Zebrafish最新文献

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Fipronil Affects Craniofacial and Heart Development in Zebrafish Embryos (Danio rerio). 氟虫腈对斑马鱼胚胎（Danio rerio）颅面和心脏发育的影响

Zebrafish

Pub Date : 2024-04-01 DOI: 10.1089/zeb.2023.0055

Kasey L Cooper, Zoe G Krut, Bennet D Franz, Benjamin S Walker, Alexander G Kramer, Jonathan R Morgan, Christopher S. Lassiter

Fipronil is a broad-spectrum insecticide that has off-target effects in developing vertebrate embryos. In this study, we investigate treatment of zebrafish embryos with fipronil over the course of 5 days and examine the effects on body length, the cardiovascular system, and craniofacial morphology. We found the insecticide caused shorter body length and a decrease in eye size. By examining specific heart chamber morphology, as well as jaw angle and length, we quantified defects including enlargement of the heart and increases in jaw length and width. Further studies are needed to assess the mechanisms of fipronil's effect on vertebrate development for both environmental and human health concerns.

氟虫腈是一种广谱杀虫剂，对发育中的脊椎动物胚胎有脱靶效应。在本研究中，我们调查了用氟虫腈处理斑马鱼胚胎 5 天的过程，并研究了其对体长、心血管系统和颅面形态的影响。我们发现，杀虫剂会导致斑马鱼体长缩短、眼睛变小。通过检查具体的心腔形态以及下颌角度和长度，我们量化了包括心脏增大、下颌长度和宽度增加在内的缺陷。我们需要进一步研究氟虫腈对脊椎动物发育的影响机制，以解决环境和人类健康问题。

引用次数: 0

Real-Time Observation of Germinal Vesicle Migration During Oocyte Meiotic Cell Division Using Ovarian Fluorescent Transgenic Zebrafish. 利用卵巢荧光转基因斑马鱼实时观察卵母细胞减数分裂过程中胚芽囊泡的迁移。

Zebrafish

Pub Date : 2024-04-01 DOI: 10.1089/zeb.2023.0048

Eisei Tsutsumi, Toshinobu Tokumoto

The transgenic (TG) zebrafish allows researchers to bio-image specific biological phenomena in cells and tissues in vivo. We established TG lines to monitor changes in the ovaries of live fish. The original TG line with ovarian fluorescence was occasionally established. Although the cDNA integrated into the line was constructed for the expression of enhanced green fluorescent protein (EGFP) driven by the medaka β-actin promoter, the expression of EGFP is restricted to the oocytes and gills in adult fish. Furthermore, we found that germinal vesicles (GVs) in oocytes of the established line can be observed by relatively strong fluorescence around the GV. In this study, we tried to capture the dynamic processes of germinal vesicle breakdown (GVBD) during meiotic cell division using the GV fluorescent oocytes. As a result, GV migration and GVBD could be monitored in real time. We also succeeded in observing actin filaments involved in the migration of GV to the animal pole. This strain can be used for education in the process of oocyte meiotic cell division.

通过转基因（TG）斑马鱼，研究人员可以对体内细胞和组织中的特定生物现象进行生物成像。我们建立 TG 品系是为了监测活鱼卵巢的变化。最初的 TG 品系偶尔会出现卵巢荧光。虽然该品系中整合的 cDNA 是由青鳉 β-肌动蛋白启动子驱动的增强型绿色荧光蛋白（EGFP）的表达，但 EGFP 的表达仅限于成鱼的卵母细胞和鳃。此外，我们还发现，已建立品系的卵母细胞中的生殖泡（GV）可通过 GV 周围相对较强的荧光观察到。在这项研究中，我们试图利用 GV 荧光卵母细胞捕捉减数分裂细胞分裂过程中生殖泡破裂（GVBD）的动态过程。因此，GV迁移和GVBD可被实时监测。我们还成功地观察到了参与 GV 向动物极迁移的肌动蛋白丝。该菌株可用于卵母细胞减数分裂过程的教育。

引用次数: 0

Growing Project BioEYES: A Reflection on 20 Years of Developing and Replicating a K-12 Science Outreach Program. 发展项目 BioEYES：开发和复制 K-12 科学推广计划 20 年的反思。

Zebrafish

Pub Date : 2024-04-01 DOI: 10.1089/zeb.2023.0059

Jamie R Shuda, Valerie G. Butler, Theresa M Nelson, Jaqueline M Davidson, Auset M Taylor, Steven A Farber

Project BioEYES celebrated 20 years in K12 schools during the 2022-2023 school year. Using live zebrafish (Danio rerio) during week-long science experiments, sparks the interest of students and teachers from school districts, locally and globally. Over the past two decades, BioEYES has been replicated in different ways based on the interest and capacity of our partners. This article discusses several of the successful models, the common challenges, and how each BioEYES site has adopted guiding principles to help foster their success. The core principles of (a) reinforcing content that students are expected to learn in schools, while focusing on the students BECOMING scientists through hands-on experimentation and (b) establishing trust and buy-in from collaborating teachers and partners are what has led to BioEYES being sustained and replicated over the past two decades.

2022-2023 学年，生物眼睛项目在 K12 学校开展了 20 周年庆典活动。在为期一周的科学实验中使用活体斑马鱼（Danio rerio），激发了本地和全球各学区学生和教师的兴趣。在过去的二十年里，根据合作伙伴的兴趣和能力，我们以不同的方式复制了 "生物眼睛 "项目。本文将讨论其中几种成功的模式、共同面临的挑战，以及每个 "生物国际年 "项目点如何采用指导原则来帮助其取得成功。核心原则包括：（a）强化学生在学校应学习的内容，同时关注学生通过动手实验成为科学家；（b）建立合作教师和合作伙伴的信任和支持，正是这些原则使生物教育项目在过去二十年中得以持续开展和推广。

引用次数: 0

Zebrafish in Education: Tackling Big Problems with Little Fish. 教育中的斑马鱼：用小鱼解决大问题。

Zebrafish

Pub Date : 2024-04-01 DOI: 10.1089/zeb.2024.0141

M. Jackstadt, Lara Hutson, Jennifer O Liang, M. Pickart, Christopher Pierret, T. Franz-Odendaal

引用次数: 0

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

Zebrafish

Pub Date : 2024-04-01 DOI: 10.1089/zeb.2023.0095

Andrea M Henle

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:GNAQ^Q209L)]. The transgenic fish express a hyperactive Gα protein, GNAQ^Q209L, 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:GNAQ^Q209L) 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.

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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引用次数: 0

The InSciEdRS View: A User-Friendly and Accessible Microscope with Easy-to-Follow Companion Curricula. InSciEdRS View：用户界面友好、易于使用的显微镜，以及简单易学的配套课程。

Zebrafish

Pub Date : 2024-04-01 DOI: 10.1089/zeb.2023.0093

Soaleha Shams, Sidney Olson, Michael P. Ekker, Adam Salmi, Stephen C Ekker, Christopher Pierret

Microscopes are essential for research and education in science. Unlike computers and online learning tools, however, microscopes are not currently a fixed element in K-12 classrooms, due to steep cost, needless complexity, and often requiring a prohibitive level of staff training to effectively deploy. In a collaboration with Area 10 Labs, Integrated Science Education Outreach (InSciEd Out) developed a state-of-the-art alternative microscope, the InSciEdRS View, to reduce the financial barrier, prohibitive per-student cost, unnecessary complexity, and extensive staff training. Utilizing a 1080p camera and a lunchbox-style case, this Wi-Fi- and USB-connectable microscope comes with all necessary components for visualization of microscopic specimens (10 × -50 × magnification). While built to handle the rigors of classroom use, its imaging capability and battery-operation can make it flexible for a laboratory or fieldwork as well. We further highlight here K-12 curricula that we have developed using larval zebrafish to enable teachers, science outreach leaders, and parents to support active hands-on science observations. The InSciEdRS View microscope and the InSciEd Out curricula are readily scalable, translatable, and accessible for traditional and neurodiverse students and integrating these in various settings can be an efficient way to achieve better outcomes in science education.

显微镜对于科学研究和教育至关重要。然而，与计算机和在线学习工具不同，显微镜目前还不是 K-12 教室的固定设备，原因是成本高昂、不必要的复杂性，而且往往需要对员工进行高水平的培训才能有效使用。综合科学教育拓展项目（InSciEd Out）与 Area 10 实验室合作，开发了一种最先进的替代显微镜--InSciEdRS View，以减少经济障碍、高昂的学生人均成本、不必要的复杂性和大量的人员培训。这款可连接 Wi-Fi 和 USB 的显微镜采用 1080p 摄像机和便当盒式外壳，配备了显微镜标本可视化（10 × -50 × 放大倍率）所需的所有组件。虽然该显微镜专为应对严格的课堂使用而设计，但其成像能力和电池操作使其在实验室或野外工作中也能灵活使用。我们在此进一步强调我们利用斑马鱼幼虫开发的 K-12 课程，使教师、科学推广领导者和家长能够支持积极的动手科学观察。InSciEdRS View显微镜和InSciEd Out课程可随时扩展、转化，并适合传统学生和神经多样性学生使用，将其整合到各种环境中可以有效地实现更好的科学教育成果。

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引用次数: 0

Lectins As Effective Tools in the Study of the Biliary Network and the Parenchymal Architecture of the Zebrafish (Danio rerio) Liver. 作为研究斑马鱼（Danio rerio）肝脏胆道网络和实质结构的有效工具的凝集素

Zebrafish

Pub Date : 2024-04-01 DOI: 10.1089/zeb.2023.0073

Hayden Bhavsar, Zoe Robinson, Matthew Benson, Feven Getachew, J. González-Rosa, Pascal J Lafontant

Lectins are carbohydrate-binding proteins with specific affinity to glycoconjugates expressed in various tissues. Lectins are of substantial utility as research, histochemical, and diagnostic tools in mammalian systems. Reactivity of 12 commonly used plant-based lectins was studied in zebrafish liver. Four lectins, tomato lectin (TL), wheat germ agglutinin, concanavalin A, and Jacalin showed strong reactivity to hepatic parenchymal structures. Importantly, TL reacted to glycoconjugates within segments of the larval and adult intrahepatic biliary network, from canaliculi to bile ducts. We provide evidence that lectins can serve as important histochemical tools to investigate the structural and functional characteristics of the zebrafish liver.

连接蛋白是一种碳水化合物结合蛋白，对各种组织中表达的糖结合物具有特异性亲和力。在哺乳动物系统中，凝集素是一种非常有用的研究、组织化学和诊断工具。我们研究了 12 种常用植物凝集素在斑马鱼肝脏中的反应性。四种凝集素，即番茄凝集素（TL）、小麦胚芽凝集素、凝集素 A 和 Jacalin 对肝实质结构表现出强烈的反应性。重要的是，番茄凝集素能与幼虫和成虫肝内胆管网络（从胆管到胆管）中的糖结合物发生反应。我们提供的证据表明，凝集素可作为研究斑马鱼肝脏结构和功能特征的重要组织化学工具。

引用次数: 0

Course on Breeding, Handling, and Experimental Procedures of Zebrafish (Danio rerio): Training, Refinement, Paradigm Shift, and Dissemination of Knowledge from Brazil to the World. 斑马鱼（Danio rerio）的繁殖、处理和实验程序课程：培训、改进、范式转变以及从巴西向世界传播知识。

Zebrafish

Pub Date : 2024-04-01 DOI: 10.1089/zeb.2023.0065

Tony Silveira, Mateus Kütter, B. Nornberg, Marcela Meirelles, Dennis Silva, Andrea Idelette López, Andressa Lopes, Caroline Castro, Jenifer Moraes, Lucia Emanueli Schimith, Priscila de Leon, Luis Fernando Marins, Mariana Remião

Currently, in Brazil, all researchers involved in animal experimentation must undergo training in laboratory animal science to stay updated on biology, methodology, ethics, and legal considerations related to the use of animals. The training program presented in this study not only aims to fulfill a legal obligation but also intends to train students and professionals to effectively care for their biomodels. It seeks to help them understand the importance of this care, both for the welfare of the animals and for the results of their projects. In total, 58 participants were present at the event (pre-event and full-time course). These participants consisted students and professionals from 11 institutions and 5 different countries. These numbers demonstrate the successful attainment of the desired capillarity in the scientific community and the posterior dissemination of knowledge. Through this course, it was possible to train the participants and raise their awareness about the importance of applying scientific knowledge in their daily practices to maintain the animals, ensuring the welfare of the models and refining the research. Finally, the program presented in this study, as well as the strategies adopted, can serve as a model for other institutions aiming to achieve similar results.

目前，在巴西，所有参与动物实验的研究人员都必须接受实验动物科学方面的培训，以了解与使用动物相关的生物学、方法学、伦理学和法律方面的最新情况。本研究中介绍的培训计划不仅旨在履行法律义务，还旨在培训学生和专业人员如何有效地照顾他们的生物模型。该计划旨在帮助他们了解这种照顾对动物福利和项目成果的重要性。共有 58 人参加了此次活动（活动前和全日制课程）。这些参与者包括来自 11 个机构和 5 个不同国家的学生和专业人员。这些数字表明，成功实现了科学界所期望的毛细管效应和知识的后续传播。通过该课程，可以对参与者进行培训，提高他们对在日常实践中应用科学知识维护动物、确保模型福利和完善研究的重要性的认识。最后，本研究中介绍的计划以及所采取的策略，可以为其他旨在取得类似成果的机构提供范例。

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引用次数: 0

Laboratory Course Using Zebrafish to Uncover Changing Roles of Wnt Signaling in Early Vertebrate Development. 利用斑马鱼揭示 Wnt 信号在脊椎动物早期发育中不断变化的作用的实验课程。

Zebrafish

Pub Date : 2024-04-01 DOI: 10.1089/zeb.2023.0070

Manuel Fiegl, Robin A Kimmel

Coordinated signaling pathway activity directs early patterning to set up the vertebrate body plan. Perturbations in the timing or location of signal molecule expression impacts embryo morphology and organ formation. In this study, we present a laboratory course to use zebrafish for studying the role of Wnt signaling in specifying the early embryonic axes. Students are exposed to basic techniques in molecular and developmental biology, including embryo manipulation, fluorescence microscopy, image processing, and data analysis. Furthermore, this course incorporates student-designed experiments to stimulate independent inquiry and improve scientific learning, providing an experience resembling graduate-level laboratory research. Students appreciated following vertebrate development in real-time, and principles of embryogenesis were reinforced by observing the morphological changes that arise due to signaling alterations. Scientific and research skills were enhanced through practice in experimental design, interpretation, and presentation.

协调的信号通路活动指导早期模式化，以建立脊椎动物的身体计划。信号分子表达时间或位置的干扰会影响胚胎形态和器官的形成。在本研究中，我们介绍了利用斑马鱼研究 Wnt 信号在指定早期胚胎轴中的作用的实验课程。学生将接触到分子和发育生物学的基本技术，包括胚胎操作、荧光显微镜、图像处理和数据分析。此外，本课程还结合了学生设计的实验，以激发独立探究和提高科学学习能力，为学生提供类似研究生水平实验室研究的体验。学生们非常喜欢实时跟踪脊椎动物的发育过程，并通过观察因信号改变而产生的形态变化，巩固了胚胎发生的原理。通过实验设计、解释和演示方面的练习，科学和研究技能得到了提高。

引用次数: 0

A 3D-Printed and Freely Available Device to Measure the Zebrafish Optokinetic Response Before and After Injury. 用于测量斑马鱼受伤前后视运动反应的三维打印和免费提供的装置。

Zebrafish

Pub Date : 2024-04-01 DOI: 10.1089/zeb.2023.0071

Ashley Hermans, Sophia Tajnai, Allison Tieman, Sarah Young, Ashley Franklin, Mackenzie Horutz, Steven J Henle

Zebrafish eyes are anatomically similar to humans and have a higher percentage of cone photoreceptors more akin to humans than most rodent models, making them a beneficial model organism for studying vision. However, zebrafish are different in that they can regenerate their optic nerve after injury, which most other animals cannot. Vision in zebrafish and many other vertebrate animals, including humans, can be accessed using the optokinetic response (OKR), which is an innate eye movement that occurs when tracking an object. Because fish cannot use an eye chart, we utilize the OKR that is present in virtually all vertebrates to determine if a zebrafish has vision. To this end, we have developed an inexpensive OKR setup that uses 3D-printed and off-the-shelf parts. This setup has been designed and used by undergraduate researchers and is also scalable to a classroom laboratory setup. We demonstrate that this setup is fully functional for assessing the OKR, and we use it to illustrate the return of the OKR following optic nerve injury in adult zebrafish.

斑马鱼的眼睛在解剖学上与人类相似，与大多数啮齿类动物模型相比，其视锥光感受器的比例更高，更接近人类，因此是研究视觉的有益模型生物。然而，斑马鱼的不同之处在于，它们的视神经在受伤后可以再生，而大多数其他动物却不能。斑马鱼和包括人类在内的许多其他脊椎动物的视觉可以通过光动力反应（OKR）获得，这是一种在追踪物体时发生的先天性眼球运动。由于鱼类不能使用眼图，我们利用几乎所有脊椎动物都有的 OKR 来确定斑马鱼是否有视觉。为此，我们开发了一种廉价的 OKR 装置，使用 3D 打印和现成的部件。该装置由本科生研究人员设计和使用，也可扩展为教室实验室装置。我们展示了这种装置在评估 OKR 方面的全部功能，并用它来说明成年斑马鱼视神经损伤后 OKR 的恢复情况。

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