以斑马鱼为模型系统的三室选择行为任务
Summary
我们提出了一个行为室，旨在评估认知性能。我们提供的数据表明，一旦获得，斑马鱼记得任务8周后。我们还表明，高血糖斑马鱼改变了认知能力，表明这种模式适用于评估认知和记忆的研究。
Abstract
神经退行性疾病是年龄依赖，衰弱和不治之症。最近的报告还把高血糖与记忆和/或认知障碍的变化联系起来。我们修改并开发了一个三室选择认知任务，类似于与啮齿动物一起用于高血糖斑马鱼的任务。测试室由位于中央的起始室和两侧的两个选择舱组成，并配有一个浅滩，用作奖励。我们提供的数据表明，一旦获得，斑马鱼记得任务至少8周后。我们的数据表明，斑马鱼对这一奖励反应强烈，经过4周的治疗，我们已经发现了高血糖鱼的认知缺陷。此行为分析可能也适用于与认知和记忆相关的其他研究。
Introduction
神经退行性疾病是年龄依赖，衰弱和不治之症。这些疾病的流行率正在上升，因此迫切需要改进和制定新的治疗战略。每种疾病的发病和表现是独一无二的，因为有些会影响语言、运动和自主大脑区域，而另一些疾病则会导致学习缺陷和记忆丧失。最值得注意的是，认知缺陷和/或损伤是所有神经退行性疾病中最普遍的并发症2。为了揭示这些神经退行性疾病所涉及的基本机制，采用了许多不同的模型系统（包括单细胞生物对 嗜好 动物，如啮齿动物和人类等高阶脊椎动物）的使用：然而，大多数神经退行性疾病仍然不治之症。
学习和记忆是生物体之间高度保守的过程，因为环境的不断变化需要适应3。认知和突触可塑性的损伤已经在几个啮齿动物模型中得到了证明。具体来说，成熟的行为分析使用关联学习来评估各种损伤引起的疾病和紊乱后的认知变化。此外，对比歧视逆转评估认知缺陷，因为它涉及到更高的顺序学习和记忆功能，而反转取决于对以前学会的关联的抑制。广泛使用的三室选择任务阐明了中枢神经系统5，6的学习和记忆通路可能的缺陷。最近，这个领域已经扩大到包括非哺乳动物模型，如斑马鱼（Danio rerio），因为几个范式已经开发了从幼虫到成人7，8的年龄范围。
斑马鱼提供了复杂性和简单性的平衡，有利于用行为技术评估认知障碍。首先，斑马鱼由于体积小、繁殖性大，可接受高通量的行为筛查。其次，斑马鱼有一个结构，横向苍白，这是类似于哺乳动物海马，因为它有类似的神经元标记和细胞类型7。斑马鱼也能够获取和记住空间信息9，并且，像人类一样，是日间10。因此，斑马鱼被用作神经退行性疾病的模型，频率越来越高，这并不奇怪。然而，由于缺乏适当的行为检测，因此很难将斑马鱼模型应用于认知评估。使用斑马鱼特有的行为分析出版的工作包括关联学习任务11，焦虑行为12，记忆13，对象识别14，和条件地点偏好15，16，17，18，19。虽然斑马鱼的行为检测已经取得了许多进展，但啮齿动物认知功能的一些测试的对应物尚未开发用于斑马鱼18。
基于我们实验室先前的研究，我们根据使用社交互动作为奖励与啮齿动物使用的三室选择任务，对斑马鱼的认知任务进行了建模/开发。此外，我们扩展了行为任务的关联学习方面，并纳入了对比度歧视逆转，希望进一步发展这一行为任务，以评估认知障碍。这使我们能够审查歧视学习的最初获得以及随后在逆转阶段对这种学习的抑制。在目前的研究中，我们证明这个程序提供了一个可靠的方法来评估斑马鱼在葡萄糖浸泡后4或8周的认知功能。
Protocol
Representative Results
Discussion
虽然在过去15年中，使用斑马鱼进行的神经科学研究的数量和种类都出现了巨大的增长，但与哺乳动物模型系统11、25、26相比，该物种缺乏行为检测。在这里，我们展示了一个三室选择任务，开发用于啮齿动物可以调整，以评估在斑马鱼视觉歧视学习的获取和逆转。这项任务以活浅滩作为奖励，提供了一个强?…
Declarações
The authors have nothing to disclose.
Acknowledgements
我们感谢萨布丽娜·琼斯协助她使啮齿动物三室选择模式适应斑马鱼模型，杰里米·波波维茨和艾莉森·默克在行为收集日、跑步试验、动物护理和水箱设置方面的帮助。特别感谢詹姆斯·福布斯（机械工程师）协助三室选择坦克的设计和施工。
资助：VPC和TLD获得了美国大学艺术与科学学院的联合教师研究支持补助金（FRSG）。CJR 得到了美国大学艺术与科学学院研究生支持的支持。
Materials
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| Marineland® Silicone Aquarium Sealant | Petsmart | Item #2431002 | |
| PVC (Polyvinyl Chloride) Sheet, Opaque Gray, Standard Tolerance, UL 94/ASTM D1784, 0.125" Thickness, 12" Width, 24" Length | Amazon | N/A | https://www.amazon.com/Polyvinyl-Chloride-Standard-Tolerance-Thickness/dp/B000MAMGEQ/ref=sr_1_2?dchild=1&keywords=grey+PVC+sheet&qid=1597081440&sr=8-2 |
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| Tetra 10 Gallon Fish tank | Petsmart | Item #5271256 | |
| Top Fin Fine Mesh Fish Net (3 in) | Petsmart | Item #5175115 |
Referências
- Gitler, A. D., Dhillon, P., Shorter, J. Neurodegenerative disease: models, mechanisms, and a new hope. Disease Models & Mechanisms. 10, 499-502 (2017).
- Perry, R. J., Watson, P., Hodges, J. R. The nature and staging of attention dysfunction in early (minimal and mild) Alzheimer’s disease: relationship to episodic and semantic memory impairment. Neuropsychologia. 38, 252-271 (2000).
- Gerlai, R. Learning and memory in zebrafish (Danio rerio). Methods in Cell Biology. 134, (2016).
- Davidson, T. L., et al. The effects of a high-energy diet on hippocampal-dependent discrimination performance and blood-brain barrier integrity differ for diet-induced obese and diet-resistant rats. Physiology and Behavior. 107, 26-33 (2012).
- Yang, M., Silverman, J. L., Crawley, J. N. Automated three-chambered social approach task for mice. Current Protocols in Neuroscience. 56 (1), (2011).
- Remmelink, E., Smit, A. B., Verhage, M., Loos, M. Measuring discrimination- and reversal learning in mouse models within 4 days and without prior food deprivation. Learning and Memory. 23, 660-667 (2016).
- Salas, C., et al. Neuropsychology of learning and memory in teleost fish. Zebrafish. 3, 157-171 (2006).
- Kalueff, A. V., et al. Towards a comprehensive catalog of zebrafish behavior 1.0 and beyond. Zebrafish. 10, 70-86 (2013).
- Luchiaria, A. C., Salajanb, D. C., Gerlai, R. Acute and chronic alcohol administration: Effects on performance of zebrafish in a latent learning task. Behavior Brain Research. 282, 76-83 (2015).
- Fadool, J., Dowling, J. Zebrafish: A model system for the study of eye genetics. Progress in Retinal and Eye Research. 27, 89-110 (2008).
- Fernandes, Y. M., Rampersad, M., Luchiari, A. C., Gerlai, R. Associative learning in the multichamber tank: A new learning paradigm for zebrafish. Behavioural Brain Research. 312, 279-284 (2016).
- Reider, M., Connaughton, V. P. Developmental exposure to methimazole increases anxiety behavior in zebrafish. Behavioral Neuroscience. , (2015).
- Capiotti, K. M., et al. Hyperglycemia induces memory impairment linked to increased acetylcholinesterase activity in zebrafish (Danio rerio). Behavioural Brain Research. 274, 319-325 (2014).
- May, Z., et al. Object recognition memory in zebrafish. Behavioural Brain Research. 296, 199-210 (2016).
- Mathur, P., Lau, B., Guo, S. Conditioned place preference behavior in zebrafish. Nature Protocols. 6, 338-345 (2011).
- Guo, S. Linking genes to brain, behavior and neurological diseases: What can we learn from zebrafish. Genes, Brain and Behavior. 3, 63-74 (2004).
- Kily, L. J. M., et al. Gene expression changes in a zebrafish model of drug dependency suggest conservation of neuro-adaptation pathways. Journal of Experimental Biology. 211, 1623-1634 (2008).
- Webb, K. J., et al. Zebrafish reward mutants reveal novel transcripts mediating the behavioral effects of amphetamine. Genome Biology. 10, (2009).
- Clayman, C. L., Malloy, E. J., Kearns, D. N., Connaughton, V. P. Differential behavioral effects of ethanol pre-exposure in male and female zebrafish (Danio rerio). Behavioural Brain Research. 335, 174-184 (2017).
- Ruhl, T., et al. Acute administration of THC impairs spatial but not associative memory function in zebrafish. Psychopharmacology. 231, 3829-3842 (2014).
- Gellermann, L. W. Chance orders of alternating stimuli in visual discrimination experiments. The Pedagogical Seminary and Journal of Genetic Psychology. 42, 206-208 (1933).
- Gleeson, M., Connaughton, V., Arneson, L. S. Induction of hyperglycaemia in zebrafish (Danio rerio) leads to morphological changes in the retina. Acta Diabetologica. 44, 157-163 (2007).
- Connaughton, V. P., Baker, C., Fonde, L., Gerardi, E., Slack, C. Alternate immersion in an external glucose solution differentially affects blood sugar values in older versus younger zebrafish adults. Zebrafish. 13, 87-94 (2016).
- Goldsmith, J. R., Jobin, C. Think small: Zebrafish as a model system of human pathology. Journal of Biomedicine and Biotechnology. 2012, 817341 (2012).
- Kalueff, A. V., Stewart, A. M., Gerlai, R., Court, P. Zebrafish as an emerging model for studying complex brain disorders. Trends in Pharmacological Sciences. 35, 63-75 (2014).
- Gerlai, R. Associative learning in zebrafish (Danio rerio). Methods in cell biology. 101, 249-270 (2011).
