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基于图像的小分子筛选以鉴定斑马鱼多巴胺能神经元神经保护化合物的方案。

Protocol for image-based small-molecule screen to identify neuroprotective compounds for dopaminergic neurons in zebrafish.

机构信息

Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, San Francisco, CA, USA.

Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, San Francisco, CA, USA.

出版信息

STAR Protoc. 2024 Mar 15;5(1):102837. doi: 10.1016/j.xpro.2024.102837. Epub 2024 Jan 12.

DOI:10.1016/j.xpro.2024.102837
PMID:38217853
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10825766/
Abstract

Whole-organism-based screen holds promise for discovering biologically active compounds. However, high-content imaging is challenging due to the difficulty of positioning live animals and individual variability of neuron counts. Here, we present a protocol to identify neuroprotective compounds for dopaminergic neurons in zebrafish using an image-based small-molecule screen. We describe steps for raising larvae, agarose embedding, and treatment to induce neurodegeneration. We then detail procedures for live confocal imaging, image processing, and data analysis. For complete details on the use and execution of this protocol, please refer to Kim et al. (2021)..

摘要

基于整体生物的筛选有望发现具有生物活性的化合物。然而，由于活体动物定位和神经元计数个体差异的困难，高内涵成像具有挑战性。在这里，我们提出了一种使用基于图像的小分子筛选来鉴定斑马鱼中多巴胺能神经元的神经保护化合物的方案。我们描述了饲养幼虫、琼脂糖包埋和诱导神经退行性变的处理步骤。然后，我们详细介绍了活共聚焦成像、图像处理和数据分析的程序。有关此方案使用和执行的完整详细信息，请参阅 Kim 等人。(2021)。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e1a1/10825766/f3d11ffb748e/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e1a1/10825766/884d8cc1a8d1/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e1a1/10825766/2aedc168f51e/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e1a1/10825766/b2bee46e5a3a/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e1a1/10825766/8b7ad3102ae3/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e1a1/10825766/0b88aed4ec09/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e1a1/10825766/eb4ff2b9fe1c/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e1a1/10825766/f3d11ffb748e/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e1a1/10825766/884d8cc1a8d1/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e1a1/10825766/2aedc168f51e/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e1a1/10825766/b2bee46e5a3a/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e1a1/10825766/8b7ad3102ae3/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e1a1/10825766/0b88aed4ec09/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e1a1/10825766/eb4ff2b9fe1c/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e1a1/10825766/f3d11ffb748e/gr6.jpg

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本文引用的文献

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Front Pharmacol. 2022 Mar 18;13:837756. doi: 10.3389/fphar.2022.837756. eCollection 2022.
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Elife. 2021 Sep 22;10:e69795. doi: 10.7554/eLife.69795.
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