Zebrafish Pub 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。

Zebrafish Pub 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":"https://doi.org/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":""},"PeriodicalIF":0.0,"publicationDate":"2025-06-11","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

Anxiety and Fryes: A Novel Model to Study Anxiety in Zebrafish Larvae. 焦虑与焦虑：斑马鱼幼虫焦虑研究的新模型。

Zebrafish Pub Date : 2025-06-01 Epub Date: 2025-05-15 DOI: 10.1089/zeb.2025.0002

Ana Cláudia Costa de Carvalho, Sílvio Morato, Amauri Gouveia, André Walsh-Monteiro

{"title":"Anxiety and Fryes: A Novel Model to Study Anxiety in Zebrafish Larvae.","authors":"Ana Cláudia Costa de Carvalho, Sílvio Morato, Amauri Gouveia, André Walsh-Monteiro","doi":"10.1089/zeb.2025.0002","DOIUrl":"10.1089/zeb.2025.0002","url":null,"abstract":"Over the last few decades, zebrafish have proven to be a valuable tool for biomedical research, with advantages mainly in the use of embryos and adults. The larval stage, on the contrary, is somewhat less used, generally due to a lack of protocols compared with the other stages. In this study, we propose a protocol to study anxiety-like behavior in larvae using the plus maze with ramp (PMR). In the PMR, anxious behavior is measured by the time spent on the flat arms relative to the ramped arms of the apparatus. In the first phase of the study, animals at 5, 14, and 21 days postfertilization (dpf) were exposed to the PMR at different water column heights and session times. It was observed that animals at 14 and 21 dpf were more sensitive to the PMR and exhibited obvious anxiety-like behavior compared with larvae at 5 dpf. In the second phase of the study, to assess the robustness of the type-anxious response exhibited by the larvae, animals at 5, 10, and 14 dpf treated with anxiolytic drugs were exposed to PMR. Alcohol exposure showed a bimodal effect for animals at 10 and 14 dpf, with an anxiolytic effect at intermediate doses, with more exploration of the apparatus and time in the ramp arms. Higher doses caused a reduction in locomotor activity characteristic of drunkenness. Exposure to clonazepam produced anxiolytic effects and a reduction in locomotor activity at the highest dose in 10 and 14 dpf animals. On the contrary, 5 dpf animals showed unexpected effects, which we believe to be related to their stage of development. The results indicate that the PMR is an effective tool for assessing anxiety-like behavior in zebrafish larvae and is sensitive to anxiolytic drugs. Furthermore, anxiety-like behavior appears to vary during the early stages of larval development.","PeriodicalId":94273,"journal":{"name":"Zebrafish","volume":" ","pages":"86-98"},"PeriodicalIF":0.0,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144083122","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

How to Dissect Zebrafish: A Standardized Sampling Protocol for Histological and Molecular Studies in Adult Zebrafish. 如何解剖斑马鱼：成年斑马鱼组织学和分子研究的标准化采样协议。

Zebrafish Pub Date : 2025-06-01 Epub Date: 2025-04-26 DOI: 10.1089/zeb.2024.0184

Martín Pérez Díaz, Ana Manuela de Azevedo Gomes, María Isabel Quiroga Berdeal, Roberto Bermúdez Pose

{"title":"How to Dissect Zebrafish: A Standardized Sampling Protocol for Histological and Molecular Studies in Adult Zebrafish.","authors":"Martín Pérez Díaz, Ana Manuela de Azevedo Gomes, María Isabel Quiroga Berdeal, Roberto Bermúdez Pose","doi":"10.1089/zeb.2024.0184","DOIUrl":"10.1089/zeb.2024.0184","url":null,"abstract":"The zebrafish (Danio rerio) has become an essential model in fields such as developmental biology, toxicology, genetics, and regenerative medicine due to its low cost, small size, transparent embryos, and genetic similarity to humans. Nowadays, this model is increasingly recognized as a valuable tool in other fields, including veterinary medicine and animal production research, particularly aquaculture, due to its unique characteristics that make it suitable for studying economically significant diseases affecting production species. However, unlike established models such as mice, zebrafish lack standardized protocols for housing, feeding, anesthesia, and sample collection, which affects study reproducibility. In addition, it is a common practice to use whole zebrafish larvae or juveniles for metagenomic studies rather than analyzing individual organs, despite the fact that gene expression can vary between organs. This approach complicates the attribution of findings to specific biological processes. To address this, various authors proposed protocols for sample collection in larvae, juveniles, and adult zebrafish; however, comprehensive studies encompassing nearly all fish organs are scarce. Understanding zebrafish anatomy and the technical requirements of the study is essential for accurate sample collection. Some challenges present during zebrafish dissection include the small size of the fish, the fragility of their organs, and the faster onset of autolysis and heterolysis after the death of the animal. Using magnifying lenses, microdissection tools, and conducting dissections on cold surfaces can help mitigate these issues. This article aims to improve sample collection for histopathological and genetic studies in adult zebrafish by establishing a comprehensive, organized, and systematic dissection protocol that accounts for the anatomical specifics of this experimental model.","PeriodicalId":94273,"journal":{"name":"Zebrafish","volume":" ","pages":"76-85"},"PeriodicalIF":0.0,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144049414","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

Innovative Approaches in Developmental Biology: Integrating Zebrafish as a Model Organism in Education. 发展生物学的创新方法：将斑马鱼作为教育中的模式生物。

Zebrafish Pub Date : 2025-06-01 Epub Date: 2025-05-22 DOI: 10.1089/zeb.2024.0182

Magda Dubińska-Magiera, Ewa Stefanik, Marta Migocka-Patrzałek

引用次数: 0

Comprehensive 3D Imaging of Whole Zebrafish Using a Water-Based Clearing Reagent for Hard Tissues. 利用水基硬组织清除试剂对整个斑马鱼进行全面的三维成像。

Zebrafish Pub Date : 2025-06-01 DOI: 10.1089/zeb.2025.0018

Kiyohito Taimatsu, John Prevedel, Daniel Castranova, Kanako Inoue, Amit Puthan, Brant M Weinstein

{"title":"Comprehensive 3D Imaging of Whole Zebrafish Using a Water-Based Clearing Reagent for Hard Tissues.","authors":"Kiyohito Taimatsu, John Prevedel, Daniel Castranova, Kanako Inoue, Amit Puthan, Brant M Weinstein","doi":"10.1089/zeb.2025.0018","DOIUrl":"10.1089/zeb.2025.0018","url":null,"abstract":"Zebrafish (Danio rerio) is a valuable model organism for studying developmental processes due to its external development and the optical clarity of its embryos and larvae. However, as development proceeds, zebrafish form increasingly opaque tissues that impede visualization of deep tissues and structures. Although tissue clearing methods have been used to facilitate imaging at these later stages, most of these methods have limited ability to clear dense tissues such as bone and cartilage, cause significant morphological distortion, and/or result in loss of fluorescent signal when used for imaging of fluorescent transgenes, dye-stained animals, or specimens generated using immunofluorescence or fluorescence in situ hybridization methods. Here, we report a novel imaging technique using a recently developed clearing reagent called LUCID that makes it possible to capture the complete cellular-resolution 3D structures of larval and juvenile zebrafish. We show that LUCID clears dense tissues such as pharyngeal cartilage in juvenile animals and even tooth bone in adults without causing either significant morphological distortion or significant loss of signal from transgene-driven fluorescent proteins, fluorescent nuclear DNA or actin staining dyes, or whole-mount in situ hybridization chain reaction fluorescence. Using this new approach, it is possible to perform complete high-resolution 3D imaging of whole fluorescently stained animals, even deep internal regions, providing a novel tool for elucidating the complex internal structures of developing zebrafish.","PeriodicalId":94273,"journal":{"name":"Zebrafish","volume":"22 3","pages":"65-75"},"PeriodicalIF":0.0,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144287705","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

Evaluation of Tricaine Methanesulfonate Concentrations for Flow Anesthesia in Zebrafish (Danio rerio). 斑马鱼流动麻醉中三卡因甲磺酸浓度的评价。

Zebrafish Pub Date : 2025-06-01 Epub Date: 2025-04-23 DOI: 10.1089/zeb.2025.0005

Alexander G Kramer, Logan R Bern, Lindsey T Ferguson, Michael B Palillo, Raphael A Malbrue, Mary E White, Genevieve C Kendall, Carmen B Arsuaga-Zorrilla, Logan A Fehrenbach

{"title":"Evaluation of Tricaine Methanesulfonate Concentrations for Flow Anesthesia in Zebrafish (Danio rerio).","authors":"Alexander G Kramer, Logan R Bern, Lindsey T Ferguson, Michael B Palillo, Raphael A Malbrue, Mary E White, Genevieve C Kendall, Carmen B Arsuaga-Zorrilla, Logan A Fehrenbach","doi":"10.1089/zeb.2025.0005","DOIUrl":"10.1089/zeb.2025.0005","url":null,"abstract":"Zebrafish (Danio rerio) are often anesthetized by immersion in buffered tricaine methanesulfonate (MS-222). Although commonly utilized, immersion anesthesia presents a shortcoming of lethal asphyxiation with increased duration of exposures. A newer technique that circumvents this issue, known as flow anesthesia, has been adapted from larger aquatic species to zebrafish. Flow anesthesia improves safety by delivering oxygen-rich water along with the anesthetic across gill epithelium and allowing fish to be manipulated outside of water. Information on the construction of flow anesthesia apparatuses and parameters are sparse. The goal of this study was to create a flow anesthesia apparatus with materials commonly found within a research facility and to evaluate variable concentrations of MS-222 for anesthesia in zebrafish. Depth of anesthesia was monitored by quantifying respiratory rate and recording responses to physical stimulation. All concentrations of MS-222 evaluated (30-100 ppm) were successful at maintaining surgical anesthesia for up to 30 min. The anesthetic events were demonstrated to be safe, with an observed 97% survival rate. This work demonstrates refinements in zebrafish anesthesia and encourages future studies to evaluate MS-222 flow anesthesia for longer duration and evaluation of other commercially available anesthetics for efficacy in a flow anesthesia setup.","PeriodicalId":94273,"journal":{"name":"Zebrafish","volume":" ","pages":"99-106"},"PeriodicalIF":0.0,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144001201","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

Understanding Rigor Mortis Impacts on Zebrafish Gamete Viability. 了解僵死对斑马鱼配子活力的影响。

Zebrafish Pub Date : 2025-04-01 Epub Date: 2025-02-27 DOI: 10.1089/zeb.2024.0177

Renata Villar Dantas, Lis Santos Marques, Thaiza Rodrigues Freitas, Nathalia Dos Santos Teixeira, Rômulo Batista Rodrigues, Jhony Lisbôa Benato, Raquel Santos Dos Santos, Diógenes Henrique Siqueira Silva, Danilo Pedro Streit

{"title":"Understanding Rigor Mortis Impacts on Zebrafish Gamete Viability.","authors":"Renata Villar Dantas, Lis Santos Marques, Thaiza Rodrigues Freitas, Nathalia Dos Santos Teixeira, Rômulo Batista Rodrigues, Jhony Lisbôa Benato, Raquel Santos Dos Santos, Diógenes Henrique Siqueira Silva, Danilo Pedro Streit","doi":"10.1089/zeb.2024.0177","DOIUrl":"10.1089/zeb.2024.0177","url":null,"abstract":"This study aimed to evaluate the viability of gametes in zebrafish (Danio rerio), at different rigor mortis stages. Viability assessments were conducted on oocytes at various developmental stages using LIVE/DEAD and the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction assay. For sperm evaluation, both kinetic (computer-assisted sperm analysis) and morphological assessments (Rose Bengal staining) were performed. Results demonstrated that rigor mortis progression significantly impacted oocyte viability during post-rigor stages, with the following viability rates: pre-rigor (70.43 ± 12.31%), fresh/control (46.43 ± 12.54%), post-rigor (27.62 ± 22.29%), and rigor mortis (comparable to fresh/control). Conversely, sperm kinetics exhibited nuanced responses to the rigor mortis stages, with specific parameters showing sensitivity, whereas the others remained relatively stable. Sperm motility was higher in the fresh/control (63.23 ± 19.03%) and pre-rigor (58.96 ± 14.38%) compared to the post-rigor group (3.34 ± 4.65%). This study highlights the significance of the pre-rigor for successful gamete collection and preservation. These findings provide valuable insights for conservation efforts and optimization of genetic resource management for endangered fish species. This study aimed to develop effective assistive reproductive techniques by elucidating the interplay between rigor mortis and gamete quality, contributing to the broader goals of species conservation and maintenance of genetic diversity in fish populations.","PeriodicalId":94273,"journal":{"name":"Zebrafish","volume":" ","pages":"31-41"},"PeriodicalIF":0.0,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143525759","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

VMS-Fish: A Comprehensive Application for Zebrafish Husbandry Management. VMS-Fish：斑马鱼养殖管理的综合应用。

Zebrafish Pub Date : 2025-04-01 Epub Date: 2025-04-08 DOI: 10.1089/zeb.2024.0183

Vishnu Muraleedharan Saraswathy, Mayssa H Mokalled

引用次数: 0

An Improved, High-Yield Method for Isolating Nuclei from Individual Zebrafish Embryos for Single-Nucleus RNA Sequencing. 一种改进的、高产的从单个斑马鱼胚胎中分离细胞核进行单核RNA测序的方法。

Zebrafish Pub Date : 2025-04-01 Epub Date: 2025-03-05 DOI: 10.1089/zeb.2024.0175

Clifford Rostomily, Heidi Lee, Amy Tresenrider, Riza Daza, Andrew Mullen, Jay Shendure, David Kimelman, Cole Trapnell

引用次数: 0

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