scholarly journals Standardized Method for the Assessment of Behavioral Responses of Zebrafish Larvae

Biomedicines ◽  
2021 ◽  
Vol 9 (8) ◽  
pp. 884
Author(s):  
Hideyuki Maeda ◽  
Noritoshi Fukushima ◽  
Akihiro Hasumi

Zebrafish are easy to breed in a laboratory setting as they are extremely fertile and produce dozens of eggs per set. Because zebrafish eggs and the skin of the early-stage larvae are transparent, their embryos and the hearts and muscles of their larvae can be easily observed. Multiple rapid analyses of heart rate and behavior can be performed on these larvae simultaneously, enabling investigation of the influence of neuroactive substances on abnormal behavior, death, and associated pathogenetic mechanisms. Zebrafish larvae are becoming increasingly popular among researchers and are used in laboratories worldwide to study various vertebrate life phenomena; more experimental systems using zebrafish will undoubtedly be developed in the future. However, based on the available literature, we believe that the conceptualization of a protocol based on scientific evidence is necessary to achieve standardization. We exposed zebrafish larvae at 6–7 days post-fertilization to 50 repeated light–dark stimuli at either 15-min or 5-min intervals. We measured the traveled distance and habituation time through a video tracking apparatus. The traveled distance stabilized after the 16th repetition when the zebrafish were exposed to light–dark stimuli at 15-min intervals and after the 5th repetition when exposed at 5-min intervals. Additionally, at 15-min intervals, the peak of the traveled distance was reached within the first minute in a dark environment, whereas at 5-min intervals, it did not reach the peak even after 5 min. The traveled distance was more stable at 5-min intervals of light/dark stimuli than at 15-min intervals. Therefore, if one acclimatizes zebrafish larvae for 1 h and collects data from the 5th repetition of light/dark stimuli at intervals of 5 min in the light/dark test, a stable traveled distance result can be obtained. The establishment of this standardized method would be beneficial for investigating substances of unknown lethal concentration.

2020 ◽  
Vol 16 (6) ◽  
pp. 774-781
Author(s):  
Liang Wu ◽  
An Kang ◽  
Yujie Lin ◽  
Chenxiao Shan ◽  
Zhu Zhou ◽  
...  

Background: Ilexsaponin A1, one of the most representative triterpene saponin components in the roots of I. pubescens, showed its effects in anticoagulation and antithrombosis, attenuating ischemia-reperfusion-induced myocardial, angiogenesis and inhibiting phosphodiesterase. Objective: Reveal the key intestinal bacterial strains responsible for ilexsaponin A1 metabolism, and clarify their metabolic behavior. Methods: An accurate and sensitive LC-MS/MS method for the determination of “ilexsaponin A1 in General Anaerobic Medium (GAM) broth” was established and systematically validated. Then it was applied to screen and study the metabolic potential of the intestinal bacterial strains in an anaerobic incubation system. Results: Quantitation of ilexsaponin A1 could be performed within an analytical run time of 14.5 min, in the linear range of 2 - 2000 ng/ml. Enterobacter sakazakii, Bifidobacterium breve, Bifidobacterium adolescentis, Bifidobacterium catenulatum, and Bifidobacterium angulatum were identified to have a potential effect to metabolize ilexsaponin A1 to different extents; and further bacterial metabolic studies were performed to clarify their metabolic capacity and behavior. Conclusion: This paper contributes to a better understanding of the intestinal bacterial metabolism of ilexsaponin A1 and provides scientific evidence for its clinical application. Additionally, the importance of intestinal bacterial strains in the disposition of natural products was also highlighted.


2010 ◽  
Vol 107 (5) ◽  
pp. 2043-2047 ◽  
Author(s):  
Zheng Eelderink-Chen ◽  
Gabriella Mazzotta ◽  
Marcel Sturre ◽  
Jasper Bosman ◽  
Till Roenneberg ◽  
...  

Circadian timing is a fundamental biological process, underlying cellular physiology in animals, plants, fungi, and cyanobacteria. Circadian clocks organize gene expression, metabolism, and behavior such that they occur at specific times of day. The biological clocks that orchestrate these daily changes confer a survival advantage and dominate daily behavior, for example, waking us in the morning and helping us to sleep at night. The molecular mechanism of circadian clocks has been sketched out in genetic model systems from prokaryotes to humans, revealing a combination of transcriptional and posttranscriptional pathways, but the clock mechanism is far from solved. Although Saccharomyces cerevisiae is among the most powerful genetic experimental systems and, as such, could greatly contribute to our understanding of cellular timing, it still remains absent from the repertoire of circadian model organisms. Here, we use continuous cultures of yeast, establishing conditions that reveal characteristic clock properties similar to those described in other species. Our results show that metabolism in yeast shows systematic circadian entrainment, responding to cycle length and zeitgeber (stimulus) strength, and a (heavily damped) free running rhythm. Furthermore, the clock is obvious in a standard, haploid, auxotrophic strain, opening the door for rapid progress into cellular clock mechanisms.


2009 ◽  
Vol 15 (1) ◽  
pp. 47-52 ◽  
Author(s):  
Marlena Lembicz ◽  
Paweł Olejniczak ◽  
Ziemowit Olszanowski ◽  
Karolina Górzyńska ◽  
Adrian Leuchtmann

Man-made habitats - hotspots of evolutionary game between grass, fungus and flyThe origin and effects of an evolutionary game between species from three different kingdoms (plants, fungi and animals) are presented. We provide scientific evidence that the interaction discovered in man-made habitats leads to an early stage of coevolution. The grassPuccinellia distanswas observed to rapidly spread in new man-made habitats, while at the same time, it was colonised by the fungusEpichloë typhina.The invasion of infected grasses is accompanied by alterations in life histories of both species:P. distansdeveloped features promoting long-distance spreading, whereasE. typhinachanged its life cycle by forming sexual structures for the second time, later in the vegetative season. This enables the fungus to make use of the late shoots of the grass for sexual reproduction, even though it cannot be completed because the vector of spermatia necessary for fertilisation, femaleBotanophilaflies, is not present at that time. This indicates that such uncoordinated evolutionary processes had taken place before interactions between organisms became so specialised that it is difficult to presume they were the result of natural selection. Moreover, these processes could have been initiated in man-made habitats that, in particular circumstances, can become coevolutionary hotspots.


2017 ◽  
Author(s):  
Jingxuan Xiong

AbstractChlordane is highly toxic organochlorine pesticides that have been widely used throughout the world for decades and posing adverse effects on the environment. Contents detected in tissue and blood samples have resulted in a raising concern for their potential effects on wildlife and humans. In this study, we investigate the potential effect of chlordane on the development of zebrafish embryos. Zebrafish larvae were treated with different concentrations (0, 25, 50, 100, 200 ng/L) of chlordane from 12 hours postfertilization (hpf). Different early stage parameters were observed at 1, 2, 3 and 4 day post-fertilization (dpf). Chlordane-exposed zebrafish larvae appeared significant lower survival rate, developmental and hatching time delay and decreased embryo productivity. The heartbeat rate and blood flow were decreased in a dose dependent manner. These results suggested that exposure to real life of chlordane led to direct morphological and phenotypic changes and effects systems related to development and reproduction even in short-term manner.


2015 ◽  
Vol 35 (suppl_1) ◽  
Author(s):  
Manoj K Bandaru ◽  
Petter Ranefall ◽  
Anastasia Emmanouilidou ◽  
Tiffany Klingström ◽  
Lingjie Tao ◽  
...  

Objectives: Published results show that overfeeding zebrafish larvae on a high-cholesterol diet (HCD) can result in hypercholesterolemia and sub-endothelial lipid deposition in macrophages and other cell types. However, results are so far based on small samples, and the atherogenic response has been heterogeneous. We aim to use zebrafish larvae for large-scale, CRISPR-Cas9-based genetic screens, using results from genome wide association studies for coronary heart disease as a starting point. Firstly however, we need to ensure the model system is appropriate and robust. Therefore, we examined the effect of a high-energy diet (HED) and HCD on vascular lipid deposition in a larger number of larvae (n=241). Methods: Starting at 5 days post fertilization (dpf), ~30 larvae/tank were fed 2x/day on: 1) 5 mg control diet (CD; n=33); 2) 15 mg control diet (HED; n=90); or 3) 15 mg control diet enriched with 4% cholesterol (HCD; n=94). At 14-17 dpf, larvae were soaked in monodansylpentane cadaverase - a lipid staining dye - for 45 min, before imaging the dorsal aorta and caudal vein with a Leica SP5 confocal microscope. We used a custom written script in Cell Profiler to quantify the surface area of lipid deposits in the vasculature. Results: Manual annotation of vascular lipid deposition in 30 images (10 randomly selected images per dietary condition) allowed us to calculate the sensitivity (36%) and specificity (71%) of the Cell Profiler script. Subsequent analyses showed that HED (p=0.004) and HCD (p=0.001) fed larvae have significantly more vascular lipid deposition than CD fed larvae after adjusting for age, batch and vessel length. There was no difference in vascular lipid deposition between HED and HCD fed larvae (p=0.11). Discussion and conclusion: Our results confirm that zebrafish larvae represent a promising model system for early-stage atherosclerosis. In addition, they show that enriching the diet with cholesterol is not required to prompt atherogenesis. Future directions: In the next few months, we will examine if overfeeding also triggers vascular infiltration by macrophages, neutrophils and oxidized LDL cholesterol, and if atherogenesis can be prevented or reduced by treating larvae with statins and/or ezetimibe, using our new, automated imaging setup.


Languages ◽  
2020 ◽  
Vol 5 (4) ◽  
pp. 37
Author(s):  
Yuri E. Vega-Rodríguez ◽  
Elena Garayzabal-Heinze ◽  
Esther Moraleda-Sepúlveda

Prenatal alcohol exposure can cause developmental damage in children. There are different types and ranges of alterations that fall under the name of fetal alcohol spectrum disorders (FASD). Disabilities in learning, cognition, and behavior are observed. Environmental conditions are an influencing factor in this population since they are generally adverse and are either not diagnosed at an early stage or given the appropriate support and approach. We present a case study of a 9-year-old child, in which all the variables affecting his development (FASD diagnosis and socioenvironmental conditions) were observed and analyzed. His early childhood under institutional care, the move to a foster home at the age of 6, and several measures of evaluation from foster care to the present are described. Difficulties in vocabulary, access to vocabulary, morphology, syntax, grammar, oral narrative, pragmatics, speech, and communication were observed, along with cognitive difficulties in memory, perception and executive functioning, social adaptation, learning, and behavior. An early diagnosis and approach enable this population to develop skills in different dimensions to address early adversity despite their neurological and behavioral commitment. Speech-language pathologist services are crucial for the diagnosis and treatment of the language and communication difficulties that characterize this syndrome.


Nanomaterials ◽  
2020 ◽  
Vol 10 (6) ◽  
pp. 1245 ◽  
Author(s):  
Daria Tishkevich ◽  
Sergey Grabchikov ◽  
Tatiana Zubar ◽  
Denis Vasin ◽  
Sergei Trukhanov ◽  
...  

Bi nanocrystalline films were formed from perchlorate electrolyte (PE) on Cu substrate via electrochemical deposition with different duration and current densities. The microstructural, morphological properties, and elemental composition were studied using scanning electron microscopy (SEM), atomic force microscopy (AFM), and energy-dispersive X-ray microanalysis (EDX). The optimal range of current densities for Bi electrodeposition in PE using polarization measurements was demonstrated. For the first time, it was shown and explained why, with a deposition duration of 1 s, co-deposition of Pb and Bi occurs. The correlation between synthesis conditions and chemical composition and microstructure for Bi films was discussed. The analysis of the microstructure evolution revealed the changing mechanism of the films’ growth from pillar-like (for Pb-rich phase) to layered granular form (for Bi) with deposition duration rising. This abnormal behavior is explained by the appearance of a strong Bi growth texture and coalescence effects. The investigations of porosity showed that Bi films have a closely-packed microstructure. The main stages and the growth mechanism of Bi films in the galvanostatic regime in PE with a deposition duration of 1–30 s are proposed.


2016 ◽  
Vol 37 (2) ◽  
pp. 157-176 ◽  
Author(s):  
Laura J. E. Brown ◽  
Tim Adlam ◽  
Faustina Hwang ◽  
Hassan Khadra ◽  
Linda M. Maclean ◽  
...  

The “Novel Assessment of Nutrition and Ageing” (NANA) toolkit is a computerized system for collecting longitudinal information about older adults’ health and behavior. Here, we describe the validation of six items for measuring older adults’ self-reported mood and appetite as part of the NANA system. In Study 1, 48 community-living older adults (aged 65-89 years) completed NANA measures of their current mood and appetite alongside standard paper measures, on three occasions, in a laboratory setting. In Study 2, 40 community-living older adults (aged 64-88 years) completed daily NANA measures of momentary mood and appetite in their own homes, unsupervised, alongside additional measures of health and behavior, over three 7-day periods. The NANA measures were significantly correlated with standard measures of mood and appetite, and showed stability over time. They show utility for tracking mood and appetite longitudinally, and for better understanding links with other aspects of health and behavior.


2020 ◽  
Vol 11 ◽  
Author(s):  
Isabel López-Taboada ◽  
Héctor González-Pardo ◽  
Nélida María Conejo

The Western diet (WD) pattern characterized by high daily intake of saturated fats and refined carbohydrates often leads to obesity and overweight, and it has been linked to cognitive impairment and emotional disorders in both animal models and humans. This dietary pattern alters the composition of gut microbiota, influencing brain function by different mechanisms involving the gut–brain axis. In addition, long-term exposure to highly palatable foods typical of WD could induce addictive-like eating behaviors and hypothalamic-pituitary-adrenal (HPA) axis dysregulation associated with chronic stress, anxiety, and depression. In turn, chronic stress modulates eating behavior, and it could have detrimental effects on different brain regions such as the hippocampus, hypothalamus, amygdala, and several cortical regions. Moreover, obesity and overweight induce neuroinflammation, causing neuronal dysfunction. In this review, we summarize the current scientific evidence about the mechanisms and factors relating WD consumption with altered brain function and behavior. Possible therapeutic interventions and limitations are also discussed, aiming to tackle and prevent this current pandemic.


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