元表达)来异位表达温暖诱导型神经元激活剂TrpA1,并筛选了运动缺陷。主屏幕测量了向前的运动速度,我们确定了63条线,其运动速度明显低于激活TrpA1后的对照(28°)。在这些生产线上进行了二次筛选,显示出多种离散的行为表型,包括缓慢的向前运动,过度的反向运动,过度的转向,过度的进食,固定,僵硬的麻痹和延迟性麻痹。尽管许多Gal4品系的运动,感觉或肌肉表达可能解释了部分或全部表型,但有些品系却以稀疏的中间神经元模式表现出特异性表达。我们的结果表明,不同的运动程序利用不同的中间神经元子集,并为表征控制幼虫运动程序中不同元素的中间神经元提供了切入点。

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