Zebrafish最新文献

{"title":"The Need for a Framework Addressing the Temporal Aspects of Fish Sperm Motility Leading to Community-Level Standardization.","authors":"Harvey Blackburn,&nbsp;Leticia Torres,&nbsp;Yue Liu,&nbsp;Terrence R Tiersch","doi":"10.1089/zeb.2022.0006","DOIUrl":"https://doi.org/10.1089/zeb.2022.0006","url":null,"abstract":"<p><p>Motility is a widely available parameter that can be used to assess sperm quality of aquatic species. Sperm from fishes with external fertilization usually undergo a dynamic and short-lived period of motility after activation. The common practice of assigning a single value at an arbitrary peak of motility presents challenges for reproducibility, community-level standardization, and comparisons across studies. This study aimed to explore statistical approaches to standardize motility reporting, and to develop an initial framework for community-level standards. Sperm samples from 14 zebrafish (<i>Danio rerio</i>) with a total of 21,705 cells were analyzed by use of computer-assisted sperm analysis with data collection starting at 10 s after activation at 5-s intervals for 50 s. Four common motility variables were selected for analyses: curvilinear velocity, straight-line velocity, beat cross frequency, and amplitude of lateral head displacement. Cluster analysis was used to evaluate sperm subpopulations within and among males over time, least-square means was used to explore temporal aspects, and the first derivative of the regression equations was used to calculate the rate of change for the motility parameters. Cluster analysis proved informative, but overlapping ephemeral clusters were not valuable for providing standardization options. Analysis of temporal aspects and rate of change indicated opportunities for standardization by reporting the overall motility-time functions or reporting during stable time windows instead of peak motility or at random times. These approaches could minimize the inconsistencies caused by male-to-male variation and dynamic changes of subpopulations while providing comparable information. An overall temporal framework was identified for motility reporting along the collection-processing-cryopreservation-thawing sequence to provide a basis to support efforts of community-level standardization.</p>","PeriodicalId":23872,"journal":{"name":"Zebrafish","volume":null,"pages":null},"PeriodicalIF":2.0,"publicationDate":"2022-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9419943/pdf/zeb.2022.0006.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10290576","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Anesthesia Overdose Versus Rapid Cooling for Euthanasia of Adult Zebrafish.","authors":"Jorge M Ferreira,&nbsp;Luís Félix,&nbsp;Sara Jorge,&nbsp;Sandra M Monteiro,&nbsp;I Anna S Olsson,&nbsp;Ana M Valentim","doi":"10.1089/zeb.2022.0001","DOIUrl":"https://doi.org/10.1089/zeb.2022.0001","url":null,"abstract":"<p><p>The rapid increase in zebrafish use needs to be accompanied by research into the refinement of procedures. The European (EU) Directive lists three possible euthanasia methods for fish: anesthetic overdose, electrical stunning, and concussion. However, for small fish such as zebrafish, concussion and electrical stunning are difficult to perform, leaving anesthetic overdose as the most used method. Our aim was to test the efficacy and side effects of anesthesia overdose using different anesthetics and the rapid cooling method to euthanize adult zebrafish. Adult mixed-sex AB zebrafish were randomly assigned to: 250 mg/L MS222; 20 mg/L propofol +100 mg/L lidocaine; 6 mg/L etomidate; 50 mg/L clove oil; and rapid cooling (water at 2°C-4°C). Two minutes after opercular movement ceased, animals were transferred into clean water for 20 min and recovery assessed, or decapitated and used for biochemical analysis of the gills, muscle, liver, and brain; for the histological analysis of the gills and muscle; or for the assessment of cortisol levels. No animal recovered; rapid cooling was the quickest and etomidate overdose was the slowest method to cease the opercular movements. There were no major differences between euthanasia methods regarding the biochemical or histological data. Cortisol levels were higher in the rapid cooling group, but only when compared with the propofol/lidocaine group. The use of a physical method of euthanasia, such as rapid cooling, is essential when chemicals, such as anesthetics, may interfere with postmortem analyses. Although anesthetic overdose can be used without major effects on the analyses conducted in this work, rapid cooling can be another option with the advantage of being simple to administer, easily available, affordable, and very quick; this decreases the potential duration of suffering, being more humane. Therefore, a change in EU legislation should be considered to include additional humane options for euthanasia, such as rapid cooling, for zebrafish and other small tropical fish.</p>","PeriodicalId":23872,"journal":{"name":"Zebrafish","volume":null,"pages":null},"PeriodicalIF":2.0,"publicationDate":"2022-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40404252","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Rapid and Noninvasive Method That Extracts Polymerase Chain Reaction-Ready Genomic DNA from Adult Zebrafish.","authors":"Ji Yun Jang,&nbsp;Ting Liang,&nbsp;Myeong-Kyu Kim,&nbsp;Kyung Wook Kang,&nbsp;Bora Lee,&nbsp;Seok-Yong Choi","doi":"10.1089/zeb.2022.0015","DOIUrl":"https://doi.org/10.1089/zeb.2022.0015","url":null,"abstract":"<p><p>Genotyping usually entails analysis of the products of polymerase chain reaction (PCR) carried out with genomic DNA (gDNA) as template, and is employed for validation of mutant or transgenic organisms. For genotyping of adult zebrafish, gDNA is often extracted from clipped caudal fin or skin mucus through either alkaline lysis using NaOH or proteinase K (PK) treatment. Further purification of the gDNA using ethanol precipitation was optional. To develop a rapid and noninvasive method that extracts PCR-ready gDNA from adult zebrafish, we combined skin swabbing with PK treatment and demonstrated its efficiency. This method could be applied to a wide range of fish.</p>","PeriodicalId":23872,"journal":{"name":"Zebrafish","volume":null,"pages":null},"PeriodicalIF":2.0,"publicationDate":"2022-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9419989/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40622492","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Simple and High-Throughput Rheotaxis Behavioral Assay for Zebrafish Larva.","authors":"M. Tantry, V. S. Harini, Kirankumar Santhakumar","doi":"10.1089/zeb.2021.0087","DOIUrl":"https://doi.org/10.1089/zeb.2021.0087","url":null,"abstract":"Zebrafish (Danio rerio) is used as a model for studying sensorineural hearing loss. The damage to the hair cells can be assessed by scoring rheotaxis behavior in zebrafish. In this study, we newly designed a rheotaxis behavioral assay protocol capable of quantifying rheotaxis behavior in zebrafish larvae. We chemically induced ototoxicity in the larvae using copper sulfate, a well-known ototoxin, and determined rheotaxis at different flow velocities. The simple, cost-effective, and high-throughput rheotaxis assay system can provide great insights into drug development and other behavioral studies.","PeriodicalId":23872,"journal":{"name":"Zebrafish","volume":null,"pages":null},"PeriodicalIF":2.0,"publicationDate":"2022-06-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43595079","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification of Transcription Factor Binding Sites by Cleavage Under Target and Release Using Nuclease in Zebrafish","authors":"Radwa A. Barakat, C. Campbell, Raquel Espín-Palazón","doi":"10.1089/zeb.2021.0082","DOIUrl":"https://doi.org/10.1089/zeb.2021.0082","url":null,"abstract":"Cleavage Under Targets and Release Using Nuclease (CUT&RUN) has emerged as a chromatin profiling strategy that excels traditional methods. Although CUT&RUN has been widely utilized in mammalian cells, its use in the zebrafish is at its early stages. In this study, we have developed a protocol to successfully perform CUT&RUN to map transcription factor (TF) binding sites in embryonic, adult tissues, and FACS-sorted zebrafish cells. We also provide a detailed workflow for the identification of predicted TF binding sites that can be utilized in any animal species. Altogether, our strategy will expand this invaluable tool to the zebrafish community, improving the epigenetic resolution that can be achieved in this model organism.","PeriodicalId":23872,"journal":{"name":"Zebrafish","volume":null,"pages":null},"PeriodicalIF":2.0,"publicationDate":"2022-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47974717","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Feeding Rate and Protein Quality Differentially Affect Growth and Feeding Efficiency Response Variables of Zebrafish (<i>Danio rerio</i>).","authors":"Shaley Valentine, Karolina Kwasek","doi":"10.1089/zeb.2022.0002","DOIUrl":"10.1089/zeb.2022.0002","url":null,"abstract":"<p><p>Manipulating feeding rate and protein quality may improve growth and feeding efficiency of cultured species. However, whether feeding rate, protein quality, or their interaction has a greater effect on growth and feeding efficiency response variables is unknown. To determine whether feeding rate and protein quality individually or interactively affect growth and feeding efficiency, juvenile Zebrafish (<i>Danio rerio</i>) were either offered nutritionally similar diet consisting of either menhaden fishmeal protein or a 100% replacement of fishmeal with soybean meal-based protein restrictively or to satiation. Total length, weight, feed intake, and feed conversion ratio (FCR) were measured throughout the duration of the study. Protein quality and feeding rate individually and interactively affected feed intake and FCR: Zebrafish offered feed to satiation had higher growth and FCR than those fed restrictively, and Zebrafish fed soybean meal-based diet showed lower growth and higher FCR and feed intake compared to those fed fishmeal-based diet, although magnitude of response depended on feeding rate. These findings likely indicate lower digestibility of soybean meal or the presence of antinutritional factors in soybean meal that led to impaired nutrient absorption of fish offered soybean meal-based diet. Differences in measured response variables between protein qualities and feeding rates highlight the importance of determining interactive effects in nutritional studies.</p>","PeriodicalId":23872,"journal":{"name":"Zebrafish","volume":null,"pages":null},"PeriodicalIF":2.0,"publicationDate":"2022-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44239097","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Dopamine Level Affects Social Interaction and Color Preference Possibly Through Intestinal Microbiota in Zebrafish.","authors":"Ju Wang, Feng Zheng, Lifen Yin, Shengnan Shi, Bing Hu, Hao Qu, Lei Zheng","doi":"10.1089/zeb.2021.0077","DOIUrl":"https://doi.org/10.1089/zeb.2021.0077","url":null,"abstract":"Accumulating researches suggest that the microbiota reside in the gastrointestinal system can influence neurodevelopment of brain and programming of behaviors. However, the mechanism underlining the relationship between shoals' behaviors and intestinal microbiota remain controversial and the roles of responsible neurotransmitters are still unclear. Here we show that shoaling behavior affected the color preference of shoals, indicating that shoals tended to choose a favorable color environment that benefited social contact. Meanwhile, administration of the selective D1-R antagonist, SCH23390, could disrupt the social interaction that led to the deficits of color preference in shoals. More importantly, the altered microbiota caused by an antibiotic oxytetracycline (OTC) exposure decreased the sociability and weakened shoals' preference for all color combinations. When given a supplementation of Lactobacillus rhamnosus GG after OTC exposure, fish maintained the same capability of social cohesion and color preference as normal fish. Our results support a role for dopamine in shaping the color preference in shoals. Our findings show that dopamine level of brain could mediate both social recognition and color preference, and offer a possibility that the production of dopamine is coordinated through gut microbiota.","PeriodicalId":23872,"journal":{"name":"Zebrafish","volume":null,"pages":null},"PeriodicalIF":2.0,"publicationDate":"2022-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47343979","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A New Transgenic Line for Rapid and Complete Neutrophil Ablation.","authors":"Christopher J Hall,&nbsp;Jonathan W Astin,&nbsp;Jeff S Mumm,&nbsp;David F Ackerley","doi":"10.1089/zeb.2022.0020","DOIUrl":"https://doi.org/10.1089/zeb.2022.0020","url":null,"abstract":"<p><p>Zebrafish lines expressing nitroreductase (NTR) in specific cell compartments, which sensitizes those cells to metronidazole (MTZ)-mediated ablation, have proven extremely useful for studying tissue regeneration and investigating cell function. In contrast to many cells, neutrophils are comparatively resistant to the NTR/MTZ targeted ablation strategy. Recently, a rationally engineered variant of NTR (NTR 2.0) has been described that exhibits greatly improved MTZ-mediated ablation efficacy in zebrafish. We show that a transgenic line with neutrophil-restricted expression of NTR 2.0 demonstrates complete neutrophil ablation, with an MTZ dose 100-fold less than current treatment regimens, and with treatment durations as short as 5 h.</p>","PeriodicalId":23872,"journal":{"name":"Zebrafish","volume":null,"pages":null},"PeriodicalIF":2.0,"publicationDate":"2022-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9246267/pdf/zeb.2022.0020.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9556890","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"TP53 Mutant Versus Wild-Type Zebrafish Larvae Under Starvation Stress: Larvae Can Live Up to 17 Days Post-Fertilization Without Food.","authors":"K. Voskarides, Constantina Koutsofti, Maria Pozova","doi":"10.1089/zeb.2022.0003","DOIUrl":"https://doi.org/10.1089/zeb.2022.0003","url":null,"abstract":"In this study, an experimental protocol has been developed for comparing survival rates of mutant and wild-type zebrafish larvae under extreme starvation. Zebrafish larvae were placed in 96-well plates at fourth day postfertilization (dpf) and larvae were not fed at all from hatching to cease. Zdf1 zebrafish line was used, a strain carrying the (cancer) pathogenic TP53-M214K amino acid substitution. TP53-M214 corresponds to the human TP53-M246 and both residues are located on the DNA-binding domain of the p53 protein. Survival statistical analysis did not show any significant difference in the overall survival rates between homozygous mutant and wild-type larvae. When considering 15 dpf as the endpoint of the experiment (66% of larvae died), a borderline statistical significance was observed for the dominant model of inheritance (p = 0.015; relative hazard = 0.8320). Despite the fact yolk sac of larvae is depleted at 7-8 dpf, 34% of larvae survive until 15 dpf and 1.5% until 17 dpf. Concluding, three main results derive from this study: (1) pathogenic homozygous mutations in TP53 probably do not alter survival rates of zebrafish larvae under starvation; (2) zebrafish larvae can live up to 17 dpf without food, surviving only with their initial nutritional supplies; and (3) an easy and affordable protocol has been developed for estimating survival rates of zebrafish larvae under stressful conditions.","PeriodicalId":23872,"journal":{"name":"Zebrafish","volume":null,"pages":null},"PeriodicalIF":2.0,"publicationDate":"2022-04-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45608156","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cytogenetic and Molecular Characterization of Oligosarcus pintoi (Characidae): A New Record of Supernumerary Chromosome in this Species.","authors":"Isabelle Pereira Mari-Ribeiro, Bárbara Scorsim, A. V. D. Oliveira, A. L. Portela-Castro","doi":"10.1089/zeb.2021.0065","DOIUrl":"https://doi.org/10.1089/zeb.2021.0065","url":null,"abstract":"The genus Oligosarcus currently comprises 24 valid species distributed in the major river basins of South America. In this group, nine species were cytogenetically investigated, and found to share a diploid number of 50 chromosomes. Despite the conservation of the diploid number, variations in the karyotypic formula, number and position of the nucleolar organizer regions, and longitudinal bands have been described between both species and populations. In this study, we present cytogenetic and molecular data from Oligosarcus pintoi specimens from the Keller River, a tributary of the Ivaí River (Upper Paraná basin), using DNA barcoding and cytogenetic markers (C-band, silver-stained nucleolar organizer regions, and fluorescence in situ hybridization of 18S and 5S rDNA). The genetic inferences reached after analyzing the cytochrome c oxidade subunit 1 gene allowed us to confirm the identity of the individuals with 2n = 50 chromosomes. However, one specimen contained a medium subtelocentric supernumerary chromosome (2n = 51). This is the second record of additional chromosomes in O. pintoi, thereby confirming the existence of a supernumerary chromosome in allopatric populations of this species, a fact that demonstrates an evolutionary path that is divergent from other populations and/or species of Oligosarcus analyzed so far, contributing to the karyotypic diversification of the group.","PeriodicalId":23872,"journal":{"name":"Zebrafish","volume":null,"pages":null},"PeriodicalIF":2.0,"publicationDate":"2022-04-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45829349","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}