Zebrafish最新文献

This study aimed to evaluate the viability of gametes in zebrafish (Danio rerio), at different rigor mortis stages. Viability assessments were conducted on oocytes at various developmental stages using LIVE/DEAD and the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction assay. For sperm evaluation, both kinetic (computer-assisted sperm analysis) and morphological assessments (Rose Bengal staining) were performed. Results demonstrated that rigor mortis progression significantly impacted oocyte viability during post-rigor stages, with the following viability rates: pre-rigor (70.43 ± 12.31%), fresh/control (46.43 ± 12.54%), post-rigor (27.62 ± 22.29%), and rigor mortis (comparable to fresh/control). Conversely, sperm kinetics exhibited nuanced responses to the rigor mortis stages, with specific parameters showing sensitivity, whereas the others remained relatively stable. Sperm motility was higher in the fresh/control (63.23 ± 19.03%) and pre-rigor (58.96 ± 14.38%) compared to the post-rigor group (3.34 ± 4.65%). This study highlights the significance of the pre-rigor for successful gamete collection and preservation. These findings provide valuable insights for conservation efforts and optimization of genetic resource management for endangered fish species. This study aimed to develop effective assistive reproductive techniques by elucidating the interplay between rigor mortis and gamete quality, contributing to the broader goals of species conservation and maintenance of genetic diversity in fish populations.

The small characins represent a systematic puzzle in the Neotropical ichthyofauna as a result of independent miniaturization processes, adaptive convergence and lack of diagnostic characters for several genera. In order to diminish the taxonomic uncertainties and the evolutionary pathways in Hemigrammus, we carried out an integrative genetic analysis in the putatively widespread Hemigrammus marginatus Ellis, 1958 by combining cytogenetic and molecular data based on the mitochondrial Cytochrome C Oxidase subunit I (COI). Specimens of H. marginatus from the type locality in Itapicuru River basin and other two populations from coastal rivers in northeastern Brazil were analyzed and compared with the available data from other regions in South America. Conspicuous macro and microkaryotypic differences were detected between the samples from northeastern and southern Brazil (Upper Paraná River basin). Likewise, the DNA barcoding and species delimitation analyses recovered distinct Molecular Operational Taxonomical Units within H. marginatus. Therefore, the population from the type locality should be referred to as H. marginatus stricto sensu, representing a restricted characin taxon from coastal drainages (including the São Francisco River basin) along northeastern Brazil, while other populations of this small characin fish need to be taxonomically revised and managed as unique lineages.

Despite decades of research with laboratory rodents, the mechanisms underlying learning and memory, and their impairment, are still not fully understood. The zebrafish is a newcomer in this research area, but it has shown great promise. Food is often employed as a reinforcer in learning tasks with rodents. However, for zebrafish, food has been a problematic reinforcer. Controlling timing and localization of its delivery is difficult. What food types zebrafish prefer is also rarely studied? Here, we describe a novel food delivery hardware and procedure. The apparatus is simple, cheap to manufacture, and easy to employ. Using this new method, we compare how zebrafish respond to three food types, artemia nauplii, crushed tropical fish flakes, and small zebrafish pellets. In binary choice tasks, we show that zebrafish spend significantly more time near the artemia delivery cylinder, swim closer to, and visit this cylinder more frequently compared to food cylinders delivering flakes or pellets, while responses to these latter two cylinders do not differ from each other. We conclude that the newly developed method allows the quantification of food preference in zebrafish, and that it will lead to the identification of highly rewarding food types for learning studies in this species.

Zebrafish larvae are used to model the pathogenesis of multiple bacteria. This transparent model offers the unique advantage of allowing quantification of fluorescent bacterial burdens (fluorescent pixel counts [FPC]) in vivo by facile microscopical methods, replacing enumeration of bacteria using time-intensive plating of lysates on bacteriological media. Accurate FPC measurements require laborious manual image processing to mark the outside borders of the animals so as to delineate the bacteria inside the animals from those in the culture medium that they are in. Here, we have developed an automated ImageJ/Fiji-based macro that accurately detects the outside borders of Mycobacterium marinum-infected larvae.

Photoperiod and temperature are two of the most powerful environmental cues that entrain circadian clocks. Being ectothermic, fish must keep their body temperature within a physiological range to optimize biological processes mainly applying behavioral strategies. Here, we developed a low-cost, automated system that allows to create a horizontal multiple-step thermal gradient and video record fish behavior for long-term periods. To validate the system, we assessed daily thermal preference and locomotor activity in the teleost Danio rerio. Our apparatus provides the opportunity to investigate the behavioral thermoregulation of captive fish, and our results highlight the importance of considering thermal preferences when designing husbandry protocols.

Intestinal permeability plays a crucial role in intestinal barrier function. Altered intestinal permeability is well documented in numerous chronic diseases and may serve as a risk factor for disease onset as well as a target for innovative therapeutic strategies. While reliable and sensitive approaches for studying intestinal permeability have been established in animal models, such as mice and zebrafish larvae, methods for investigating this in adult zebrafish remain a considerable challenge. The zebrafish has emerged as a valuable model for studying intestinal development, physiology, and disease. Moreover, zebrafish offer certain advantages over rodent models, such as the ability to evaluate the dynamic interactions of labeled markers in vivo and in real time. In this study, we present a comprehensive pipeline for assessing in vivo intestinal permeability in adult zebrafish using fluorescent-labeled dextran. Detailed protocols for fish handling, reagent preparation, optimization of reagent dosage and delivery routes, and quantification of fluorescent markers in extraintestinal sites are provided. Our findings suggest that zebrafish hold promise as an alternative model for in vivo investigations of intestinal permeability induced by genetic, pathophysiological, and/or pharmacological events.

Retinal markers with high quality and specificity are important for the observation of pathologic changes of retinal cells during retinal development, degeneration, and regeneration. The zpr-3 antibody is widely used to label rods in zebrafish, but the exact antigen is still unknown. In this study, we provided evidence to demonstrate that the antigen gene of zpr-3 is rho, which encodes the rod opsin, and the exact epitope of zpr-3 is the 320-354 region of Rho protein. More importantly, our immunofluorescence assays indicated that zpr-3 labels both the outer segments of rods and green cones on zebrafish retinal sections, probably due to the cross-reaction with the green-cone opsin. Our work is valuable for the scientific community to interpret the experimental data involving the zpr-3 antibody.

Hydrocephalus is a cerebrospinal fluid-related disease that usually manifests as abnormal dilation of the ventricles, with a triad of clinical findings including walking difficulty, reduced attention span, and urinary frequency or incontinence. The onset of congenital hydrocephalus is closely related to mutations in genes that regulate brain development. Currently, our understanding of the mechanisms of congenital hydrocephalus remains limited, and the prognosis of existing treatments is unsatisfactory. Additionally, there are no suitable or dedicated model organisms for congenital hydrocephalus. Therefore, it is significant to determine the mechanism and develop special animal models of congenital hydrocephalus. Recently, zebrafish have emerged as a popular model organism in many fields, including developmental biology, genetics, and toxicology. Its genome shares high similarity with that of humans, and it has fast and low-cost reproduction. These advantages make it suitable for studying the pathogenesis and therapeutic approaches for various diseases, specifically congenital diseases. This study explored the possibility of using zebrafish as a model organism for congenital hydrocephalus. This review describes the characteristics of zebrafish and discusses specific congenital hydrocephalus models. The advantages and limitations of using zebrafish for hydrocephalus research are highlighted, and insights for further model development are provided.

The 2024 Zebrafish Husbandry Workshop and Summit held during the World Aquaculture Society Meeting in San Antonio focused on key areas for improving zebrafish husbandry research. Discussions highlighted the need for comprehensive literature on husbandry, better communication and collaboration between researchers and facility staff, and the adoption of a standardized reference diet. Current literature lacks comprehensive data and often overlooks crucial factors such as housing density and space requirements for fish development. Collaborative efforts between researchers and facility managers are essential for acquiring accurate husbandry data and minimizing pathogen risks. Standardizing descriptive language and parameter lists in publications and enhancing communication between facilities can improve research quality. Action items proposed include better communication of incoming fish information, standardization of pathogen monitors, transparency in husbandry practices, and fostering a spirit of collaboration among organizations. The summit emphasized the importance of increased PI awareness about husbandry, testing existing standardized diets, forming consortia to oversee diet standardization, creating unified repositories and forums, and conducting evidence-based husbandry studies.