Modeling Fetal Alcohol Spectrum Disorders in Zebrafish to Characterize the Impact of an Adverse Embryonic Environment on Adult Social Behavior

Mindy Rampersad; Yohaan Fernandes

doi:10.3791/65834

JoVE Journal
Behavior

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JoVE Journal Behavior

Modeling Fetal Alcohol Spectrum Disorders in Zebrafish to Characterize the Impact of an Adverse Embryonic Environment on Adult Social Behavior

对斑马鱼的胎儿酒精谱系障碍进行建模，以表征不良胚胎环境对成人社交行为的影响

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04:58 min

February 9, 2024

DOI: 10.3791/65834-v

Mindy Rampersad¹, Yohaan Fernandes¹

¹Department of Biology,University of South Dakota

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Overview

This protocol outlines the steps to establish a social preference assay for adult zebrafish, demonstrating its use in characterizing ethanol-induced social defects.

Key Study Components

Area of Science

Neuroscience
Behavioral Biology
Genetics

Background

Zebrafish are used to model the effects of environmental factors on behavior.
Alcohol exposure during development can lead to changes in gene expression and behavior.
Fetal alcohol spectrum disorders can result in social deficits, which this study aims to model.
Automated tracking and CRISPR technology are utilized to enhance research capabilities.

Purpose of Study

To establish a social preference assay for adult zebrafish.
To investigate the social deficits associated with embryonic alcohol exposure.
To determine the impact of low-dose alcohol on social behavior in zebrafish.

Methods Used

Setup of 1.4-liter and 37-liter tanks for behavioral assays.
Use of white corrugated plastic to enhance visual contrast.
Randomization of trials using an online generator.
Observation of social interactions and shoaling behavior in zebrafish.

Main Results

Control fish showed a stronger shoaling response compared to ethanol-treated fish.
Ethanol exposure blunted the shoaling response in zebrafish.
No difference in mobility was observed between groups during habituation.
Experimental fish spent less time near social stimuli when exposed to alcohol.

Conclusions

Zebrafish can effectively model social deficits related to fetal alcohol exposure.
Low-dose alcohol exposure during development affects social behavior.
This assay can be used to further investigate gene-environment interactions.

Frequently Asked Questions

What is the significance of using zebrafish in this study?

Zebrafish are a valuable model for studying the effects of environmental factors on behavior due to their genetic similarity to humans and transparent embryos.

How does alcohol exposure affect zebrafish behavior?

Alcohol exposure during development can lead to social deficits, which are modeled in this study using a social preference assay.

What methods are used to track zebrafish behavior?

Automated tracking systems are employed to increase throughput and accuracy in observing zebrafish behavior during assays.

What are fetal alcohol spectrum disorders?

Fetal alcohol spectrum disorders are a range of effects that can occur in an individual whose mother drank alcohol during pregnancy, including social and behavioral deficits.

What is the purpose of the social preference assay?

The assay is designed to assess social behavior in zebrafish and to identify the impact of environmental factors like alcohol exposure on these behaviors.

How does this research contribute to understanding genetic profiles?

This research aims to explore how genetic profiles may influence susceptibility to the effects of embryonic alcohol exposure on behavior.

当前方案的目标是概述建立和使用成年斑马鱼社会偏好测定的必要步骤，并证明它可用于表征乙醇诱导的社会缺陷。

我们实验室的范围是了解基因如何与大脑发育相互作用以影响行为。在我们的实验室中，我们使用斑马鱼来了解发育过程中酒精暴露等环境因素如何导致斑马鱼基因发生变化，从而影响大脑发育并导致行为改变。我们的总体目标是了解一个人的遗传特征是否使他们或多或少可能受到胚胎酒精暴露的影响。因此，在我们的领域，我们目前正在使用自动跟踪，这可以提高吞吐量，并且我们正在使用 CRISPR 技术等先进的基因编辑。将这两者配对在一起使我们能够进一步破译基因-环境相互作用如何导致行为变化。因此，我们第一次能够证明斑马鱼可用于模拟与胎儿酒精谱系障碍相关的社会缺陷。过去，斑马鱼被用来模拟产前酒精暴露的更严重结果。具体来说，它们的用途是寻找因长时间服用高酒精剂量而引起的物理缺陷。作为一个领域，我们不知道斑马鱼是否可以用来模拟与胎儿酒精谱系障碍相关的社会缺陷。我们首次能够证明，在很短的时间内使用低剂量的酒精会导致成年斑马鱼出现社会缺陷。

[导师]首先，将 1.4 升油箱沿着 37 升油箱的宽度放置。用白色波纹塑料覆盖37升水箱的后部和底部，以增强实验鱼与背景的对比度。然后将波纹塑料放置在1.4升水箱的外壁上，以增强对实验鱼的社会刺激对比度。接下来，在 1.4 升和 37 升的水箱之间插入白色波纹塑料，以防止鱼在栖息期间看到对方。将相机放置在适当的距离，以捕捉 37 升油箱的长度和 1.4 升油箱的一半。使用配备 15 瓦 T8 全光谱灯的水族箱罩灯照亮 37 升水箱。在进行行为测定之前，使用在线随机序列生成器对所有试验进行随机化。然后用与外壳架相匹配的水填充指定用于行为测定的 37 升水箱。确保水温与外壳架在 2 摄氏度范围内一致，并且所有水箱中的水位相同。沿着 37 升水箱的顶部和底部长度标记 5 厘米的增量，为 50 厘米的测试罐创建 10 个区域，每个区域跨度为 50 厘米。接下来，从同一队列中选择两只雄性和两只雌性作为实验鱼作为社会刺激。用网在养箱中捕获 16 周大的实验鱼，并将其放入装有鱼水的容器中。然后将实验鱼放置在测试场的中心。在开始 20 分钟试用之前，请检查检测设置是否符合所选软件的用户偏好。在最初的 10 分钟内，保持 37 升和 1.4 升油箱之间的不透明屏障。然后，小心地去除不透明的障碍，让实验鱼看到社会刺激。胚胎乙醇暴露减弱了浅滩反应，因为这些鱼无法靠近浅滩。与暴露于 1% 酒精的鱼相比，对照鱼对活浅滩的反应明显更强。在存在社会刺激的情况下，与乙醇处理的鱼相比，对照鱼在 1 区停留的时间更多。胚胎乙醇暴露不会损害活动能力，因为在没有社会刺激的情况下，在习惯化过程中，跨区域的组之间没有差异。当社会刺激不可见时，鱼在 1 区度过的时间没有差异。

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