Department of Pediatrics, School of Medicine, University of California, San Diego, La Jolla, California.
Section of Cell and Developmental Biology, Division of Biological Sciences, University of California, San Diego, La Jolla, California.
Curr Protoc. 2022 Oct;2(10):e576. doi: 10.1002/cpz1.576.
Neurodevelopmental disorders are a heterogeneous group of behaviorally defined disorders with both genetic and environmental risk factors. Given that many neurodevelopmental disorders are characterized by impaired learning and/or intellectual abilities, behavioral paradigms that assess cognition in animal models have been effective tools in delineating underlying genetic variants that impact disease pathophysiology. For example, learning and memory paradigms in the common fruit fly Drosophila melanogaster have been successfully used to study risk genes and biological pathways associated with several neurodevelopmental disorders, including fragile X syndrome, autism spectrum disorder, and CHARGE syndrome. While these established Drosophila behavioral paradigms have historically been used to investigate genetic risk factors, they also have many other applications, including use in developmental neurotoxicology studies to determine environmental risk factors for neurodevelopmental disorders. There is, however, a deficit of step-by-step protocols that describe how to apply learning and memory assays in fruit flies to developmental neurotoxicology studies. Here, we describe two quantitative behavioral paradigms for Drosophila-predator-induced oviposition and courtship conditioning-that can be used to measure different forms of learning and memory in the context of a developmental neurotoxicology study. Non-associative learning and memory are measured here by examining female Drosophila oviposition behavior in response to endoparasitoid wasps, while associative learning and memory are measured in males using courtship conditioning. Our protocols outline procedures for oral toxicant exposure of developing fruit flies, culturing of endoparasitoid wasps, measuring Drosophila oviposition activity, and assessing conditioned courtship in order to identify the impact of toxicants on learning and memory in both females and males. As an example, we present the protocols using bisphenol A, a chemical utilized in the synthesis of polycarbonate plastics, to determine its impacts on learning and memory. These protocols are inexpensive and relatively simple to perform, and can be used by labs that are new to Drosophila behavioral research to evaluate how toxicant exposure influences learning and memory in male and female flies. © 2022 Wiley Periodicals LLC. Basic Protocol 1: Preparation of toxicant-containing food and developmental exposure Basic Protocol 2: Predator-induced oviposition assay Support Protocol: Culture of Leptopilina heterotoma Basic Protocol 3: Conditioned courtship assay.
神经发育障碍是一组具有遗传和环境风险因素的行为定义障碍。鉴于许多神经发育障碍的特征是学习和/或智力能力受损，评估动物模型认知的行为范式已成为确定影响疾病病理生理学的潜在遗传变异的有效工具。例如，在常见的黑腹果蝇 Drosophila melanogaster 中学习和记忆范式已成功用于研究与几种神经发育障碍相关的风险基因和生物学途径，包括脆性 X 综合征、自闭症谱系障碍和 CHARGE 综合征。虽然这些既定的果蝇行为范式在历史上一直被用于研究遗传风险因素，但它们还有许多其他应用，包括在发育神经毒理学研究中用于确定神经发育障碍的环境风险因素。然而，缺乏逐步描述如何将学习和记忆测定应用于果蝇发育神经毒理学研究的协议。在这里，我们描述了两种用于果蝇的定量行为范式——捕食者诱导的产卵和求偶条件反射——可用于在发育神经毒理学研究中测量不同形式的学习和记忆。非联想学习和记忆在这里通过检查雌性果蝇对内生寄生蜂的产卵行为来测量，而在雄性中使用求偶条件反射来测量联想学习和记忆。我们的方案概述了对发育中的果蝇进行口服毒物暴露、内生寄生蜂培养、测量果蝇产卵活动以及评估条件求偶的程序，以确定毒物对雌性和雄性学习和记忆的影响。作为一个例子，我们使用双酚 A 展示了这些方案，双酚 A 是一种用于合成聚碳酸酯塑料的化学物质，以确定其对学习和记忆的影响。这些方案成本低廉，操作相对简单，对于刚开始进行果蝇行为研究的实验室，可以用来评估毒物暴露如何影响雄性和雌性果蝇的学习和记忆。
