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The Effects of Plant Protein-Enriched Live Food on Larval Zebrafish Growth and the Status of Its Digestive Tract Development. 富含植物蛋白的活体食物对斑马鱼幼体生长及其消化道发育状况的影响
IF 2 4区 生物学 Q4 DEVELOPMENTAL BIOLOGY Pub Date : 2022-12-01 Epub Date: 2022-11-11 DOI: 10.1089/zeb.2022.0005
Connor W Schwepe, Michal Wojno, Giovanni S Molinari, Karolina Kwasek

Live food is necessary for the proper development of zebrafish larvae, providing nutrition in a form that is easily digestible and available to the larvae. Live food is commonly enriched to increase the dietary content of certain nutrients. However, little research has been done on protein-based enrichments, especially those of plant origin. This study sought to examine how different quality protein enrichments affected the composition of live food as well as growth and digestive tract development of larval zebrafish, Danio rerio. Larval zebrafish were fed from 3 to 22 days posthatch (dph) with one of six live food (rotifers Brachionus plicatilis and Artemia spp.) treatments: (1) live food with no enrichment (starved; control); (2) live food enriched with commercially used Spirulina spp. algae; (3) live food enriched with soybean meal (SBM); (4) live feed enriched with soy protein concentrate (SPC); (5) live feed enriched with a fishmeal hydrolysate; and (6) live feed enriched with intact fishmeal (FM). Proximate composition of live food was significantly affected by enrichment, in particular, protein content of rotifers was significantly increased by enrichment with SBM. Zebrafish fed SBM-enriched live food showed longer total body length than all other groups, except SPC. Zebrafish in the SBM group also showed increased gene expression of chymotrypsin in the intestine, possibly indicating improved intestinal development and extracellular digestion, which likely contributed to improved growth. Conversely, zebrafish fed hydrolysate-enriched live food showed reduced gene expression of alkaline phosphatase, possibly indicating a less developed intestinal tract, correlating with reduced growth compared to SBM group. Overall, plant protein was shown to be a promising source of live food enrichment for improving larval zebrafish growth.

活食是斑马鱼幼体正常发育所必需的,它以易于消化的形式为幼体提供营养。通常会对活食进行添加,以增加食物中某些营养成分的含量。然而,对以蛋白质为基础的富集物,尤其是植物来源的富集物的研究很少。本研究试图探讨不同质量的蛋白质富集物如何影响活体食物的成分以及斑马鱼幼体(Danio rerio)的生长和消化道发育。在斑马鱼幼鱼孵化后 3 到 22 天(dph)期间,用六种活体食物(轮虫 Brachionus plicatilis 和 Artemia spp.)中的一种进行喂养:(1)不添加任何添加剂的活体食物(饥饿;对照);(2)添加市售螺旋藻的活体食物;(3)添加大豆粉(SBM)的活体食物;(4)添加大豆浓缩蛋白(SPC)的活体食物;(5)添加鱼粉水解物的活体食物;(6)添加完整鱼粉(FM)的活体食物。活饵料的近似成分受到富集的显著影响,特别是轮虫的蛋白质含量在富集 SBM 后显著增加。除 SPC 组外,喂食富含 SBM 的活食的斑马鱼体长均长于其他各组。SBM 组斑马鱼肠道中糜蛋白酶的基因表达也有所增加,这可能表明肠道发育和细胞外消化得到改善,从而促进了斑马鱼的生长。相反,喂食富含水解物的活体食物的斑马鱼显示碱性磷酸酶的基因表达减少,这可能表明肠道发育较差,与 SBM 组相比,这与斑马鱼的生长减少有关。总之,植物蛋白被证明是改善斑马鱼幼鱼生长的一种很有前景的富集活食来源。
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引用次数: 0
A Simple Method for Inducing Masculinization of Zebrafish Stocks Using 17α-Methyltestosterone. 用17α-甲基睾酮诱导斑马鱼种群雄性化的简单方法
IF 2 4区 生物学 Q4 DEVELOPMENTAL BIOLOGY Pub Date : 2022-12-01 Epub Date: 2022-11-01 DOI: 10.1089/zeb.2022.0029
Jennifer L Matthews, Joy Murphy, Andrzej Nasiadka, Zoltan M Varga

Severely skewed sex ratios in zebrafish stocks can pose significant hurdles for line propagation and sperm cryopreservation. To overcome female-biased sex ratios in stocks derived from imported sperm samples, the Zebrafish International Resource Center has implemented routine supplementation of larval food with 17α-methyltestosterone to skew gonadal sex differentiation toward masculinization. Resulting stocks averaged 80% males.

斑马鱼种群的性别比例严重失衡,可能对种群繁殖和精子冷冻保存造成重大障碍。为了克服从进口精子样本中获得的种群性别比例偏雌的问题,斑马鱼国际资源中心在幼虫食物中添加了17α-甲基睾酮,以使性腺性别分化向雄性化倾斜。由此产生的种群平均80%为雄性。
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引用次数: 2
Automated Zebrafish Phenotype Pattern Recognition: 6 Years Ago, and Now. 自动斑马鱼表型模式识别:6年前和现在。
IF 2 4区 生物学 Q4 DEVELOPMENTAL BIOLOGY Pub Date : 2022-12-01 DOI: 10.1089/zeb.2022.0027
Mark Schutera, Luca Rettenberger, Markus Reischl

The article assesses the developments in automated phenotype pattern recognition: Potential spikes in classification performance, even when facing the common small-scale biomedical data set, and as a reader, you will find out about changes in the development effort and complexity for researchers and practitioners. After reading, you will be aware of the benefits and unreasonable effectiveness and ease of use of an automated end-to-end deep learning pipeline for classification tasks of biomedical perception systems.

本文评估了自动表型模式识别的发展:分类性能的潜在峰值,即使面对常见的小规模生物医学数据集,作为读者,您将发现研究人员和实践者在开发工作和复杂性方面的变化。阅读后,您将意识到用于生物医学感知系统分类任务的自动化端到端深度学习管道的好处和不合理的有效性和易用性。
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引用次数: 0
Assessment of Various Standard Fish Diets on Growth and Fecundity of Platyfish (Xiphophorus maculatus) and Medaka (Oryzias latipes). 不同标准鱼类日粮对斑螯螯虾(Xiphophorus maculatus)和斑尾螯虾(Oryzias latipes)生长和繁殖力的影响。
IF 2 4区 生物学 Q4 DEVELOPMENTAL BIOLOGY Pub Date : 2022-10-01 Epub Date: 2022-07-21 DOI: 10.1089/zeb.2022.0004
Crystal Russo, Merritt Drewery, Carolyn T Chang, Markita Savage, Lindsey Sanchez, Zoltan Varga, Michael L Kent, Ronald Walter, Yuan Lu

Several small freshwater fish species are utilized as models for human conditions and disease in biomedical research. Research animal diets are generally tailored to optimize growth, fecundity, and produce healthy research animals. However, a lack of reference diets presents a barrier in comparative studies between aquatic animal models and even among laboratories using the same species. Therefore, the objective of this study was to determine feeding regime and dietary effects on growth and fecundity in two commonly used freshwater fish, platyfish and medaka. From 1 through 6 months of age, platyfish and medaka were fed one of three feeding regime/diets: (1) our custom feeding regime consists of commercial flake food, beef liver paste, and live brine shrimp (CON); (2) a commercially available zebrafish diet, Gemma (GEM); and (3) a laboratory defined reference feeding regime (WAT). Weight, size, brood numbers, and survival rates for both species were measured monthly. Numbers of platyfish fry and hatch rate of medaka embryos were also determined. We observed that custom feeding regime (CON) fed platyfish and medaka grew larger, exhibited a higher survival rate, and had higher fecundity than WAT or GEM fed fish. These observations suggest that diets and regimes designed for zebrafish are not optimal to maintain platyfish or medaka. Thus, base diets, with clearly defined components and regimes, need to be developed with compositions that can be adjusted in a species-specific manner.

在生物医学研究中,几种小型淡水鱼类被用作人类状况和疾病的模型。研究动物的饮食通常是为了优化生长、繁殖力和生产健康的研究动物。然而,在水生动物模型之间的比较研究中,甚至在使用相同物种的实验室之间,缺乏参考饮食是一个障碍。因此,本研究的目的是确定两种常用淡水鱼,鸭嘴鱼和溪花鱼的喂养方式和饮食对其生长和繁殖力的影响。从1个月大到6个月大,鸭嘴鱼和青金石被喂食三种喂养方案/日粮中的一种:(1)我们的定制喂养方案包括商业片状食品、牛肝酱和活卤虾(CON);(2) 商业上可买到的斑马鱼饮食Gemma(GEM);和(3)实验室定义的参考饲养制度(WAT)。两个物种的体重、大小、繁殖数量和存活率每月进行测量。还测定了鸭嘴鱼鱼苗的数量和花呢鱼胚胎的孵化率。我们观察到,与WAT或GEM喂养的鱼类相比,定制喂养制度(CON)喂养的鸭嘴鱼和花呢鱼体型更大,存活率更高,繁殖力更高。这些观察结果表明,为斑马鱼设计的饮食和制度并不是维持鸭嘴鱼或墨鱼的最佳选择。因此,需要制定成分和制度明确的基础饮食,其成分可以以特定物种的方式进行调整。
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引用次数: 1
Evaluation of Media Conductivity and a Combination of Iodine and Sodium Hypochlorite Surface Disinfection on Zebrafish (Danio rerio) Embryo Viability and Morphology. 介质电导率和碘和次氯酸钠联合表面消毒对斑马鱼胚胎活力和形态的评估。
IF 2 4区 生物学 Q4 DEVELOPMENTAL BIOLOGY Pub Date : 2022-10-01 Epub Date: 2022-10-07 DOI: 10.1089/zeb.2022.0028
Gerardo Mendoza, Adedeji A Afolalu, Kvin Lertpiriyapong, Neil S Lipman, Christine Lieggi

Embryo surface disinfection in either an iodine or sodium hypochlorite (NaOCl) solution is commonly performed on imported zebrafish embryos to decrease pathogen introduction into a facility. The impact of the consecutive use of these disinfectants and the conductivity of the culture media on embryo survival and development post-disinfection have not been evaluated. Iodine (12.5-25 ppm) is effective at eliminating several Mycobacterium species, whereas NaOCl (50-100 ppm) reduces the number of viable Pseudoloma neurophilia spores. Casper and T5D (tropical 5D wild type) embryos reared in media of differing conductivities (0-10, 100-200, 750-950, and 1500-2000 μS) with and without exposure to NaOCl 100 ppm at 6 h post-fertilization were evaluated for survival, hatching success, and morphological defects at 5 days post-fertilization. Additionally, the consecutive use of iodine (12.5 ppm for 2 min) followed by NaOCl (75 or 100 ppm for 10 min), as well as the inverse, was evaluated. Embryo survival was not impacted by embryo rearing media alone; however, survival significantly decreased when embryos were disinfected with 100 ppm NaOCl in media with a conductivity >750-950 μS. Iodine (12 ppm) and NaOCl (75 ppm) used sequentially resulted in >50% survival, whereas the use of 100 ppm NaOCl resulted in high levels of embryo mortality.

通常在碘或次氯酸钠(NaOCl)溶液中对进口斑马鱼胚胎进行胚胎表面消毒,以减少病原体引入设施。连续使用这些消毒剂和培养基的导电性对消毒后胚胎存活和发育的影响尚未评估。碘(12.5-25 ppm)能有效地消灭几种分枝杆菌,而NaOCl(50-100 ppm)减少了有活力的亲神经假瘤孢子的数量。Casper和T5D(热带5D野生型)胚胎在不同电导率(0-10、100-200、750-950和1500-2000)的培养基中饲养 μS)暴露于和不暴露于NaOCl 100 6时的ppm 对受精后h的存活率、孵化成功率和受精后5天的形态缺陷进行评估。此外,连续使用碘(12.5 2的ppm 分钟),然后加入NaOCl(75或100 10的ppm min),以及倒数。胚胎存活不受单独培养基的影响;然而,当用100消毒胚胎时,存活率显著下降 电导率>750-950的介质中的NaOCl ppm μS。碘(12 ppm)和NaOCl(75 ppm)导致>50%的存活率,而使用100 ppm的NaOCl导致高水平的胚胎死亡率。
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引用次数: 0
The Need for a Framework Addressing the Temporal Aspects of Fish Sperm Motility Leading to Community-Level Standardization. 需要一个框架解决鱼类精子运动的时间方面导致社区水平的标准化。
IF 2 4区 生物学 Q4 DEVELOPMENTAL BIOLOGY Pub Date : 2022-08-01 DOI: 10.1089/zeb.2022.0006
Harvey Blackburn, Leticia Torres, Yue Liu, Terrence R Tiersch

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 (Danio rerio) 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.

活力是一个广泛可用的参数,可用于评估水生物种的精子质量。体外受精鱼类的精子在激活后通常经历一个动态的、短暂的运动期。在任意运动峰值处分配单一值的常见做法给可重复性、社区水平标准化和跨研究比较带来了挑战。本研究旨在探索标准化运动报告的统计方法,并为社区标准制定初步框架。采用计算机辅助精子分析方法,对14条斑马鱼(Danio rerio)共21,705个细胞的精子样本进行分析,在激活后10 s开始收集数据,每5 s间隔50 s。选择四个常见的运动变量进行分析:曲线速度,直线速度,节拍交叉频率和横向头部位移幅度。聚类分析用于评估男性内部和男性之间的精子亚群随时间的变化,最小二乘法用于探索时间方面,回归方程的一阶导数用于计算运动参数的变化率。聚类分析证明信息丰富,但重叠的短暂聚类对于提供标准化选项没有价值。对时间方面和变化率的分析表明,通过报告总体运动性-时间函数或在稳定的时间窗口而不是峰值运动性或随机时间报告,可以实现标准化。这些方法可以在提供可比信息的同时,最大限度地减少雄性间变异和亚种群动态变化引起的不一致性。确定了收集-处理-冷冻保存-解冻序列的动态报告的总体时间框架,为支持社区层面的标准化工作提供了基础。
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引用次数: 4
Simple and High-Throughput Rheotaxis Behavioral Assay for Zebrafish Larva. 斑马鱼幼体简单、高通量致流变行为测定。
IF 2 4区 生物学 Q4 DEVELOPMENTAL BIOLOGY Pub Date : 2022-06-03 DOI: 10.1089/zeb.2021.0087
M. Tantry, V. S. Harini, Kirankumar Santhakumar
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.
斑马鱼(Danio rerio)被用作研究感音神经性听力损失的模型。对毛细胞的损伤可以通过对斑马鱼的流变行为进行评分来评估。在这项研究中,我们新设计了一种能够量化斑马鱼幼虫流变性行为的流变性行为测定方案。我们使用硫酸铜(一种众所周知的耳毒素)对幼虫进行化学诱导耳毒性,并测定了不同流速下的流变性。简单、经济高效、高通量的流变性测定系统可以为药物开发和其他行为研究提供深入的见解。
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引用次数: 1
Identification of Transcription Factor Binding Sites by Cleavage Under Target and Release Using Nuclease in Zebrafish 斑马鱼靶下裂解和核酸酶释放鉴定转录因子结合位点
IF 2 4区 生物学 Q4 DEVELOPMENTAL BIOLOGY Pub Date : 2022-06-01 DOI: 10.1089/zeb.2021.0082
Radwa A. Barakat, C. Campbell, Raquel Espín-Palazón
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.
利用核酸酶在靶下切割和释放(CUT&RUN)已经成为一种超越传统方法的染色质分析策略。尽管CUT&RUN已经在哺乳动物细胞中得到了广泛的应用,但它在斑马鱼中的应用还处于早期阶段。在这项研究中,我们开发了一种方案,成功地执行CUT&RUN来绘制胚胎、成年组织和facs分类的斑马鱼细胞中的转录因子(TF)结合位点。我们还提供了一个详细的工作流程,用于识别可用于任何动物物种的预测TF结合位点。总之,我们的策略将把这个宝贵的工具扩展到斑马鱼群落,提高这种模式生物的表观遗传分辨率。
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引用次数: 2
Dopamine Level Affects Social Interaction and Color Preference Possibly Through Intestinal Microbiota in Zebrafish. 多巴胺水平可能通过斑马鱼肠道微生物群影响社会互动和颜色偏好。
IF 2 4区 生物学 Q4 DEVELOPMENTAL BIOLOGY Pub Date : 2022-06-01 DOI: 10.1089/zeb.2021.0077
Ju Wang, Feng Zheng, Lifen Yin, Shengnan Shi, Bing Hu, Hao Qu, Lei Zheng
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.
越来越多的研究表明,胃肠系统中的微生物群会影响大脑的神经发育和行为程序。然而,强调浅滩行为与肠道微生物群之间关系的机制仍然存在争议,负责任的神经递质的作用仍然不清楚。在这里,我们发现浅滩行为影响了浅滩的颜色偏好,表明浅滩倾向于选择有利于社会接触的有利颜色环境。同时,选择性D1-R拮抗剂SCH23390的给药可能会破坏导致浅滩颜色偏好缺陷的社会互动。更重要的是,抗生素土霉素(OTC)暴露导致的微生物群改变降低了社交能力,削弱了浅滩对所有颜色组合的偏好。当在OTC暴露后补充鼠李糖乳杆菌GG时,鱼类保持了与正常鱼类相同的社会凝聚力和颜色偏好能力。我们的研究结果支持多巴胺在塑造浅滩颜色偏好中的作用。我们的研究结果表明,大脑的多巴胺水平可以介导社会识别和颜色偏好,并提供了多巴胺的产生通过肠道微生物群协调的可能性。
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引用次数: 1
A New Transgenic Line for Rapid and Complete Neutrophil Ablation. 一种快速完全消融中性粒细胞的转基因新品系。
IF 2 4区 生物学 Q4 DEVELOPMENTAL BIOLOGY Pub Date : 2022-06-01 DOI: 10.1089/zeb.2022.0020
Christopher J Hall, Jonathan W Astin, Jeff S Mumm, David F Ackerley

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.

斑马鱼细胞系在特定的细胞区室中表达硝基还原酶(NTR),使这些细胞对甲硝唑(MTZ)介导的消融敏感,已被证明在研究组织再生和研究细胞功能方面非常有用。与许多细胞相比,中性粒细胞对NTR/MTZ靶向消融策略具有相对抗性。最近,一种合理设计的NTR变体(NTR 2.0)在斑马鱼中显示出极大改善的mtz介导的消融效果。我们发现,中性粒细胞限制性表达NTR 2.0的转基因细胞系显示中性粒细胞完全消融,MTZ剂量比当前治疗方案少100倍,治疗时间短至5小时。
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引用次数: 3
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Zebrafish
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