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Heritable CRISPR Mutagenesis of Essential Maternal Effect Genes as a Simple Tool for Sustained Population Suppression of Invasive Species in a Zebrafish Model. 在斑马鱼模型中用可遗传的 CRISPR 基因突变法对重要的母体效应基因进行突变，以此作为持续抑制入侵物种种群的简单工具。
Zebrafish Pub Date : 2024-08-01 Epub Date: 2024-03-20 DOI: 10.1089/zeb.2023.0108
Christopher J Krueger, Zhifei Dai, Cheng Zhu, Bo Zhang
{"title":"Heritable CRISPR Mutagenesis of Essential Maternal Effect Genes as a Simple Tool for Sustained Population Suppression of Invasive Species in a Zebrafish Model.","authors":"Christopher J Krueger, Zhifei Dai, Cheng Zhu, Bo Zhang","doi":"10.1089/zeb.2023.0108","DOIUrl":"10.1089/zeb.2023.0108","url":null,"abstract":"<p><p>Invasive species control is important for ecological and agricultural management. Genetic methods can provide species specificity for population control. We developed heritable maternal effect embryo lethality (HMEL), a novel strategy allowing negative population pressure from HMEL individuals to be transmitted within a population across generations. We demonstrate the HMEL technique in zebrafish through genome-integrated CRISPR/Cas targeted mutagenic disruption of <i>nucleoplasmin 2b (npm2b)</i>, a female-specific essential maternal effect gene, causing heritable sex-limited disruption of reproduction. <i>HMEL</i>-induced high-efficiency mutation of <i>npm2b</i> in females suppresses population, while males transmit the <i>HMEL</i> allele across generations. HMEL could be easily modified to target other genes causing sex-specific sterility, or generalized to control invasive fish or other vertebrate species for environmental conservation or agricultural protection.</p>","PeriodicalId":94273,"journal":{"name":"Zebrafish","volume":" ","pages":"279-286"},"PeriodicalIF":0.0,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140178530","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
Integrating Genetic and Cytogenetic Data: A Diversity Study of Astyanax and Psalidodon (Characidae) Species from the Paraná River Basin. 整合遗传学和细胞遗传学数据：巴拉那河流域 Astyanax 和 Psalidodon (Characidae) 鱼种的多样性研究。
Zebrafish Pub Date : 2024-08-01 Epub Date: 2024-05-30 DOI: 10.1089/zeb.2024.0134
Isabelle Pereira Mari-Ribeiro, Bárbara Scorsim, Leandro Ranucci, Luciana Andréia Borin-Carvalho, Alessandra Valéria de Oliveira, Ana Luiza de Brito Portela-Castro
{"title":"Integrating Genetic and Cytogenetic Data: A Diversity Study of <i>Astyanax</i> and <i>Psalidodon</i> (Characidae) Species from the Paraná River Basin.","authors":"Isabelle Pereira Mari-Ribeiro, Bárbara Scorsim, Leandro Ranucci, Luciana Andréia Borin-Carvalho, Alessandra Valéria de Oliveira, Ana Luiza de Brito Portela-Castro","doi":"10.1089/zeb.2024.0134","DOIUrl":"10.1089/zeb.2024.0134","url":null,"abstract":"<p><p><i>Astyanax</i> is one of the most specious fish groups in the Neotropical region, with many cryptic species, which represents a challenge for correct identification through traditional taxonomic methods. <i>Psalidodon</i> is a recently resurrected genus group of species previously belonging to <i>Astyanax</i>, specifically those with extensive chromosomal variation of the <i>A. scabripinnis</i> and <i>fasciatu</i>s complexes. In the present study, the mitochondrial genes cytochrome <i>c</i> oxidase subunit 1 (COI), mitochondrial ATP synthase 6 and 8 (ATPase 6/8), and NADH dehydrogenase subunit 2 (ND2) were used in conjunction with chromosomal data to characterize molecularly and cytogenetically populations of <i>Astyanax</i> and <i>Psalidodon</i> from rivers and streams of the Ivaí River Basin (Paraná Basin). The results demonstrated the effectiveness of the integrative use of molecular and cytogenetic techniques, with the confirmation of at least three species for the sampled sites: <i>A. lacustris</i>, <i>P. paranae</i>, and <i>P. fasciatus</i>, which showed inter- and intrapopulation karyotype variations. In addition, extensive haplotypic variation can be observed for these species within the Ivaí River Basin and throughout the Paraná River Basin. The data demonstrate a hidden diversity among the species analyzed, enrich the ichthyofaunistic knowledge of small rivers and streams, and contribute to future conservation projects in these areas.</p>","PeriodicalId":94273,"journal":{"name":"Zebrafish","volume":" ","pages":"300-309"},"PeriodicalIF":0.0,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141176902","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
Genotyping Zebrafish Point Mutant by Allele-Specific Blocking PCR. 通过等位基因特异性阻断 PCR 对斑马鱼点突变体进行基因分型。
Zebrafish Pub Date : 2024-08-01 Epub Date: 2024-05-29 DOI: 10.1089/zeb.2024.0138
Lih Khiang Beh, Hongyuan Shen
{"title":"Genotyping Zebrafish Point Mutant by Allele-Specific Blocking PCR.","authors":"Lih Khiang Beh, Hongyuan Shen","doi":"10.1089/zeb.2024.0138","DOIUrl":"10.1089/zeb.2024.0138","url":null,"abstract":"<p><p>Genotyping zebrafish carrying wild-type, heterozygous, or homozygous copies of a mutant allele is often required for investigating gene specific functions, and is routinely performed to differentiate point mutants. In this study, we describe a modified allele-specific PCR method using an additional blocking primer to promote target sequence amplification while suppressing sequences with single mismatch. Using the <i>tp53<sup>m214k</sup></i> point mutant as an example, we show that wild-type, heterozygous, and homozygous zebrafish can be easily distinguished using this simple PCR method, which could be widely adapted for genotyping zebrafish with point mutations or small nucleotide insertions/deletions.</p>","PeriodicalId":94273,"journal":{"name":"Zebrafish","volume":" ","pages":"297-299"},"PeriodicalIF":0.0,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141162970","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
Zebrafish from the Alps to Sicily: The 4th Italian Zebrafish Meeting in Palermo. 从阿尔卑斯山到西西里岛的斑马鱼：在巴勒莫举行的第四届意大利斑马鱼会议。
Zebrafish Pub Date : 2024-08-01 Epub Date: 2024-07-04 DOI: 10.1089/zeb.2024.0140
Massimiliano Andreazzoli, Francesco Argenton, Cristiano Bertolucci, Dario Finazzi, Chiara Gabellini, Marina Mione, Anna Pistocchi, Massimo Santoro, Paolo Sordino, Vincenzo Cavalieri
{"title":"Zebrafish from the Alps to Sicily: The 4th Italian Zebrafish Meeting in Palermo.","authors":"Massimiliano Andreazzoli, Francesco Argenton, Cristiano Bertolucci, Dario Finazzi, Chiara Gabellini, Marina Mione, Anna Pistocchi, Massimo Santoro, Paolo Sordino, Vincenzo Cavalieri","doi":"10.1089/zeb.2024.0140","DOIUrl":"10.1089/zeb.2024.0140","url":null,"abstract":"<p><p>The 4th Italian Zebrafish Meeting took place in Palermo from February 7 to 9, 2024. The primary aim of this meeting was to bring together a diverse group of principal investigators, young researchers, facility managers, commercial vendors, and others to provide an important forum for presentation and discussion of the most innovative and exciting scientific research currently ongoing in Italy using the zebrafish model. Nonetheless, the meeting program has been conceived to allow the dissemination of cutting-edge scientific research across a wide range of topics and to shed light on its future directions, without geographical boundaries. Indeed, people from various parts of the world joined the meeting, and 210 participants presented their latest work in talks and posters. Importantly, the meeting had designated time to foster open scientific exchange and informal networking opportunities among participants of all career stages, thus allowing initiation of new collaborations and strengthening of existing partnerships. The meeting was a tremendous success as testified by the highest participation ever since the first meeting of the series in 2017, coupled with the highly positive satisfaction rating expressed by the attendants. The full program and detailed information about the meeting can be found on the dedicated website at https://itazebrafishmeeting.wixsite.com/izm2024.</p>","PeriodicalId":94273,"journal":{"name":"Zebrafish","volume":" ","pages":"275-278"},"PeriodicalIF":0.0,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141500111","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
Fish Playpens: Method for Raising Individual Juvenile Zebrafish on a Recirculating System for Studies Requiring Repeated Measures. 鱼类游戏池：在需要重复测量的研究中使用循环系统饲养斑马鱼幼鱼的方法。
Zebrafish Pub Date : 2024-08-01 Epub Date: 2023-12-28 DOI: 10.1089/zeb.2023.0103
Tabea O C Moll, Steven A Farber
{"title":"Fish Playpens: Method for Raising Individual Juvenile Zebrafish on a Recirculating System for Studies Requiring Repeated Measures.","authors":"Tabea O C Moll, Steven A Farber","doi":"10.1089/zeb.2023.0103","DOIUrl":"10.1089/zeb.2023.0103","url":null,"abstract":"<p><p>Even though many experimental approaches benefit from tracking individual juvenile animals, there is yet to be a commercial zebrafish rack system designed to accomplish this task. Thus, we invented playpens, an acrylic, and screen container, to raise 12 individual zebrafish juveniles per standard 10 L tank on an existing recirculating fish system. During a week-long experiment, fish raised in playpens grow to the same size as conventionally raised juveniles.</p>","PeriodicalId":94273,"journal":{"name":"Zebrafish","volume":" ","pages":"294-296"},"PeriodicalIF":0.0,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139050038","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 Robust Closed-Tube Method for Resolving tp53M214K Genotypes. 解决 tp53M214K 基因型问题的可靠闭管方法
Zebrafish Pub Date : 2024-06-01 Epub Date: 2024-02-20 DOI: 10.1089/zeb.2023.0030
Katherine M Silvius, Genevieve C Kendall
{"title":"A Robust Closed-Tube Method for Resolving <i>tp53</i><sup>M214K</sup> Genotypes.","authors":"Katherine M Silvius, Genevieve C Kendall","doi":"10.1089/zeb.2023.0030","DOIUrl":"10.1089/zeb.2023.0030","url":null,"abstract":"<p><p>The <i>tp53</i><sup>M214K</sup> zebrafish mutant is a versatile platform with which to model a diverse spectrum of human diseases. However, currently available genotyping methods for this mutant require lengthy hands-on processes such as restriction digests and outsourced Sanger sequencing. To address this deficiency, we leveraged high-resolution melting analysis technology in conjunction with a parallel, in-tandem wild-type spike-in approach to develop a robust genotyping protocol capable of discriminating <i>tp53</i><sup>M214K</sup> zygosity. In this study, we describe our method in detail. We anticipate that our genotyping protocol will benefit researchers utilizing the <i>tp53</i><sup>M214K</sup> zebrafish mutant by offering reliable results with a shorter turnaround time, lower personnel involvement, and higher throughput than traditional methods, thereby decreasing the burden of genotyping and maximizing research efficiency.</p>","PeriodicalId":94273,"journal":{"name":"Zebrafish","volume":" ","pages":"250-254"},"PeriodicalIF":0.0,"publicationDate":"2024-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11296207/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139935016","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
Initial Steps of XY Sex Chromosome Differentiation in the Armored Catfish Hypostomus albopunctatus (Siluriformes: Loricariidae) Revealed by Heterochromatin Accumulation. 异染色质的积累揭示了铠甲鲶（Siluriformes: Loricariidae）XY 性染色体分化的初始步骤
Zebrafish Pub Date : 2024-06-01 Epub Date: 2024-02-20 DOI: 10.1089/zeb.2023.0100
Ligia Carla Balini, Carlos Alexandre Fernandes, Ana Luiza de Brito Portela-Castro, Rafael Fernando de Melo, Cláudio Henrique Zawadzki, Luciana Andreia Borin-Carvalho
{"title":"Initial Steps of XY Sex Chromosome Differentiation in the Armored Catfish <i>Hypostomus albopunctatus</i> (Siluriformes: Loricariidae) Revealed by Heterochromatin Accumulation.","authors":"Ligia Carla Balini, Carlos Alexandre Fernandes, Ana Luiza de Brito Portela-Castro, Rafael Fernando de Melo, Cláudio Henrique Zawadzki, Luciana Andreia Borin-Carvalho","doi":"10.1089/zeb.2023.0100","DOIUrl":"10.1089/zeb.2023.0100","url":null,"abstract":"<p><p>In fish species, heterochromatinization is one process that could trigger sex chromosome differentiation. The present article describes a nascent XX/XY sex chromosome system evidenced by heterochromatin accumulation and microsatellite (GATA)<sub>8</sub> in <i>Hypostomus albopunctatus</i> from two populations of the Paraná River basin. The specimens of <i>H. albopunctatus</i> from the Campo and Bossi Rivers share the same karyotype. The species exhibits 74 chromosomes (8m+14sm +16st +36a, fundamental number = 112). The C-banding technique suggests male heterogamety in <i>H. albopunctatus</i>, where the Y-chromosome is morphologically like the X-chromosome but differs from it for having long arms that are entirely heterochromatic. Double fluorescence <i>in situ</i> hybridization (FISH) with 18S and 5S rDNA probes confirmed the Ag-nucleolus organizer region sites in a single pair for both populations, and minor rDNA clusters showed interpopulational variation. FISH with the microsatellite (GATA)<sub>8</sub> probe showed a dispersed pattern in the karyotype, accumulating these sequences of sex chromosomes of both populations. FISH with microsatellite (CGC)<sub>10</sub> probe showed interpopulational variation. The absence of differentiated sex chromosomes in <i>H. albopunctatus</i> is described previously, and a new variant is documented herein where XY chromosomes can be seen in an early stage of differentiation.</p>","PeriodicalId":94273,"journal":{"name":"Zebrafish","volume":" ","pages":"265-273"},"PeriodicalIF":0.0,"publicationDate":"2024-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139935017","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
Defining the Unseen: Population-Specific Markers for Astyanax mexicanus Blind Cavefish. 定义看不见的东西：墨西哥盲洞鱼种群特异性标记。
Zebrafish Pub Date : 2024-06-01 Epub Date: 2024-02-14 DOI: 10.1089/zeb.2023.0105
Aubrey E Manning, Hannah Grunwald, Rachel Moran, Roberto Rodriguez-Morales, Amanda K Powers, Suzanne McGaugh, Johanna E Kowalko
{"title":"Defining the Unseen: Population-Specific Markers for <i>Astyanax mexicanus</i> Blind Cavefish.","authors":"Aubrey E Manning, Hannah Grunwald, Rachel Moran, Roberto Rodriguez-Morales, Amanda K Powers, Suzanne McGaugh, Johanna E Kowalko","doi":"10.1089/zeb.2023.0105","DOIUrl":"10.1089/zeb.2023.0105","url":null,"abstract":"<p><p><i>Astyanax mexicanus</i> is an emerging model system used to study development, evolution, and behavior of multiple cavefish populations that have repeatedly evolved from conspecific surface fish. Although surface and cavefish live and breed in the laboratory, there are no rapid methods for distinguishing between different cavefish populations. We present 2 methods for genotyping fish for a total of 16 population-specific markers using methods that are easy and inexpensive to implement in a basic molecular biology laboratory. This resource will help researchers maintain independent stocks within the laboratory and distinguish between fish from different populations.</p>","PeriodicalId":94273,"journal":{"name":"Zebrafish","volume":" ","pages":"255-258"},"PeriodicalIF":0.0,"publicationDate":"2024-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11301698/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139737046","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 Chemically Inducible Muscle Ablation System for the Zebrafish. 斑马鱼化学诱导肌肉消融系统
Zebrafish Pub Date : 2024-06-01 Epub Date: 2024-03-04 DOI: 10.1089/zeb.2023.0102
Eric Paulissen, Benjamin L Martin
{"title":"A Chemically Inducible Muscle Ablation System for the Zebrafish.","authors":"Eric Paulissen, Benjamin L Martin","doi":"10.1089/zeb.2023.0102","DOIUrl":"10.1089/zeb.2023.0102","url":null,"abstract":"<p><p>An effective method for tissue-specific ablation in zebrafish is the nitroreductase (NTR)/metronidazole (MTZ) system. Expressing bacterial NTR in the presence of nitroimidazole compounds causes apoptotic cell death, which can be useful for understanding many biological processes. However, this requires tissue-specific expression of the NTR enzyme, and many tissues have yet to be targeted with transgenic lines that express NTR. We generated a transgenic zebrafish line expressing NTR in differentiated skeletal muscle. Treatment of embryos with MTZ caused muscle specific cell ablation. We demonstrate this line can be used to monitor muscle regeneration in whole embryos and in transplanted transgenic cells.</p>","PeriodicalId":94273,"journal":{"name":"Zebrafish","volume":" ","pages":"243-249"},"PeriodicalIF":0.0,"publicationDate":"2024-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11301710/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140023896","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
Modified Procedure for Simple and Rapid Collection of Zebrafish Fluid Samples for Biochemical Analyses. 简单快速采集斑马鱼体液样本进行生化分析的改进程序。
Zebrafish Pub Date : 2024-06-01 Epub Date: 2024-02-20 DOI: 10.1089/zeb.2023.0101
Ana Paula Apolinário da Silva, João Xavier da Silva Neto, José Ytalo Gomes da Silva, Ana Lorena Pereira Bezerra, Bruno Bezerra da Silva, Maria Izabel Florindo Guedes
{"title":"Modified Procedure for Simple and Rapid Collection of Zebrafish Fluid Samples for Biochemical Analyses.","authors":"Ana Paula Apolinário da Silva, João Xavier da Silva Neto, José Ytalo Gomes da Silva, Ana Lorena Pereira Bezerra, Bruno Bezerra da Silva, Maria Izabel Florindo Guedes","doi":"10.1089/zeb.2023.0101","DOIUrl":"10.1089/zeb.2023.0101","url":null,"abstract":"<p><p>This study addresses the challenge of collecting blood samples from zebrafish for biochemical analysis. Traditional methods are cumbersome due to low blood flow and rapid coagulation. Based on a previously published technique, we simplified the process by applying an anticoagulant solution directly to the incision site. The modified protocol involves immersing the fish in an ice bath, making a cross-sectional incision, and immediately applying anticoagulant solution. Centrifugation of the specimens provides a streamlined and efficient approach to zebrafish fluid sample collection, compatible with classic biochemical marker analyses.</p>","PeriodicalId":94273,"journal":{"name":"Zebrafish","volume":" ","pages":"259-264"},"PeriodicalIF":0.0,"publicationDate":"2024-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139935018","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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