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

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":"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 TgBAC(olig2:EGFP) have been instrumental in visualizing olig2 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 TgKI(olig2-mNeonGreen) zebrafish line using CRISPR/Cas9-mediated knock-in at the endogenous olig2 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 TgKI(olig2-mNeonGreen) line offers a robust tool for studying neural progenitor dynamics in vivo.","PeriodicalId":94273,"journal":{"name":"Zebrafish","volume":" ","pages":"182-188"},"PeriodicalIF":1.2,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145139971","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

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

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

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 Ex Vivo Macrophages from the Renal Medulla of Zebrafish (Danio rerio).","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":"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.","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

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 : 2025-09-22 DOI: 10.1177/15458547251379703

Kyler S Mitra, Shannon R Holmberg, Mireya Mota, Ankit Sabharwal, Stephen C Ekker

{"title":"Nondestructive Larval Genotyping of Danio rerio 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":"The rapid advancement of nuclear and mitochondrial genomic editing tools has created an urgent need for efficient, nonlethal larval genotyping methods in zebrafish (Danio rerio) 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 mt-tl1 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.","PeriodicalId":94273,"journal":{"name":"Zebrafish","volume":" ","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-09-22","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

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 : 2025-09-11 DOI: 10.1177/15458547251378005

Aravindhan Gunaseelan, Keerthana Ragavi Narenthiran, M S Ananthakrishna Tantry, Kirankumar Santhakumar

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 : 2025-09-05 DOI: 10.1177/15458547251376181

Alicia Evans, Jeffrey A Farrell

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

引用次数: 0

A Novel Method to Make Zebrafish Mate by Adding an Membrane Progestin Receptor Agonist to the Water Without Injection. 在水中不注射添加膜孕激素受体激动剂使斑马鱼交配的新方法。

IF 1.2

Zebrafish Pub Date : 2025-08-01 Epub Date: 2025-06-11 DOI: 10.1089/zeb.2025.0020

Saokat Ahamed, Md Abul Bashar, Mohammad Maksudul Hassan, Toshinobu Tokumoto

{"title":"A Novel Method to Make Zebrafish Mate by Adding an Membrane Progestin Receptor Agonist to the Water Without Injection.","authors":"Saokat Ahamed, Md Abul Bashar, Mohammad Maksudul Hassan, Toshinobu Tokumoto","doi":"10.1089/zeb.2025.0020","DOIUrl":"10.1089/zeb.2025.0020","url":null,"abstract":"This study investigates a novel method to induce mating in zebrafish (Danio rerio) by direct water addition of membrane progestin receptor (mPR) agonists, Org OD 02-0 (Org), or a maturation-inducing steroid, 17α,20β-dihydroxy-4-pregnen-3-one (DHP). Traditional spawning induction methods using hormone injections can be invasive and stressful for fish, especially small fish such as zebrafish. A previous study showed that progestins applied externally to the water can induce oocyte maturation and ovulation in female zebrafish. Then, a recent study showed that the addition of steroids and Org to the water also induced male sexual behavior in zebrafish. Therefore, it was hypothesized that the addition of these compounds to the water could induce mating in zebrafish. To address this, we investigated the efficacy of adding these compounds to the water in the vessel with a pair of zebrafish. Female and male zebrafish pairs were exposed to varying concentrations of DHP or Org in the water, and the resulting spawning rates and rate of sexual behavior induction were recorded. Our results showed that both simple addition of DHP and Org solution successfully induced mating within 4 h. A single chemical induced both spawning in females and sexual behavior in males. In this study, we have established a simple, novel method to induce mating in zebrafish by adding mPR agonists to the water.","PeriodicalId":94273,"journal":{"name":"Zebrafish","volume":" ","pages":"136-140"},"PeriodicalIF":1.2,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144268323","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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