Zebrafish最新文献

Animal models are an important tool for studying noncommunicable diseases (NCDs) as they provide a unique opportunity to investigate real-time changes that occur in the onset of, and during, the diseased state. This is of particular importance given that the global prevalence of NCDs, such as type 2 diabetes mellitus (T2DM), is rising at an alarming rate. In South Africa, which has one of the highest levels of HIV in the world, the incidence of T2DM is thought to be associated, in part, with exposure to combination antiretrovirals. We report on the establishment of both nonobese and obese zebrafish models of T2DM, as well as associated changes in mRNA expression of preproinsulin and phosphoenolpyruvate carboxykinase (pck) 1 and 2. The diabetic state was achieved by either immersing adult zebrafish in a 2% glucose solution for 40 days or by overfeeding adult zebrafish for 10 weeks. Glucose immersion resulted in significantly elevated fasting blood glucose levels twice as high as control, whereas bodyweight did not change significantly (nonobese model). Overfeeding led to both significantly elevated fasting blood glucose and bodyweight compared with control (obese model). Both models were characterized by significantly increased preproinsulin mRNA expression indicating insulin resistance; mRNA expression of metabolic enzymes PCK 1 and 2 was also significantly upregulated, as seen in diabetic patients. These candidate gene expression changes, similar in both zebrafish models, establish a baseline that can be utilized to investigate the underlying mechanisms driving the increased T2DM incidence, using an excellent alternative to traditional rodent models.

Increasing carbon dioxide levels associated with climate change will likely have a devastating effect on aquatic ecosystems. Aquatic environments sequester carbon dioxide, resulting in acidic conditions that can negatively affect fish development. Increasing climate change impacts in the coming decades will have an outsized effect on younger generations. Therefore, our research had two interconnected goals: 1) understand how aquatic acidification affects the development of zebrafish, and 2) support a high school scientist's ability to address environmental questions of increasing importance to her generation. Working with teachers and other mentors, the first author designed and conducted the research, first in her high school, then in a university research laboratory. Zebrafish embryos were reared in varying pH conditions (6.7-8.2) for up to 7 days. We assessed fish length and development of the inner ear, including the otoliths; structures that depend on calcium carbonate for proper development. Although pH did not affect fish length, fish reared in pH 7.75 had smaller anterior otoliths, showing that pH can impact zebrafish ear development. Furthermore, we demonstrate how zebrafish may be used for high school students to pursue open-ended questions using different levels of available resources.

Xenotransplantation of neuroblastoma cells into larval zebrafish allows the characterization of their in vivo tumorigenic abilities and high-throughput treatment screening. This established preclinical model traditionally relies on microinjection into the yolk or perivitelline space, leaving the engraftment ability of cells at the hindbrain ventricle (HBV) and pericardial space (PCS), sites valuable for evaluating metastasis, angiogenesis, and the brain microenvironment, unknown. To address this gap in knowledge, Casper zebrafish at 48 h postfertilization were microinjected with approximately 200 Kelly, Be(2)-C, SK-N-AS, or SY5Y cells into either the HBV or PCS. Fish were imaged at 1, 3, and 6 days postinjection and tumor growth was monitored at each timepoint. We hypothesized that engraftment ability and location preference would be cell line dependent. Kelly and SK-N-AS cells were able to engraft at both the HBV and PCS, with a near doubling in size of tumor volume during the 6 days observation period, with cells appearing to grow better in the HBV. Be(2)-C tumors remained static while SY5Y tumors decreased in size, with almost complete loss of volume at both sites. Therefore, the capability of neuroblastoma cell engraftment in zebrafish larvae is cell line dependent with a location preference.

Soybean meal (SBM) has become a common dietary replacement for fish meal (FM) in aquafeed. However, at high inclusions, SBM has been shown to have negative impacts presenting as reduced feed intake and intestinal inflammation. Medicinal plant extracts, namely essential oils, have been used to promote growth performance and immune response. The objective of this study was to investigate the potential therapeutic effects of oregano (Origanum vulgare) essential oil (OEO) inclusion on utilization of a high-inclusion SBM diet using zebrafish as a model. Five diets were used in this study: reference-FM-based diet, control-55.7% inclusion SBM diet, and three experimental SBM-based diets OEO1, OEO2, and OEO3 that were supplemented with 1%, 2%, or 3% of oregano oil, respectively. The FM group had overall better growth performance when compared with the other treatment groups; however, the OEO3 mean weight and feed conversion ratio were not significantly different from the FM group (p > 0.05) and were significantly improved compared with the SBM group (p < 0.05). Similarly, OEO2 total length was not significantly different from FM (p > 0.05) but significantly higher than the SBM group (p < 0.05). Expression of inflammation-related genes did not significantly differ between the OEO groups and the SBM-only group. However, the OEO2 and OEO3 groups displayed improved growth performance compared with the SBM group, suggesting that inclusion of OEO at or above 2% inclusion may help to alleviate common symptoms induced by a high-inclusion SBM diet.

Various methods have been used in rodents to evaluate learning and memory. Although much less frequently used, the zebrafish emerges as an alternative model organism in this context. For example, it allows assessing potential behavioral deficits because of neurodevelopmental disorders or environmental neurotoxins. A variety of learning tasks have been employed in previous studies that required extensive habituation and training sessions. Here, we introduce a simpler and faster method to evaluate learning and memory of zebrafish with minimum habituation. A new apparatus, a transparent L-shaped tube, was developed in which we trained each zebrafish to swim through a long arm and measured the time to swim through this arm. We demonstrate that in this task, zebrafish could acquire both short-term (1 h) and long-term memory (4 days). We also studied learning and memory of a gene knockout (KO) zebrafish that showed social impairments related to autism. We found KO mutant zebrafish to show a quantitative impairment in habituation, learning, and memory performance compared with wild-type control fish. In conclusion, we established a novel learning apparatus and sensitive paradigm that allowed us to evaluate learning and memory of adult zebrafish that required only a brief habituation period and minimal training.

Over the years, scientific research with fish models has grown at a rapid pace, and issues such as animal welfare are becoming increasingly important in various areas of animal husbandry and experimentation. Here, we evaluated whether Danio rerio behavior is affected by long-term maintenance (75 days) in an enriched environment or a chronic stress (CS) situation. In addition, we evaluated some biochemical parameters related to redox status. We concluded that long-term maintenance of zebrafish in enriched environment might induce an anxiety-like behavior pattern when these fish are faced with an acute subsequent stressor. These anxiety results, the increased school cohesion, and the absence of oxidative damage allow us to hypothesize that the fish maintained in environmental enrichment (EE) situation is more reactive, showing a strong protective reaction to the stress. From an applicable perspective, we show that both too much stress and too little stress are not ideal for zebrafish stocks. In CS situations, fish can habituate and might not respond optimally to test conditions. In opposite, the low stress promoted by environmental enrichment also renders the fish incapable of dealing with occasional stressors optimally, because now even normal conditions appear stressful to them and may elicit fear behaviors they normally would not exhibit.

Using integrative tools can be effective for species identification, especially in complex groups like Astyanax. Astyanax bimaculatus group is composed of six valid species, including A. lacustris. "A. altiparanae", "A. asuncionensis", and "A. jacuhiensis" are considered as junior synonyms of A. lacustris. Seeking to test the operational taxonomic unit (OTU) status of the junior synonyms of A. lacustris ("A. altiparanae", "A. asuncionensis", and "A. jacuhiensis"), we used analyses through mitochondrial DNA (COI and Cytb), cytogenetic markers (classical and molecular), and morphometry ("truss network"). Analysis of mitochondrial DNA sequences separated A. lacustris from the other synonymized species. The cytogenetic and morphometric analyses did not corroborate the synonymization and suggest that besides A. lacustris, the OTUs A. altiparanae, A. asuncionensis, and A. jacuhiensis are valid species. The analysis of different characters proposed by the integrative taxonomy used on the same individuals could provide greater reliability and minimize the underestimation of biodiversity.

Global warming and extreme weather events pose a significant threat to global biodiversity, with rising water temperatures exerting a profound influence on fish conservation and fishery development. In this study, we used zebrafish as a model organism to explore the impact of a heat acclimation period on their survival rates. The results demonstrated that a 2-month heat acclimation period almost completely mitigated heat stress-induced mortality in zebrafish. Subsequent analysis of the surviving zebrafish revealed a predominance of hepatic mitochondria in a fission state. Remarkably, a short-term fasting regimen, which induced hepatic mitochondrial fission, mirrored the outcomes of the protective effect of heat acclimation and augmented animal survival under heat stress. Conversely, treatment with a mitochondrial fission inhibitor within the fasting group attenuated the elevated survival rate. Furthermore, zebrafish embryos subjected to brief heat acclimation also exhibited increased heat resistance, a trait diminished by a chemical intervention inhibiting mitochondrial fission. This suggests a shared mechanism for heat resistance between embryos and adult zebrafish. These findings underscore the potential use of inducing mitochondrial fission to enhance heat resistance in zebrafish, offering promise for fish biodiversity conservation in the face of global warming.

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 nucleoplasmin 2b (npm2b), a female-specific essential maternal effect gene, causing heritable sex-limited disruption of reproduction. HMEL-induced high-efficiency mutation of npm2b in females suppresses population, while males transmit the HMEL 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.