Zebrafish最新文献_第4页

Optimization of Zebrafish Larvae 6-OHDA Exposure for Neurotoxin Induced Dopaminergic Marker Reduction. 优化斑马鱼幼体的 6-OHDA 暴露，以减少神经毒素诱导的多巴胺能标记物。

Zebrafish Pub Date : 2024-04-12 DOI: 10.1089/zeb.2023.0098

Adrian Romero, Armando Sanchez, Jocelyn D Jones, Kristel Ledesma, M. El-Halawany, Ayman K Hamouda, Brent R Bill

{"title":"Optimization of Zebrafish Larvae 6-OHDA Exposure for Neurotoxin Induced Dopaminergic Marker Reduction.","authors":"Adrian Romero, Armando Sanchez, Jocelyn D Jones, Kristel Ledesma, M. El-Halawany, Ayman K Hamouda, Brent R Bill","doi":"10.1089/zeb.2023.0098","DOIUrl":"https://doi.org/10.1089/zeb.2023.0098","url":null,"abstract":"Parkinson's disease (PD) is a neurodegenerative disorder that is clinically assessed by motor symptoms associated with the loss of midbrain dopaminergic neurons affecting the quality of life for over 8.5 million people worldwide. The neurotoxin 6-hydroxydopamine (6-OHDA) has been used to chemically induce a PD-like state in zebrafish larvae by several laboratories; however, highly variable concentration, methodology, and reagents have resulted in conflicting results suggesting a need to investigate these issues of reproducibility. We propose a protocol that addresses the differences in methodology and induces changes in 6 days postfertilization (dpf) larvae utilizing a 24-h exposure at 3 dpf with 30 μM 6-OHDA. Despite ∼50% lethality, no morphological or development differences in surviving fish are observed. Definition of our model is defined by downregulation of the expression of th1 by reverse transcriptase-quantitative polymerase chain reaction, a marker for dopaminergic neurons and a reduction in movement. Additionally, we observed a downregulation of pink1 and an upregulation of sod1 and sod2, indicators of mitochondrial dysfunction and response to reactive oxygen species, respectively.","PeriodicalId":94273,"journal":{"name":"Zebrafish","volume":"9 6","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-04-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140712071","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Zebrafish as Versatile Model for Assessing Animal Venoms and Toxins: Current Applications and Future Prospects. 斑马鱼作为评估动物毒液和毒素的多功能模型：当前应用与未来展望

Zebrafish Pub Date : 2024-04-12 DOI: 10.1089/zeb.2023.0088

Fajar Sofyantoro, Nur Indah Septriani, D. S. Yudha, Ega Adhi Wicaksono, D. Priyono, Wahyu Aristyaning Putri, Alfian Primahesa, Anita Restu Puji Raharjeng, Y. A. Purwestri, T. R. Nuringtyas

{"title":"Zebrafish as Versatile Model for Assessing Animal Venoms and Toxins: Current Applications and Future Prospects.","authors":"Fajar Sofyantoro, Nur Indah Septriani, D. S. Yudha, Ega Adhi Wicaksono, D. Priyono, Wahyu Aristyaning Putri, Alfian Primahesa, Anita Restu Puji Raharjeng, Y. A. Purwestri, T. R. Nuringtyas","doi":"10.1089/zeb.2023.0088","DOIUrl":"https://doi.org/10.1089/zeb.2023.0088","url":null,"abstract":"Animal venoms and toxins hold promise as sources of novel drug candidates, therapeutic agents, and biomolecules. To fully harness their potential, it is crucial to develop reliable testing methods that provide a comprehensive understanding of their effects and mechanisms of action. However, traditional rodent assays encounter difficulties in mimicking venom-induced effects in human due to the impractical venom dosage levels. The search for reliable testing methods has led to the emergence of zebrafish (Danio rerio) as a versatile model organism for evaluating animal venoms and toxins. Zebrafish possess genetic similarities to humans, rapid development, transparency, and amenability to high-throughput assays, making it ideal for assessing the effects of animal venoms and toxins. This review highlights unique attributes of zebrafish and explores their applications in studying venom- and toxin-induced effects from various species, including snakes, jellyfish, cuttlefish, anemones, spiders, and cone snails. Through zebrafish-based research, intricate physiological responses, developmental alterations, and potential therapeutic interventions induced by venoms are revealed. Novel techniques such as CRISPR/Cas9 gene editing, optogenetics, and high-throughput screening hold great promise for advancing venom research. As zebrafish-based insights converge with findings from other models, the comprehensive understanding of venom-induced effects continues to expand, guiding the development of targeted interventions and promoting both scientific knowledge and practical applications.","PeriodicalId":94273,"journal":{"name":"Zebrafish","volume":"126 2","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-04-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140708841","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

An Undergraduate Course in CRISPR/Cas9-Mediated Gene Editing in Zebrafish. 斑马鱼 CRISPR/Cas9 基因编辑本科课程。

Zebrafish Pub Date : 2024-04-01 DOI: 10.1089/zeb.2023.0091

Renu Srivastava, Connor W Davison, Abigail G Krull, Seth M Entriken, Amanda Zumbrock, Maria Daniela Cortes Hidalgo, Kiernan J Adair, Anna M Escherich, Jonathan N Lara, Emma C Neverman, Megan Hodnefield, Elyse McElligtot, Elizabeth J Sandquist, Craig Ogilvie, Pascal J Lafontant, J. Essner

{"title":"An Undergraduate Course in CRISPR/Cas9-Mediated Gene Editing in Zebrafish.","authors":"Renu Srivastava, Connor W Davison, Abigail G Krull, Seth M Entriken, Amanda Zumbrock, Maria Daniela Cortes Hidalgo, Kiernan J Adair, Anna M Escherich, Jonathan N Lara, Emma C Neverman, Megan Hodnefield, Elyse McElligtot, Elizabeth J Sandquist, Craig Ogilvie, Pascal J Lafontant, J. Essner","doi":"10.1089/zeb.2023.0091","DOIUrl":"https://doi.org/10.1089/zeb.2023.0091","url":null,"abstract":"We have developed a one-credit semester-long research experience for undergraduate students that involves the use of CRISPR/Cas9 to edit genes in zebrafish. The course is available to students at all stages of their undergraduate training and can be taken up to four times. Students select a gene of interest to edit as the basis of their semester-long project. To select a gene, exploration of developmental processes and human disease is encouraged. As part of the course, students use basic bioinformatic tools, design guide RNAs, inject zebrafish embryos, and analyze both the molecular consequences of gene editing and phenotypic outcomes. Over the 10 years we have offered the course, enrollment has grown from less than 10 students to more than 60 students per semester. Each year, we choose a different gene editing strategy to explore based on recent publications of gene editing methodologies. These have included making CRISPants, targeted integrations, and large gene deletions. In this study, we present how we structure the course and our assessment of the course over the past 3 years.","PeriodicalId":94273,"journal":{"name":"Zebrafish","volume":"17 6","pages":"162-170"},"PeriodicalIF":0.0,"publicationDate":"2024-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140763587","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Bringing Real Inquiry-Based Science to Diverse Secondary Educational Environments: A Virtual Zebrafish Laboratory to Investigate Environmental Health. 将真正的探究式科学引入多样化的中学教育环境：研究环境健康的虚拟斑马鱼实验室。

Zebrafish Pub Date : 2024-04-01 DOI: 10.1089/zeb.2023.0087

M. Carvan, Thomas Hansen, Renee A. Hesselbach, Amy Zientek, Craig A. Berg, David H. Petering

{"title":"Bringing Real Inquiry-Based Science to Diverse Secondary Educational Environments: A Virtual Zebrafish Laboratory to Investigate Environmental Health.","authors":"M. Carvan, Thomas Hansen, Renee A. Hesselbach, Amy Zientek, Craig A. Berg, David H. Petering","doi":"10.1089/zeb.2023.0087","DOIUrl":"https://doi.org/10.1089/zeb.2023.0087","url":null,"abstract":"The goal of the University of Wisconsin-Milwaukee WInSTEP SEPA program is to provide valuable and relevant research experiences to students and instructors in diverse secondary educational settings. Introducing an online experience allows the expansion of a proven instructional research program to a national scale and removes many common barriers. These can include lack of access to zebrafish embryos, laboratory equipment, and modern classroom facilities, which often deny disadvantaged and underrepresented students from urban and rural school districts valuable inquiry-based learning opportunities. An online repository of zebrafish embryo imagery was developed in the Carvan laboratory to assess the effects of environmental chemicals. The WInSTEP SEPA program expanded its use as an accessible online tool, complementing the existing classroom experience of our zebrafish module. This virtual laboratory environment contains images of zebrafish embryos grown in the presence of environmental toxicants (ethanol, caffeine, and nicotine), allowing students to collect data on 19 anatomical endpoints and generate significant amounts of data related to developmental toxicology and environmental health. This virtual laboratory offers students and instructors the choice of data sets that differ in the independent variables of chemical concentration and duration of postfertilization exposure. This enables students considerable flexibility in establishing their own experimental design to match the curriculum needs of each instructor.","PeriodicalId":94273,"journal":{"name":"Zebrafish","volume":"90 ","pages":"73-79"},"PeriodicalIF":0.0,"publicationDate":"2024-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140794206","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Tagging the tjp1a Gene in Zebrafish with Monomeric Red Fluorescent Protein Using Biotin Homology Arms. 用生物素同源臂为斑马鱼的 tjp1a 基因标记单体红色荧光蛋白。

Zebrafish Pub Date : 2024-04-01 DOI: 10.1089/zeb.2023.0096

Connor W Davison, Hamelynn Harzman, Jessie Nicholson, Seth M Entriken, Kierinn Mobley, Abigail G Krull, Manik Singhal, Caleb Skow, Nathan Matthews, Lindsey Kopp, Benjamin Gillette, Tyler J Weide, Jana R Hukvari, Sofia C P Stumpf, Olivia M Feldmann, M. McGrail, Renu Srivastava, J. Essner

{"title":"Tagging the tjp1a Gene in Zebrafish with Monomeric Red Fluorescent Protein Using Biotin Homology Arms.","authors":"Connor W Davison, Hamelynn Harzman, Jessie Nicholson, Seth M Entriken, Kierinn Mobley, Abigail G Krull, Manik Singhal, Caleb Skow, Nathan Matthews, Lindsey Kopp, Benjamin Gillette, Tyler J Weide, Jana R Hukvari, Sofia C P Stumpf, Olivia M Feldmann, M. McGrail, Renu Srivastava, J. Essner","doi":"10.1089/zeb.2023.0096","DOIUrl":"https://doi.org/10.1089/zeb.2023.0096","url":null,"abstract":"Tjp1a and other tight junction and adherens proteins play important roles in cell-cell adhesion, scaffolding, and forming seals between cells in epithelial and endothelial tissues. In this study, we labeled Tjp1a of zebrafish with the monomeric red fluorescent protein (mRFP) using CRISPR/Cas9-mediated targeted integration of biotin-labeled polymerase chain reaction (PCR) generated templates. Labeling Tjp1a with RFP allowed us to follow membrane and junctional dynamics of epithelial and endothelial cells throughout zebrafish embryo development. For targeted integration, we used short 35 bp homology arms on each side of the Cas9 genomic target site at the C-terminal of the coding sequence in tjp1a. Through PCR using 5' biotinylated primers containing the homology arms, we generated a double-stranded template for homology directed repair containing a flexible linker followed by RFP. Cas9 protein was complexed with the tjp1a gRNA before mixing with the repair template and microinjected into one-cell zebrafish embryos. We confirmed and recovered a precise integration allele at the desired site at the tjp1a C-terminus. Examination of fluorescence reveals RFP cell-cell junctional labeling using confocal imaging. We are currently using this stable tjp1a-mRFPis86 line to examine the behavior and interactions between cells during vascular formation in zebrafish.","PeriodicalId":94273,"journal":{"name":"Zebrafish","volume":"640 ","pages":"191-197"},"PeriodicalIF":0.0,"publicationDate":"2024-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140791119","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

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

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

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

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

{"title":"Development of an Undergraduate Cell Biology Laboratory to Assess Pigmentation and Cell Size in a Zebrafish Model of Uveal Melanoma.","authors":"Andrea M Henle","doi":"10.1089/zeb.2023.0095","DOIUrl":"10.1089/zeb.2023.0095","url":null,"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.","PeriodicalId":94273,"journal":{"name":"Zebrafish","volume":"21 2","pages":"137-143"},"PeriodicalIF":0.0,"publicationDate":"2024-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140873294","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0