Department of Biological Sciences, Vanderbilt University and Medical Center.
Department of Biological Sciences, Vanderbilt University and Medical Center; Department of Cell and Developmental Biology, Vanderbilt University and Medical Center; Department of Pharmacology, Vanderbilt University and Medical Center; Kennedy Center for Research on Human Development, Vanderbilt University and Medical Center; Vanderbilt Brain Institute, Vanderbilt University and Medical Center;
J Vis Exp. 2024 Mar 1(205). doi: 10.3791/66629.
Early-life olfactory sensory experience induces dramatic synaptic glomeruli remodeling in the Drosophila juvenile brain, which is experientially dose-dependent, temporally restricted, and transiently reversible only in a short, well-defined critical period. The directionality of brain circuit synaptic connectivity remodeling is determined by the specific odorant acting on the respondent receptor class of olfactory sensory neurons. In general, each neuron class expresses only a single odorant receptor and innervates a single olfactory synaptic glomerulus. In the Drosophila genetic model, the full array of olfactory glomeruli has been precisely mapped by odorant responsiveness and behavioral output. Ethyl butyrate (EB) odorant activates Or42a receptor neurons innervating the VM7 glomerulus. During the early-life critical period, EB experience drives dose-dependent synapse elimination in the Or42a olfactory sensory neurons. Timed periods of dosed EB odorant exposure allow investigation of experience-dependent circuit connectivity pruning in juvenile brain. Confocal microscopy imaging of antennal lobe synaptic glomeruli is done with Or42a receptor-driven transgenic markers that provide quantification of synapse number and innervation volume. The sophisticated Drosophila genetic toolkit enables the systematic dissection of the cellular and molecular mechanisms mediating brain circuit remodeling.
早期嗅觉感觉体验会在果蝇幼脑中引起剧烈的突触小球重塑,这种重塑与体验剂量有关,在短暂的、明确的关键期内具有时间限制且是暂时可逆的。大脑回路突触连接重塑的方向性取决于作用于应答受体类嗅觉感觉神经元的特定气味剂。一般来说,每个神经元类仅表达单个气味受体,并支配单个嗅觉突触小球。在果蝇遗传模型中,通过气味反应性和行为输出,已经精确地绘制出了全部嗅觉小球图谱。丁酸乙酯(EB)气味剂激活支配 VM7 小球的 Or42a 受体神经元。在生命早期的关键期内,EB 体验会导致 Or42a 嗅觉感觉神经元中依赖剂量的突触消除。定时的、剂量的 EB 气味暴露允许研究幼脑中经验依赖性的回路连接修剪。利用 Or42a 受体驱动的转基因标记物对触角叶突触小球进行共聚焦显微镜成像,提供突触数量和神经支配体积的定量分析。复杂的果蝇遗传工具包使系统剖析介导大脑回路重塑的细胞和分子机制成为可能。