Zebrafish最新文献

Low and variable growth and survival rates (SR) of 6-10 days postfertilization zebrafish larvae are a problem. This problem seems to be linked to starter feed characteristics. This study is an attempt to find alternatives to address these requests. For this, larvae were fed fresh and lyophilized microalgae (Chlorella, Scenedesmus, and Haematococcus), egg yolk (YOLK), lyophilized Artemia nauplii (LAN), and a combination of them. The lowest SR was observed in algae-fed larvae. All died on day 11 showing an emaciated appearance, similar to starved larvae. The highest SR was observed in YOLK- and LAN-fed larvae, which also showed an elongated anterior part of the body. Negative correlations of SR with vegfaa (vascular endothelial growth factor) and morphometric traits with igf2a (insulin-like growth factor) were also found and supported by changes at the molecular level. The presence of algae in the digestive tract of the larvae and the observation of fecal droppings indicate that the algae have an appropriate size and are palatable. The increase in the digestive enzyme activity shows the larval effort to digest the algae. The fact that the algae-fed larvae died even before the larvae were kept in starvation indicates the dramatic amount of energy that the larvae spent in microalgae digestion. Although both YOLK- and LAN-fed larvae had the highest SR, LAN group started to feed on Artemia nauplii sooner. This can be linked to the delayed growth in YOLK-fed larvae and an accelerated growth in the case of LAN-fed group. LAN is an expensive feed with negative effects on water quality, whereas YOLK is a cheap and nutritionally balanced feed with fine granular texture that contributes to a larval SR similar to LAN without affecting water quality. In conclusion, microalgae cannot be considered a suitable starter food for zebrafish, whereas LAN and YOLK can be considered good starter feeds.

After its introduction for scientific investigation in the 1950s, the cypriniform zebrafish, Danio rerio, has become a valuable model for the study of regenerative processes and mechanisms. Zebrafish exhibit epimorphic regeneration, in which a nondifferentiated cell mass formed after amputation is able to fully regenerate lost tissue such as limbs, heart muscle, brain, retina, and spinal cord. The process of limb regeneration in zebrafish comprises several stages characterized by the activation of specific signaling pathways and gene expression. We review current research on key factors in limb regeneration using zebrafish as a model.

The zebrafish (Danio rerio) is an emerging model system in several research areas worldwide, especially in the Global South. In this context, the present study revised the historical use and trends of zebrafish as experimental models in Brazil. The data concerning the bibliometric parameters, research areas, geographic distribution, experimental design, zebrafish strain, and reporter lines, as well as recent advances were revised. In addition, the comparative trends of Brazilian and global research were discussed. Revised data showed the rapid growth of Brazilian scientific production using zebrafish as a model, especially in three main research areas (Neuroscience &and Behavior, Pharmacology and Toxicology, and Environment/Ecology). Studies were conducted in 19 Brazilian states (70.37%), confirming the wide geographic distribution and importance of zebrafish research. Results indicated that research related to toxicological approaches are widespread in Global South countries such as Brazil. Studies were performed mainly using in vivo tests (89.58%) with adult fish (59.75%) and embryos (30.67%). Moreover, significant research gaps and recommendations for future research are presented. The present study shows that the zebrafish is a suitable vertebrate model system in the Global South.

This study was aimed to assess effects of three strains of probiotics Lactobacillus acidophilus NCFM, Lactobacillus rhamnosus HN001, and Bifidobacterium animalis subsp. lactis Bi-07 on the intestinal motility and inflammation in the zebrafish models. The intestinal motility model was established using 5 days postfertilization (dpf) zebrafish administered with a fluorescent dye Nile red at 10 ng/mL for 16 h, followed by probiotics treatment for 24 h and the intestinal motility was inversely proportional to the intestinal fluorescence intensity that was quantitatively measured by image analysis. The intestinal inflammation was induced by treating 3 dpf neutrophil fluorescent zebrafish with 0.0125% of trinitrobenzenesulfonic acid for 48 h. Probiotics were administered at low, moderate, and high concentrations determined based on maximum tolerable concentration through soaking. All three strains of probiotics promoted intestinal movement, of which B. animalis subsp. lactis Bi-07 was most potent at lower concentrations. L. rhamnosus HN001 and B. animalis subsp. lactis Bi-07 had the therapeutic effects on the intestinal inflammation and the inflammation-associated mucosal damage recovery. The anti-inflammatory mechanisms of L. rhamnosus HN001 was related to both reduce inflammatory factor interleukin-6 (IL-6) and restored tissue repair factor transforming growth factor-β-1 (TGFβ-1); whereas B. animalis subsp. lactis Bi-07 was probably only associated with TGFβ-1 elevation. Using larval zebrafish models for probiotics screening and assessment would speed up product research and development and improve products' efficacy and quality.

The melanocortin system is a key structure in the regulation of energy balance. Overexpression of inverse agonists, agouti-signaling protein (ASIP), and agouti-related protein (AGRP) results in increased food intake, linear growth, and body weight. ASIP regulates dorsal-ventral pigment polarity through melanocortin 1 receptor (MC1R) and overexpression induces obesity in mice by binding to central MC4R. Asip1 overexpression in transgenic zebrafish (asip1-Tg) enhances growth, yet experiments show fish overexpressing Asip1 do not develop obesity even under severe feeding regimes. Asip1-Tg fish do not need to eat more to grow larger and faster; thus, increased food efficiency can be observed. In addition, asip1-Tg fish reared at high density are able to grow far more than wild-type (WT) fish reared at low density, although asip1-Tg fish seem to be more sensitive to crowding stress than WT fish, thus making the melanocortin system a target for sustainable aquaculture, especially as the U.S. Food and Drug Association has recently approved transgenic fish trading.

The traditional thesis of the osmorespiratory compromise is that low branchial water and ion permeability would be traded off for increased O₂ permeability at times of elevated O₂ demand. However, there is growing evidence of independent regulation of these permeabilities in hypoxia-tolerant fish. Using 0.5-g zebrafish previously maintained under normoxia at 25°C, we investigated responses to acute temperature challenges (15°C or 35°C), acute hypoxia (15 min at 10% or 5% air saturation), as well as longer-term exposures to 10% hypoxia, on O₂ consumption (MO₂), diffusive water flux, and net sodium loss rates. Exposure to 35°C increased, and 15°C decreased all three rates, with diffusive water flux showing the lowest temperature sensitivity, and Na⁺ loss the greatest. Acute 10% and 5% hypoxia increased diffusive water flux and net Na⁺ loss, and it reduced MO₂. All these responses reflected the traditional osmorespiratory compromise. However, during prolonged 10% hypoxia, MO₂ recovered, diffusive water flux decreased below control levels, and Na⁺ loss rate remained elevated, even during posthypoxia recovery. Overall, zebrafish do not fit standard patterns previously seen in either hypoxia-tolerant or -intolerant fish but are clearly able to adjust the effective permeabilities of their gills to O₂, water, and ions independently during acute temperature and hypoxia exposures.

A modified protocol for the extraction and analysis of cortisol in individual zebrafish, Danio rerio, embryo, and larva samples has been developed and evaluated. Recovery efficiency of the method was high, specifically calculated at 93.8% ± 6.5%. Dilution tests showed high parallelism, while increasing the number of individuals used in each extraction sample resulted in a linear, although slightly underestimated, increase of cortisol yield. Results of cortisol content from 0, 3, and 5 days postfertilization (dpf) fish using the proposed protocol were within the range of most published studies analyzing cortisol in pooled samples of 10-30 individuals. Moreover, 5 dpf larvae had significantly higher cortisol levels than embryos, a pattern commonly observed in literature. Finally, application of an osmotic stress in 5 dpf larvae led to a statistically significant increase in cortisol content.

Rapidly accumulating literature has proven feasibility of the zebrafish xenograft models in cancer research. Nevertheless, online databases for searching the current zebrafish xenograft literature are in great demand. Herein, we have developed a manually curated database, called ZenoFishDb v1.1 (https://konulab.shinyapps.io/zenofishdb), based on R Shiny platform aiming to provide searchable information on ever increasing collection of zebrafish studies for cancer cell line transplantation and patient-derived xenografts (PDXs). ZenoFishDb v1.1 user interface contains four modules: DataTable, Visualization, PDX Details, and PDX Charts. The DataTable and Visualization pages represent xenograft study details, including injected cell lines, PDX injections, molecular modifications of cell lines, zebrafish strains, as well as technical aspects of the xenotransplantation procedures in table, bar, and/or pie chart formats. The PDX Details module provides comprehensive information on the patient details in table format and can be searched and visualized. Overall, ZenoFishDb v1.1 enables researchers to effectively search, list, and visualize different technical and biological attributes of zebrafish xenotransplantation studies particularly focusing on the new trends that make use of reporters, RNA interference, overexpression, or mutant gene constructs of transplanted cancer cells, stem cells, and PDXs, as well as distinguished host modifications.

Collecting large numbers of rare cells for high-throughput molecular analysis remains a technical challenge, primarily due to limitations in existing technologies. In developmental biology this has impeded single-cell analysis of primordial organs, which derive from few cells. In this study, we share novel transgenic lines for rapid cell enrichment from zebrafish embryos using human surface antigens for immunological binding and magnetic sorting. As proof of principle, we tagged, enriched, and performed single-cell RNA sequencing on nascent hematopoietic stem/progenitor cells and endothelial cells from early embryos. Our method is a quick, efficient, and cost-effective approach to a previously intractable problem.

Black Soldier Fly (BSF) meal is considered an alternative, emerging, and sustainable ingredient for aquafeed formulation. However, results on fish physiological responses are still fragmentary and often controversial, and no data are available on the effect of insect meal-based diets on fish reproduction. On this regard, zebrafish, with its relatively short life cycle, represents an ideal experimental model to explore this topic. In this study, female zebrafish were fed for 12 months on a control diet based on fish meal (FM) and fish oil and two experimental diets with full-fat BSF (Hermetia illucens) prepupae meal inclusion, to replace 25% and 50% of FM (BSF25 and BSF50). All diets were isonitrogenous, isolipidic, and isoenergetic. The effects of these two experimental diets on female's reproduction were investigated through a multidisciplinary approach, including the evaluation of growth, gonadosomatic index, spawned/fertilized eggs and hatching rate, adult female carcass and fertilized egg fatty acid composition, histological analysis of the ovary, spectroscopic macromolecular composition of class IV oocytes, and expression of genes involved in fish lipid metabolism in the liver. Results showed that while fish were perfectly able to cope with a 25% insect meal dietary inclusion, a 50% inclusion level caused the overexpression of genes involved in lipid metabolism, a general reduction in the number of spawned eggs, and differences in the frequency rate of previtellogenic oocytes, class III, IV, oocytes and postovulatory follicles and atretic oocytes, in the macromolecular composition of class IV oocytes, and in the fatty acid composition of the fertilized eggs, respect to control and 25% group.