Development of an Undergraduate Cell Biology Laboratory to Assess Pigmentation and Cell Size in a Zebrafish Model of Uveal Melanoma.

Zebrafish Pub Date : 2024-04-01 DOI:10.1089/zeb.2023.0095
Andrea M Henle
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Abstract

This study outlines a 2-week laboratory module for an authentic cell biology undergraduate research experience that uses zebrafish (Danio rerio), a popular model organism for research. Previous research has indicated that course-based undergraduate research experiences such as this one increase student confidence, active learning, and retention. During this research experience, students investigate variations in pigmentation in the caudal fins of wild type (WT) and transgenic fish [Tg(mitfa:GNAQQ209L)]. The transgenic fish express a hyperactive Gα protein, GNAQQ209L, under the melanocyte-specific mitfa promoter, offering insights into uveal melanoma, a common eye cancer. Students specifically analyze the black pigmented cells, melanophores, within the caudal fin. We determined that the transgenic zebrafish have increased pigmentation in their caudal fins, but smaller melanophores. These results suggest there are more melanophores in the Tg(mitfa:GNAQQ209L) fish compared to the WT. Future undergraduate research could investigate these cellular differences. This research experience imparts microscopy and image analysis skills and instills the ability to grapple with large datasets, statistical tests, and data interpretation in alignment with biology education principles. Post-laboratory surveys reveal students attain confidence in the above skills and in handling animals, along with a deeper appreciation for model organism research and its relevance to cancer cell biology.

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开发本科生细胞生物学实验室,以评估斑马鱼葡萄膜黑色素瘤模型中的色素沉着和细胞大小。
本研究概述了一个为期两周的真实细胞生物学本科生研究体验实验模块,该模块使用斑马鱼(Danio rerio)这一流行的研究模式生物。以往的研究表明,像这样以课程为基础的本科生研究体验可以增强学生的自信心,提高学生的主动学习能力和保留率。在这次研究体验中,学生们将研究野生型(WT)和转基因鱼[Tg(mitfa:GNAQQ209L)]尾鳍色素沉着的变化。转基因鱼在黑色素细胞特异性 mitfa 启动子的作用下表达高活性 Gα 蛋白 GNAQQ209L,为研究常见的眼癌--葡萄膜黑色素瘤提供了线索。学生们特别分析了尾鳍内的黑色素细胞--黑素细胞。我们发现,转基因斑马鱼尾鳍中的色素沉着增加了,但黑色素细胞却变小了。这些结果表明,与 WT 相比,Tg(mitfa:GNAQQ209L)斑马鱼体内有更多的黑色素细胞。未来的本科生研究可以调查这些细胞差异。这一研究经历传授了显微镜和图像分析技能,并培养了学生处理大型数据集、统计测试和数据解释的能力,使其与生物教育原则保持一致。实验后的调查显示,学生对上述技能和处理动物有了信心,同时对模式生物研究及其与癌细胞生物学的相关性有了更深刻的认识。
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