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

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

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 : 2025-05-15 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":"https://doi.org/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":""},"PeriodicalIF":0.0,"publicationDate":"2025-05-15","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 : 2025-04-26 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":"https://doi.org/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":""},"PeriodicalIF":0.0,"publicationDate":"2025-04-26","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

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 : 2025-04-23 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":"https://doi.org/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":""},"PeriodicalIF":0.0,"publicationDate":"2025-04-23","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

Small Fishes, Big Issues: Species Delimitation in Hemigrammus Marginatus, Gill, 1958 (Acestrorhamphidae: Pristellinae) from Brazilian Coastal Basins Based on Integrative Genetics. 小鱼类，大问题：基于整合遗传学的巴西沿海流域 Hemigrammus Marginatus, Gill, 1958 (Acestrorhamphidae: Pristellinae) 鱼种划分。

Zebrafish Pub Date : 2025-04-01 Epub Date: 2025-02-14 DOI: 10.1089/zeb.2024.0174

Mauricio B Fernandes, Jamille de Araújo Bitencourt, André Teixeira da Silva, Marcelo Ricardo Vicari, Matheus Azambuja, Paulo Roberto Antunes de Mello Affonso

{"title":"Small Fishes, Big Issues: Species Delimitation in Hemigrammus Marginatus, Gill, 1958 (Acestrorhamphidae: Pristellinae) from Brazilian Coastal Basins Based on Integrative Genetics.","authors":"Mauricio B Fernandes, Jamille de Araújo Bitencourt, André Teixeira da Silva, Marcelo Ricardo Vicari, Matheus Azambuja, Paulo Roberto Antunes de Mello Affonso","doi":"10.1089/zeb.2024.0174","DOIUrl":"10.1089/zeb.2024.0174","url":null,"abstract":"The small characins represent a systematic puzzle in the Neotropical ichthyofauna as a result of independent miniaturization processes, adaptive convergence and lack of diagnostic characters for several genera. In order to diminish the taxonomic uncertainties and the evolutionary pathways in Hemigrammus, we carried out an integrative genetic analysis in the putatively widespread Hemigrammus marginatus Ellis, 1958 by combining cytogenetic and molecular data based on the mitochondrial Cytochrome C Oxidase subunit I (COI). Specimens of H. marginatus from the type locality in Itapicuru River basin and other two populations from coastal rivers in northeastern Brazil were analyzed and compared with the available data from other regions in South America. Conspicuous macro and microkaryotypic differences were detected between the samples from northeastern and southern Brazil (Upper Paraná River basin). Likewise, the DNA barcoding and species delimitation analyses recovered distinct Molecular Operational Taxonomical Units within H. marginatus. Therefore, the population from the type locality should be referred to as H. marginatus stricto sensu, representing a restricted characin taxon from coastal drainages (including the São Francisco River basin) along northeastern Brazil, while other populations of this small characin fish need to be taxonomically revised and managed as unique lineages.","PeriodicalId":94273,"journal":{"name":"Zebrafish","volume":" ","pages":"46-58"},"PeriodicalIF":0.0,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143417133","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 Novel Food Delivery Method for Learning Studies Detects Significant Differences in Food Preference in Zebrafish. 一种用于学习研究的新型食物传递方法检测斑马鱼食物偏好的显着差异。

Zebrafish Pub Date : 2025-02-01 Epub Date: 2025-02-06 DOI: 10.1089/zeb.2024.0176

Amira Abozaid, Melissa Scaramuzzo, Hemali Patel, Robert Gerlai

{"title":"A Novel Food Delivery Method for Learning Studies Detects Significant Differences in Food Preference in Zebrafish.","authors":"Amira Abozaid, Melissa Scaramuzzo, Hemali Patel, Robert Gerlai","doi":"10.1089/zeb.2024.0176","DOIUrl":"10.1089/zeb.2024.0176","url":null,"abstract":"Despite decades of research with laboratory rodents, the mechanisms underlying learning and memory, and their impairment, are still not fully understood. The zebrafish is a newcomer in this research area, but it has shown great promise. Food is often employed as a reinforcer in learning tasks with rodents. However, for zebrafish, food has been a problematic reinforcer. Controlling timing and localization of its delivery is difficult. What food types zebrafish prefer is also rarely studied? Here, we describe a novel food delivery hardware and procedure. The apparatus is simple, cheap to manufacture, and easy to employ. Using this new method, we compare how zebrafish respond to three food types, artemia nauplii, crushed tropical fish flakes, and small zebrafish pellets. In binary choice tasks, we show that zebrafish spend significantly more time near the artemia delivery cylinder, swim closer to, and visit this cylinder more frequently compared to food cylinders delivering flakes or pellets, while responses to these latter two cylinders do not differ from each other. We conclude that the newly developed method allows the quantification of food preference in zebrafish, and that it will lead to the identification of highly rewarding food types for learning studies in this species.","PeriodicalId":94273,"journal":{"name":"Zebrafish","volume":" ","pages":"20-29"},"PeriodicalIF":0.0,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143257741","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

Acknowledgment of Reviewers 2024. 审稿人致谢

Zebrafish Pub Date : 2025-02-01 DOI: 10.1089/zeb.2024.58862.revack

引用次数: 0

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