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Establishment of an In Vitro Assay for Analysis of the Mechanism Underlying Chorion Elevation in Zebrafish. 斑马鱼绒毛膜升高机制体外分析方法的建立。
IF 1.2
Zebrafish Pub Date : 2025-12-01 DOI: 10.1177/15458547251401451
Eisei Tsutsumi, Toshinobu Tokumoto
{"title":"Establishment of an <i>In Vitro</i> Assay for Analysis of the Mechanism Underlying Chorion Elevation in Zebrafish.","authors":"Eisei Tsutsumi, Toshinobu Tokumoto","doi":"10.1177/15458547251401451","DOIUrl":"https://doi.org/10.1177/15458547251401451","url":null,"abstract":"<p><p>Chorions, also known as egg membranes, form quickly after fertilization and play a role in subsequent embryonic development in fish. While the enzymatic hardening of the chorion due to the release of cortical alveoli (CA) components by the exocytosis of CA has been well-demonstrated, the initiation mechanism of this process has remained unresolved. Knockout lines with the <i>prss59.1</i> trypsin paralog gene exhibited abnormalities in chorion elevation. <i>Prss59.1</i> has been shown to be expressed on the chorion's surface. Therefore, we hypothesized that a trypsin-like enzyme expressed on the chorion could trigger chorion elevation. In this study, we attempted to improve the effectiveness of Hank's solution at preventing chorion elevation. By adjusting the concentration of the solution's contents, we developed a modified Hank's solution that can stop chorion elevation almost completely. Using this solution, we demonstrated that trypsin can induce chorion elevation. These results support our hypothesis that a trypsin-like enzyme initiates chorion elevation. This assay method can be used for the further analysis of the chorion elevation mechanism.</p>","PeriodicalId":94273,"journal":{"name":"Zebrafish","volume":"22 6","pages":"218-222"},"PeriodicalIF":1.2,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145688882","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
DaniocellDesktop: for Interactive Reanalysis of Wild-Type Zebrafish Single-Cell Genomic Data. 用于野生型斑马鱼单细胞基因组数据的交互式再分析。
IF 1.2
Zebrafish Pub Date : 2025-12-01 Epub Date: 2025-09-05 DOI: 10.1177/15458547251376181
Alicia Evans, Jeffrey A Farrell
{"title":"DaniocellDesktop: for Interactive Reanalysis of Wild-Type Zebrafish Single-Cell Genomic Data.","authors":"Alicia Evans, Jeffrey A Farrell","doi":"10.1177/15458547251376181","DOIUrl":"10.1177/15458547251376181","url":null,"abstract":"<p><p>DaniocellDesktop is a cross-platform interactive desktop application designed to facilitate reanalysis of a previously published 462,243-cell single-cell RNAseq dataset that profiled cell-type-specific gene expression across the first 5 days of wild-type zebrafish development. DaniocellDesktop enables custom redefinition of cell populations, identification of differentially expressed genes, and generation of several types of publication-ready plots to show gene expression patterns, gene co-expression patterns, and gene expression over time within these previously published data without requiring any specific programming knowledge. This software is available from https://daniocell.nichd.nih.gov/desktop/.</p>","PeriodicalId":94273,"journal":{"name":"Zebrafish","volume":" ","pages":"230-232"},"PeriodicalIF":1.2,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12976728/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145007053","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
A Simple and Cost-Effective Setup to Analyze Optokinetic Response in Adult Zebrafish. 一种分析成年斑马鱼光动力学反应的简单而经济的装置。
IF 1.2
Zebrafish Pub Date : 2025-12-01 Epub Date: 2025-09-11 DOI: 10.1177/15458547251378005
Aravindhan Gunaseelan, Keerthana Ragavi Narenthiran, M S Ananthakrishna Tantry, Kirankumar Santhakumar
{"title":"A Simple and Cost-Effective Setup to Analyze Optokinetic Response in Adult Zebrafish.","authors":"Aravindhan Gunaseelan, Keerthana Ragavi Narenthiran, M S Ananthakrishna Tantry, Kirankumar Santhakumar","doi":"10.1177/15458547251378005","DOIUrl":"10.1177/15458547251378005","url":null,"abstract":"<p><p>The optokinetic response (OKR) is a widely used measure of visual and neurological function in zebrafish (<i>Danio rerio</i>). In this study, we employed a simple and cost-effective OKR assay system for adult zebrafish utilizing a mobile phone and a stereomicroscope. The setup was validated by comparing control fish with those exposed to light-induced retinal degeneration (LIRD), demonstrating significantly impaired OKR responses. This custom-made setup offers valuable applications in visual neuroscience, disease modeling, and drug discovery.</p>","PeriodicalId":94273,"journal":{"name":"Zebrafish","volume":" ","pages":"233-236"},"PeriodicalIF":1.2,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145035052","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
Nondestructive Larval Genotyping of Danio rerio for Mitochondrial and Nuclear DNA Genetics. 鳙鱼线粒体和核DNA遗传无损分型研究。
IF 1.2
Zebrafish Pub Date : 2025-12-01 Epub Date: 2025-09-22 DOI: 10.1177/15458547251379703
Kyler S Mitra, Shannon R Holmberg, Mireya Mota, Ankit Sabharwal, Stephen C Ekker
{"title":"Nondestructive Larval Genotyping of <i>Danio rerio</i> for Mitochondrial and Nuclear DNA Genetics.","authors":"Kyler S Mitra, Shannon R Holmberg, Mireya Mota, Ankit Sabharwal, Stephen C Ekker","doi":"10.1177/15458547251379703","DOIUrl":"10.1177/15458547251379703","url":null,"abstract":"<p><p>The rapid advancement of nuclear and mitochondrial genomic editing tools has created an urgent need for efficient, nonlethal larval genotyping methods in zebrafish (<i>Danio rerio</i>) research. This study optimizes and validates a nondestructive proteinase K digestion method for mitochondrial and nuclear DNA genotyping while characterizing its impact on larval survival and gene expression. Using optimized protocol parameters, we demonstrate successful amplification of different mitochondrial and nuclear genetic loci with consistently high sensitivity. Molecular validation through PCR, restriction fragment length polymorphism analysis, and Sanger sequencing confirmed the specificity and reliability of the extracted DNA. The method successfully detected C-to-T base edits in the <i>mt-tl1</i> gene introduced using the FusX TALE Base editor system, demonstrating its applicability to gene editing studies. Both 48-well and optimized 96-well formats were used, enabling this approach to be deployed at scale. This optimized method enables researchers to correlate genotypes with phenotypes in longitudinal studies while maintaining specimen viability, particularly valuable for investigating early-onset mitochondrial diseases, and utilizes standard laboratory equipment and reagents, facilitating widespread adoption in zebrafish research while adhering to ethical principles in reducing animal mortality.</p>","PeriodicalId":94273,"journal":{"name":"Zebrafish","volume":" ","pages":"197-207"},"PeriodicalIF":1.2,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145126252","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
Effect of Variation of Spatial and Temporal Frequencies of the Moving Field Visual Stimulus on Optokinetic Response of Adult Zebrafish. 运动场视觉刺激的时空频率变化对成年斑马鱼光动力反应的影响。
IF 1.2
Zebrafish Pub Date : 2025-10-01 Epub Date: 2025-08-06 DOI: 10.1177/15458547251365847
Barnini Bhattacharya, Shibsankar Roy, Bijay Bal, Shankarashis Mukherjee, Anuradha Bhat, Kuntal Ghosh
{"title":"Effect of Variation of Spatial and Temporal Frequencies of the Moving Field Visual Stimulus on Optokinetic Response of Adult Zebrafish.","authors":"Barnini Bhattacharya, Shibsankar Roy, Bijay Bal, Shankarashis Mukherjee, Anuradha Bhat, Kuntal Ghosh","doi":"10.1177/15458547251365847","DOIUrl":"10.1177/15458547251365847","url":null,"abstract":"<p><p>The optokinetic response (OKR) is a reflexive behavior in which the eye intends to fixate the surrounding image in motion on its retina. The two important components of the OKR visual stimulus are the spatial frequency (SF) and the temporal frequency (TF). Zebrafish (<i>Danio rerio</i>) is considered an important model animal for visual physiology research. As a result, several works have been performed recently on zebrafish OKR in terms of their visual acuity or eye velocity. In the present study, a novel data pattern has been reported in terms of the relationship between ocular movement frequency (OMF) and TF of the visual stimulus in the presence of two types of SF of the optokinetic visual stimulus. The results indicated that at the basal SF (0.05 cycles per degree [cpd]), the OMF significantly increased (<i>p</i> < 0.05) with the corresponding elevation in the TF (till 240 deg.s<sup>-1</sup>). On the contrary, at a higher SF (0.29 cpd), there was a significant decrease (<i>p</i> < 0.05) in the OMF with the increase in TF.</p>","PeriodicalId":94273,"journal":{"name":"Zebrafish","volume":" ","pages":"169-174"},"PeriodicalIF":1.2,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144791165","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
A Knock-in Zebrafish Reporter Line for Live Visualization of Endogenous Olig2 Protein Dynamics. 内源性寡聚2蛋白动态可视化的敲入斑马鱼报告线。
IF 1.2
Zebrafish Pub Date : 2025-10-01 Epub Date: 2025-09-24 DOI: 10.1177/15458547251376166
Chia-Teng Chang, Toru Kawanishi, Sandy Nandagopal, Sean G Megason, Tony Y-C Tsai
{"title":"A Knock-in Zebrafish Reporter Line for Live Visualization of Endogenous Olig2 Protein Dynamics.","authors":"Chia-Teng Chang, Toru Kawanishi, Sandy Nandagopal, Sean G Megason, Tony Y-C Tsai","doi":"10.1177/15458547251376166","DOIUrl":"10.1177/15458547251376166","url":null,"abstract":"<p><p>The transcription factor oligodendrocyte transcription factor 2 (Olig2) plays a central role in specifying motor neurons and oligodendrocytes during vertebrate neural development. While transgenic reporter lines such as <i>TgBAC(olig2:EGFP)</i> have been instrumental in visualizing <i>olig2</i> expression, they fall short in directly reporting endogenous protein levels and may not fully recapitulate native gene regulation. To address these limitations, we generated a <i>TgKI(olig2-mNeonGreen)</i> zebrafish line using CRISPR/Cas9-mediated knock-in at the endogenous <i>olig2</i> locus. The resulting Olig2-mNeonGreen fusion protein localizes specifically to the nucleus, enabling direct live imaging and accurate quantification of Olig2-expressing cells. We confirmed that the knock-in preserves endogenous mRNA expression and protein function, and that homozygous fish develop normally. As proof of concept, modulation of Sonic Hedgehog signaling altered Olig2-mNeonGreen+ cell numbers as expected, confirming the reporter's responsiveness to known upstream inputs. This <i>TgKI(olig2-mNeonGreen)</i> line offers a robust tool for studying neural progenitor dynamics <i>in vivo</i>.</p>","PeriodicalId":94273,"journal":{"name":"Zebrafish","volume":" ","pages":"182-188"},"PeriodicalIF":1.2,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13090830/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145139971","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Balance Between Daily Water Renewal and Nitrate Levels in a Recirculating Zebrafish Housing System. 循环斑马鱼住宅系统中每日水更新和硝酸盐水平之间的平衡。
IF 1.2
Zebrafish Pub Date : 2025-10-01 Epub Date: 2025-08-21 DOI: 10.1177/15458547251370938
Nathan Guibert, Jean-Philippe Mocho, Laure Bally-Cuif, Sébastien Bedu
{"title":"Balance Between Daily Water Renewal and Nitrate Levels in a Recirculating Zebrafish Housing System.","authors":"Nathan Guibert, Jean-Philippe Mocho, Laure Bally-Cuif, Sébastien Bedu","doi":"10.1177/15458547251370938","DOIUrl":"10.1177/15458547251370938","url":null,"abstract":"<p><p>In zebrafish housing systems with water recirculation, a daily renewal of at least 10% of the total water volume is generally recommended to preserve optimal water quality. Such aquatic housing systems often rely on reverse osmosis (RO) water. RO production uses large amounts of running water, a high proportion of which is then discharged into waste. A key parameter for water quality control is nitrate, the final transformation product of the nitrogen cycle in water. Here, we used this parameter to test whether daily water renewal could be reduced in our zebrafish housing system without impact on water quality. For 18 months, divided into four periods, we progressively reduced the rate of water renewal from 10% to 6%. We show that nitrate levels are not impacted when renewal rates remain above 8%, making it possible to significantly save on water in zebrafish breeding systems.</p>","PeriodicalId":94273,"journal":{"name":"Zebrafish","volume":" ","pages":"189-194"},"PeriodicalIF":1.2,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144984683","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
Meeting Report: Zebrafish Education and Outreach Workshop at the 18th International Zebrafish Conference in Kyoto, Japan, on August 19, 2024. 会议报告：第18届国际斑马鱼会议斑马鱼教育和推广研讨会于2024年8月19日在日本京都举行。
IF 1.2
Zebrafish Pub Date : 2025-10-01 Epub Date: 2025-08-20 DOI: 10.1177/15458547251371863
Jennifer O Liang
{"title":"Meeting Report: Zebrafish Education and Outreach Workshop at the 18th International Zebrafish Conference in Kyoto, Japan, on August 19, 2024.","authors":"Jennifer O Liang","doi":"10.1177/15458547251371863","DOIUrl":"10.1177/15458547251371863","url":null,"abstract":"<p><p>The 2024 \"Zebrafish in Education and Outreach Workshop\" was held at the 18th International Zebrafish Conference (IZFC) in Kyoto, Japan, on August 19th. It was organized by Jennifer O. Liang of the University of Minnesota Duluth (Duluth, MN, US), with formal presentations by Jennifer Liang and Jason Meyers of Colgate University (Hamilton, NY, US), and discussion by all attendees.</p>","PeriodicalId":94273,"journal":{"name":"Zebrafish","volume":" ","pages":"195-196"},"PeriodicalIF":1.2,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144984621","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
Extraction of Ex Vivo Macrophages from the Renal Medulla of Zebrafish (Danio rerio). 斑马鱼肾髓质离体巨噬细胞的提取。
IF 1.2
Zebrafish Pub Date : 2025-10-01 Epub Date: 2025-09-24 DOI: 10.1177/15458547251366984
Leonel Witcoski Junior, André Guilherme Portela de Paula, Jordana Dinorá de Lima, Mariana Abrantes do Amaral, Barbara Nunes Padovani, Mariana Rodrigues Davanso, Niels Olsen Saraiva Camara, Karin Braun Prado, Tarcio Teodoro Braga
{"title":"Extraction of <i>Ex Vivo</i> Macrophages from the Renal Medulla of Zebrafish (<i>Danio rerio</i>).","authors":"Leonel Witcoski Junior, André Guilherme Portela de Paula, Jordana Dinorá de Lima, Mariana Abrantes do Amaral, Barbara Nunes Padovani, Mariana Rodrigues Davanso, Niels Olsen Saraiva Camara, Karin Braun Prado, Tarcio Teodoro Braga","doi":"10.1177/15458547251366984","DOIUrl":"10.1177/15458547251366984","url":null,"abstract":"<p><p>Zebrafish (Danio rerio) is recognized as a versatile model for hematopoietic studies due to its transparency, genetic similarity to humans, and ease of manipulation. This study describes a protocol for the ex vivo extraction and differentiation of macrophages from the renal medulla of zebrafish using transgenic lines and L929 cell-conditioned medium (LCCM) as an alternative to recombinant M-CSF. Adult TgMpegmCherry zebrafish were maintained under controlled conditions. Following euthanasia, renal medullary cells were isolated, treated with antibiotics, and cultured in either LCCM or recombinant human M-CSF. Macrophage differentiation was assessed using confocal microscopy (mCherry), flow cytometry, and functional phagocytosis assays. The protocol enabled efficient differentiation of progenitor cells into macrophages, with an average of 49.1% mCherry+ cells after 7 days in LCCM, outperforming recombinant M-CSF. Differentiated cells demonstrated strong phagocytic activity, confirming macrophage functionality. This method provides an accessible approach to obtaining ex vivo zebrafish-derived macrophages, enabling immunological and hematopoietic studies and allowing for functional comparisons with traditional murine macrophage protocols.</p>","PeriodicalId":94273,"journal":{"name":"Zebrafish","volume":" ","pages":"175-181"},"PeriodicalIF":1.2,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145139981","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
A Standardized Structural Environmental Enrichment for Zebrafish Kept Research Facilities. 斑马鱼养殖研究设施的标准化结构环境富集。
IF 1.2
Zebrafish Pub Date : 2025-08-01 Epub Date: 2025-06-30 DOI: 10.1089/zeb.2025.0024
Wendela Vester, Karin Pernold, Alexander Wikström, Lars Bräutigam
{"title":"A Standardized Structural Environmental Enrichment for Zebrafish Kept Research Facilities.","authors":"Wendela Vester, Karin Pernold, Alexander Wikström, Lars Bräutigam","doi":"10.1089/zeb.2025.0024","DOIUrl":"10.1089/zeb.2025.0024","url":null,"abstract":"<p><p>Environmental enrichment for research animals does not only promote animal welfare but is also essential for obtaining scientifically relevant and reproducible results. Currently, all available \"enrichments\" for zebrafish aquaria are either not standardized, pose a biosafety risk, or may lead to increased aggression due to their monopolization by the dominant individual. We have developed the environmental enrichment divider which provides standardized enrichment for zebrafish aquaria mimicking hanging vegetation found in the natural habitat of zebrafish. The divider cannot be monopolized and thereby does not lead to increased aggression. Furthermore, it can easily be implemented in the workflow of any zebrafish research facility and is made of inert, autoclavable plastic, which does not pose a biosafety risk.</p>","PeriodicalId":94273,"journal":{"name":"Zebrafish","volume":" ","pages":"152-157"},"PeriodicalIF":1.2,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144532223","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
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