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Novel Husbandry Practices Result in Rapid Rates of Growth and Sexual Maturation Without Impacting Adult Behavior in the Blind Mexican Cavefish. 新的饲养方法可使墨西哥洞穴盲鱼快速生长和性成熟,且不会影响其成年行为。
IF 2 4区 生物学 Q1 Agricultural and Biological Sciences Pub Date : 2023-04-01 DOI: 10.1089/zeb.2023.0001
Robert A Kozol, Anders Yuiska, Ji Heon Han, Bernadeth Tolentino, Arthur Lopatto, Peter Lewis, Alexandra Paz, Alex C Keene, Johanna E Kowalko, Erik R Duboué

Animal model systems are dependent on the standardization of husbandry protocols that maximize growth and reduce generation time. The Mexican tetra, Astyanax mexicanus, exists as eyed surface and blind cave dwelling populations. The opportunity for comparative approaches between independently evolved populations has led to the rapid growth of A. mexicanus as a model for evolution and biomedical research. However, a slow and inconsistent growth rate remains a major limitation to the expanded application of A. mexicanus. Fortunately, this temporal limitation can be addressed through husbandry changes that accelerate growth rates while maintaining optimal health outcomes. Here, we describe a husbandry protocol that produces rapid growth rates through changes in diet, feeding frequency, growth sorting and progressive changes in tank size. This protocol produced robust growth rates and decreased the age of sexual maturity in comparison to our previous protocol. To determine whether changes in feeding impacted behavior, we tested fish in exploration and schooling assays. We found no difference in behavior between the two groups, suggesting that increased feeding and rapid growth will not impact the natural variation in behavioral traits. Taken together, this standardized husbandry protocol will accelerate the development of A. mexicanus as a genetic model.

动物模型系统有赖于标准化的饲养规程,以最大限度地提高生长速度并缩短生成时间。墨西哥四带喙四带喙四带喙四带喙四带喙四带喙四带喙四带喙四带喙四带喙四带喙四带喙四带喙四带喙由于有机会在独立进化的种群之间进行比较,墨西哥四眼蛙迅速成长为进化和生物医学研究的典范。然而,缓慢和不一致的生长速度仍然是扩大墨西哥猿猴应用范围的主要限制因素。幸运的是,这种时间限制可以通过改变饲养方式来解决,从而在保持最佳健康结果的同时加快生长速度。在此,我们介绍了一种饲养方案,该方案通过改变饮食、喂食频率、生长分类和逐步改变水箱大小来提高生长速度。与我们之前的方案相比,该方案不仅能提高生长速度,还能降低性成熟年龄。为了确定投喂量的变化是否会影响鱼的行为,我们对鱼进行了探索和游弋试验。我们发现两组鱼的行为没有差异,这表明增加喂食和快速生长不会影响行为特征的自然变化。综上所述,这种标准化饲养方法将加速墨西哥鲶鱼作为遗传模型的发展。
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引用次数: 0
Acknowledgment of Reviewers 2022. 审稿人致谢2022。
IF 2 4区 生物学 Q1 Agricultural and Biological Sciences Pub Date : 2023-02-01 DOI: 10.1089/zeb.2022.29008.ack
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引用次数: 0
Zebrafish sox2 Is Required for the Swim Bladder Inflation by Controlling the Swim-Up Behavior. 斑马鱼需要sox2通过控制游动行为来调节鱼鳔膨胀。
IF 2 4区 生物学 Q1 Agricultural and Biological Sciences Pub Date : 2023-02-01 DOI: 10.1089/zeb.2022.0043
Shasha Cao, Zhangji Dong, Xiaohua Dong, Wenshuang Jia, Fuyou Zhou, Qingshun Zhao

The swim bladder functions to maintain the fish balance at a certain position under water. Although the motoneuron-dependent swim-up behavior is important for swim bladder inflation, the underlying molecular mechanism remains largely unknown. We generated a sox2 KO zebrafish using TALEN and found that the posterior chamber of the swim bladder was uninflated. The tail flick and the swim-up behavior were absent in the mutant zebrafish embryos and the behavior could not be accomplished. As the tail flick behavior is absent, the mutant larvae therefore cannot reach the water surface to gulp air, ultimately leading to the uninflation of the swim bladder. To understand the mechanism underlying the swim-up defects, we crossed the sox2 null allele in the background of Tg(huc:eGFP) and Tg(hb9:GFP). The deficiency of sox2 in zebrafish resulted in abnormal motoneuron axons in the regions of trunk, tail, and swim bladder. To identify the downstream target gene of sox2 to control the motor neuron development, we performed RNA sequencing on the transcriber of mutant embryos versus wild type embryos and found that the axon guidance pathway was abnormal in the mutant embryos. RT-PCR demonstrated that the expression of sema3bl, ntn1b, and robo2 were decreased in the mutants.

鱼鳔的作用是维持鱼在水下某一位置的平衡。尽管运动神经元依赖的上浮行为对鱼鳔膨胀很重要,但其潜在的分子机制仍不清楚。我们用TALEN生成了一条sox2 KO斑马鱼,发现鱼鳔的后腔没有充气。突变的斑马鱼胚胎中没有甩尾和上浮行为,无法完成这一行为。由于没有甩尾行为,突变幼虫因此无法到达水面吞咽空气,最终导致鱼鳔不膨胀。为了了解游动缺陷的机制,我们在Tg(huc:eGFP)和Tg(hb9:GFP)的背景下交叉了sox2空等位基因。斑马鱼缺乏sox2导致躯干、尾部和鱼鳔区域的运动神经元轴突异常。为了确定sox2调控运动神经元发育的下游靶基因,我们对突变胚胎与野生型胚胎的转录子进行了RNA测序,发现突变胚胎的轴突引导通路异常。RT-PCR结果显示,突变体中sema3bl、ntn1b和robo2的表达减少。
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引用次数: 0
Chromosomal Mapping of the Histone Multigene Family and U2 snRNA in Hypancistrus Species (Siluriformes, Loricariidae) from the Brazilian Amazon. 巴西亚马逊河流域大腹蛇(Siluriformes, Loricariidae)组蛋白多基因家族和U2 snRNA染色体定位
IF 2 4区 生物学 Q1 Agricultural and Biological Sciences Pub Date : 2023-02-01 DOI: 10.1089/zeb.2022.0030
Carlos Eduardo Vasconcelos Dos Santos, Bruno Rafael Ribeiro de Almeida, Flávia Dos Santos Tavares, Luan Felipe da Silva Frade, Adauto Lima Cardoso, André Luiz Alves de Sá, Cleusa Yoshiko Nagamachi, Leandro Melo de Sousa, Julio Cesar Pieczarka, Renata Coelho Rodrigues Noronha

Loricariidae (Siluriformes) comprises ∼1026 species of neotropical fish, being considered the most diverse among the Siluriformes. Studies on repetitive DNA sequences have provided important data on the evolution of the genomes of members of this family, especially of the Hypostominae subfamily. In this study, the chromosomal mapping of the histone multigene family and U2 snRNA was performed in two species belonging to the Hypancistrus genus, Hypancistrus sp. "pão" (2n = 52, 22m + 18sm +12st) and Hypancistrus zebra (2n = 52, 16m + 20sm +16st). The presence of dispersed signals of histones H2A, H2B, H3, and H4 in the karyotype of both species, with each sequence displaying a varied level of accumulation and dispersion of these sequences between them was observed; in addition, U2 snDNA probe only showed positive results in H. zebra, which present this multigene in the terminal region of three chromosomal pairs. The obtained results resemble data already analyzed in the literature, in which the action of transposable elements interfere in the organization of these multigene families, in addition to other evolutionary processes that shape the evolution of the genome, such as circular or ectopic recombination. This study also shows that the dispersion of the multigene histone family is quite complex, and from this, these data serve as a point of discussion for the evolutionary processes that occur in the Hypancistrus karyotype.

Loricariidae (Siluriformes)包括约1026种新热带鱼,被认为是Siluriformes中最多样化的。重复DNA序列的研究为该科成员的基因组进化提供了重要的数据,特别是下stominae亚科。本研究对Hypancistrus属(Hypancistrus sp.)的两个物种进行了组蛋白多基因家族和U2 snRNA的染色体定位。“p o”(2n = 52,22m + 18sm +12st)和Hypancistrus zebra (2n = 52,16m + 20sm +16st)。组蛋白H2A、H2B、H3和H4在两种物种核型中均存在分散信号,且各序列在两种物种之间表现出不同程度的积累和分散;此外,U2 snDNA探针仅在斑马斑马中显示阳性结果,该多基因存在于三对染色体的末端区。所获得的结果与文献中已经分析的数据相似,其中转座因子的作用干扰了这些多基因家族的组织,以及其他形成基因组进化的进化过程,例如圆形或异位重组。本研究还表明,多基因组蛋白家族的分散是相当复杂的,从这一点来看,这些数据可以作为Hypancistrus核型中发生的进化过程的讨论点。
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引用次数: 1
Molecular Markers Reveal a New and Possibly Threatened Species of Cnesterodon (Poeciliidae, Cnesterodontini) from the Upper Paraná River Basin, Brazil. 分子标记揭示了巴西巴拉那河上游流域一种可能濒临灭绝的 Cnesterodon(Poeciliidae,Cnesterodontini)新物种。
IF 2 4区 生物学 Q1 Agricultural and Biological Sciences Pub Date : 2023-02-01 Epub Date: 2023-01-25 DOI: 10.1089/zeb.2022.0052
João Paulo Morais-Silva, Bárbara Scorsim, Geovana Gonçalves, Augusto Frota, Weferson Júnio da Graça, Alessandra Valéria de Oliveira

Cnesterodon comprises 10 valid species occurring in the major river basins of South America. Recent ichthyofaunistic studies in the Ivaí River basin, upper Paraná River system, suggested the existence of a possible new species, which was identified as Cnesterodon sp. based on morphological characters. Currently, the use of molecular tools has proved to be fundamental in aiding phylogenetics and cataloging biodiversity; therefore, in this study, we molecularly characterize a possible new species of Cnesterodon from the Ivaí River basin encoding the mitochondrial genes Cytochrome c Oxidase, subunit I (COI), and NADH dehydrogenase subunit 2 (ND2). The genetic differences found showed that this species really differs from the other Cnesterodon species, indicating that it is a distinct species, which is possibly already in serious danger of extinction since its habitat often suffers from human exploitation and its distribution is restricted to only two sites in the upper Ivaí River basin, but it has disappeared in one of them.

Cnesterodon包括10个有效物种,分布于南美洲的主要河流流域。最近在巴拉那河上游伊瓦伊河流域进行的鱼类学研究表明可能存在一个新物种,根据形态特征将其鉴定为 Cnesterodon sp.。目前,分子工具的使用已被证明是辅助系统发生学和生物多样性编目的基础;因此,在本研究中,我们通过编码线粒体基因细胞色素 c 氧化酶亚基 I(COI)和 NADH 脱氢酶亚基 2(ND2),对伊瓦伊河流域可能存在的 Cnesterodon 新物种进行了分子鉴定。发现的遗传差异表明,该物种确实不同于其他 Cnesterodon 物种,表明它是一个独特的物种,可能已经面临灭绝的严重危险,因为其栖息地经常遭受人类开发,其分布仅限于伊瓦伊河流域上游的两个地点,但在其中一个地点已经消失。
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引用次数: 0
Metoclopramide as a Potential Antipsychotic Against Long-Term Methionine Exposure in Zebrafish. 甲氧氯普胺对斑马鱼长期蛋氨酸暴露的潜在抗精神病药物作用。
IF 2 4区 生物学 Q1 Agricultural and Biological Sciences Pub Date : 2023-02-01 DOI: 10.1089/zeb.2022.0033
Hemen Ved, Gaurav Doshi, Nirav Bhatia, Pravin Kale

Methionine (MET) contributes to brain function and is required for proper functioning of the central nervous system. However, exceptionally high levels of MET and its metabolites in plasma have been found to be toxic and can lead to cell alterations. Long-term exposure to MET has been shown to mimic psychotic symptoms in schizophrenic patients and rodents. The present study evaluated behavioral and neurochemical effects of long-term exposure to MET in zebrafish. Five groups of zebrafish were exposed to MET at a concentration of 4.5 mM for 7 days, along with acute exposure to 25 μM of clozapine and 750, 1000, and 1250 μM of metoclopramide. In contrast, the normal group was exposed to only water and dimethyl sulfoxide. After the treatment, social interaction, anxiety, memory, and locomotion of zebrafish and serotonin levels in zebrafish brains were evaluated. Our results showed that metoclopramide was not only beneficial in improving MET-induced cognitive impairment but it also prevented social withdrawal in zebrafish exposed to MET. In addition, metoclopramide reversed anxiety-like behavior, as indicated by significant changes in locomotion activity. Despite slight changes in serotonin levels in the zebrafish brain, an in vitro serotonin assay failed to demonstrate significant differences between the disease control, normal, and two treatment groups. Finally, results from the study showed that repeated administration of MET induced schizophrenia-like symptoms, although metoclopramide ameliorated the MET-mediated negative symptoms and cognitive deficits in zebrafish. Overall, our findings suggest a new perspective to further explore the antipsychotic properties of metoclopramide.

蛋氨酸(MET)有助于大脑功能,是中枢神经系统正常运作所必需的。然而,血浆中异常高水平的MET及其代谢物已被发现是有毒的,并可能导致细胞改变。长期暴露于MET已被证明可以模仿精神分裂症患者和啮齿动物的精神病症状。本研究评估了斑马鱼长期暴露于MET对行为和神经化学的影响。5组斑马鱼连续7天暴露于浓度为4.5 mM的MET,同时急性暴露于25 μM的氯氮平和750、1000和1250 μM的甲氧氯普胺。相比之下,正常组只暴露于水和二甲亚砜。治疗后,评估斑马鱼的社会互动、焦虑、记忆和运动以及斑马鱼大脑中的血清素水平。我们的研究结果表明,甲氧氯普胺不仅有利于改善MET诱导的认知障碍,而且还可以防止暴露于MET的斑马鱼的社交退缩。此外,甲氧氯普胺逆转焦虑样行为,运动活动的显著变化表明。尽管斑马鱼大脑中的血清素水平略有变化,但体外血清素测定未能证明疾病控制组、正常组和两个治疗组之间存在显著差异。最后,研究结果表明,尽管甲氧氯普胺改善了斑马鱼中MET介导的阴性症状和认知缺陷,但反复给予MET诱导精神分裂症样症状。总之,我们的发现为进一步探索甲氧氯普胺的抗精神病特性提供了一个新的视角。
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引用次数: 1
wnt11f2 Zebrafish, an Animal Model for Development and New Insights in Bone Formation. wnt11f2斑马鱼:发育的动物模型和骨形成的新见解。
IF 2 4区 生物学 Q1 Agricultural and Biological Sciences Pub Date : 2023-02-01 DOI: 10.1089/zeb.2022.0042
Caroline Caetano da Silva, Agnes Ostertag, Ratish Raman, Marc Muller, Martine Cohen-Solal, Corinne Collet

Wnt signaling is a key regulator of osteoblast differentiation and mineralization in humans and animals, mediated by the canonical Wnt/β-catenin and noncanonical signaling pathways. Both pathways are crucial in regulating osteoblastogenesis and bone formation. The zebrafish silberblick (slb) carries a mutation in wnt11f2, a gene that contributes to embryonic morphogenesis; however, its role in bone morphology is unknown. wnt11f2 was originally known as wnt11; it was recently reclassified to avoid confusion in comparative genetics and disease modeling. The goal of this review is to summarize the characterization of the wnt11f2 zebrafish mutant and to deliver some new insights concerning its role in skeletal development. In addition to the previously described defects in early development in this mutant as well as craniofacial dysmorphia, we show an increase in tissue mineral density in the heterozygous mutant that points to a possible role of wnt11f2 in high bone mass phenotypes.

Wnt信号是人类和动物成骨细胞分化和矿化的关键调节因子,由典型的Wnt/β-catenin和非典型的信号通路介导。这两种途径在调节成骨细胞发生和骨形成中都是至关重要的。斑马鱼silberblick (slb)携带wnt11f2基因突变,该基因有助于胚胎形态发生;然而,其在骨形态中的作用尚不清楚。Wnt11f2最初被称为wnt11;它最近被重新分类,以避免在比较遗传学和疾病模型中混淆。本文的目的是总结wnt11f2斑马鱼突变体的特征,并就其在骨骼发育中的作用提供一些新的见解。除了先前描述的该突变体早期发育缺陷以及颅面畸形外,我们还发现杂合突变体的组织矿物质密度增加,这表明wnt11f2可能在高骨量表型中起作用。
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引用次数: 0
Call for Special Issue Papers: 2023 Education Issue of Zebrafish. 征稿:2023年斑马鱼教育特刊。
IF 2 4区 生物学 Q1 Agricultural and Biological Sciences Pub Date : 2022-12-01 DOI: 10.1089/zeb.2022.29009.cfp
Lara Hutson, Jennifer Liang, Michael Pickart, Christopher Pierret, Tamara Franz-Odendaal, Madelyn Jackstadt
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引用次数: 0
Common Aquarium Plants as an Enrichment Strategy in Zebrafish Facilities. 常见水族植物在斑马鱼设施中的富集策略。
IF 2 4区 生物学 Q1 Agricultural and Biological Sciences Pub Date : 2022-12-01 DOI: 10.1089/zeb.2022.0036
Benjamin Tsang, Robert T Gerlai

An increasing number of laboratories utilize zebrafish as this species is now represented in practically every subfield of biology research. Environmental enrichment has been shown to improve welfare and health of a large number of species of animals studied in laboratories, kept in zoos, or used in agriculture. However, most laboratories keep zebrafish in barren tanks. Artificial and live aquatic plants have been used in a variety of contexts for aquarium fish, and they have also been suggested as a potential enrichment strategy for the zebrafish. However, no systematic studies have been conducted to investigate their beneficial effects in zebrafish husbandry and biology research. In this study, we review some of the potential benefits of using live plants, and argue that systematic analyses for proper choice and use of live plants in zebrafish husbandry are sorely needed. We provide a few examples of aquatic plant species out of the large variety produced for the aquarium hobby that could be adopted to the zebrafish laboratory setting as environmental enrichment. We speculate that once systematic analyses have been conducted, they will show benefits of this ethologically/ecologically relevant enrichment method, one of which will be healthier and less stressed fish leading to increased reproducibility of results in zebrafish research.

越来越多的实验室利用斑马鱼,因为这个物种现在在生物学研究的几乎每个子领域都有代表。环境富集已被证明可以改善实验室、动物园或农业中使用的大量动物的福利和健康。然而,大多数实验室将斑马鱼饲养在贫瘠的水箱中。人工和活的水生植物已经在各种情况下用于观赏鱼,它们也被认为是斑马鱼的潜在富集策略。然而,目前还没有对其在斑马鱼养殖和生物学研究中的有益作用进行系统的研究。在本研究中,我们回顾了利用活植物的一些潜在好处,并认为有必要对活植物在斑马鱼养殖中的正确选择和使用进行系统分析。我们提供了一些水生植物物种的例子,这些水生植物物种是为水族馆的爱好而生产的,可以采用斑马鱼实验室设置作为环境富集。我们推测,一旦进行了系统的分析,它们将显示出这种与行为学/生态学相关的富集方法的好处,其中一种富集方法将更健康,压力更小,从而提高斑马鱼研究结果的可重复性。
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引用次数: 1
Comparative Analysis of Central Corneal Thickness in Four Fish Models. 四种鱼类模型中央角膜厚度的比较分析。
IF 2 4区 生物学 Q1 Agricultural and Biological Sciences Pub Date : 2022-12-01 DOI: 10.1089/zeb.2022.0016
Jinpeng Li, Juan Du, Aiping Deng, Ziyang Chen, Yusong Guo, Zhongduo Wang

To understand the left-right asymmetry of vertebrate eyes, this study measured the central corneal thickness (CCT) of Oryzias curvinotus, Oryzias melastigma, Oryzias latipes, and zebrafish with optical coherence tomography. The results showed that the CCTs were significant differences among different species and groups, even between the right and left eyes of each fish. The values of the CCTs (mean ± SD, μm) for the four species were 104.71 ± 14.49, 61.88 ± 8.63, 64.76 ± 10.36 and 56.96 ± 10.48, respectively. Moreover, comparing the two wild groups of O. curvinotus from Sanya on N18° and Gaoqiao on N21°, the CCT value for the low-latitude group was 104.71 ± 14.49 μm, greater than the high latitude group 76.13 ± 5.70 μm significantly (t-test, p = 0.0001). Lastly, the paired Student's t-test revealed that significant CCT differences existed between the left and right eye for all four species and groups, of which zebrafish and O. melastigma were belonging to the left thicker type in contrast to the others. This study laid a foundation for understanding the causes of the difference in CCT, and also provided possible fish models for human researches on keratomileusis, glaucoma, and other corneal diseases.

为了了解脊椎动物眼睛的左右不对称性,本研究利用光学相干断层成像技术测量了弯曲米米(Oryzias curvinotus)、黑米米(Oryzias melastigma)、阔米米(Oryzias latipes)和斑马鱼的角膜中央厚度(CCT)。结果表明,不同种类和群体之间的cct存在显著差异,甚至在每条鱼的左右眼之间也是如此。4个物种的cct (mean±SD, μm)分别为104.71±14.49、61.88±8.63、64.76±10.36和56.96±10.48。此外,在N18°的三亚野生组和N21°的高桥野生组比较,低纬度组的CCT值为104.71±14.49 μm,显著大于高纬度组的76.13±5.70 μm (t检验,p = 0.0001)。最后,配对学生t检验显示,所有四个物种和群体的左眼和右眼之间存在显著的CCT差异,其中斑马鱼和O. melastigma属于左厚型,而其他物种属于左厚型。本研究为了解CCT差异的原因奠定了基础,也为人类角膜移植、青光眼等角膜疾病的研究提供了可能的鱼类模型。
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引用次数: 0
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