scholarly journals The swimming plus-maze test: a novel high-throughput model for assessment of anxiety-related behaviour in larval zebrafish. (Danio rerio)

2018 ◽  
Author(s):  
Zoltán K Varga ◽  
Áron Zsigmond ◽  
Diána Pejtsik ◽  
Máté Varga ◽  
Kornél Demeter ◽  
...  
Keyword(s):  
High Throughput ◽  
Danio Rerio ◽  
High Throughput Screening ◽  
Larval Fish ◽  
Imaging Techniques ◽  
Experimental Conditions ◽  
Larval Zebrafish ◽  
Throughput Model ◽  
Plus Maze ◽  
Resolution Imaging

AbstractLarval zebrafish (Danio rerio) has the potential to supplement rodent models due to the availability of resource efficient methods implying high-throughput screening and high-resolution imaging techniques. Although behavioural models are available in larvae, only a few, insensitive approaches can be employed to assess anxiety. Here we present the swimming plus-maze (SPM) test paradigm to assess anxiety-related states in young zebrafish. The “+” shaped apparatus consists of arms of different depth representing differentially aversive context. The paradigm was validated i.) in larval and juvenile zebrafish, ii.) after administration of compounds affecting human anxiety and iii.) in differentially aversive experimental conditions. Furthermore, we compared the SPM with conventional “anxiety tests” of larvae such as the open tank and light/dark tank tests to identify their shared characteristics. We clarified that the preference towards deeper water is conserved trough the ontogenesis and can be abolished by anxiolytic or enhanced by anxiogenic agents, respectively. The behavioural read-out is insensitive to the aversiveness of the platform and unrelated to behaviours assessed by conventional tests utilizing larval fish. Taken together, we developed a sensitive high-throughput test measuring anxiety-related responses of larval zebrafish, which likely reflect bottom-dwelling behaviour of adults, potentially supporting larva-based integrative approaches.

scholarly journals The swimming plus-maze test: a novel high-throughput model for assessment of anxiety-related behaviour in larval and juvenile zebrafish (Danio rerio)

2018 ◽  
Vol 8 (1) ◽  
Author(s):  
Zoltán K. Varga ◽  
Áron Zsigmond ◽  
Diána Pejtsik ◽  
Máté Varga ◽  
Kornél Demeter ◽  
...  
Keyword(s):  
High Throughput ◽  
Danio Rerio ◽  
Maze Test ◽  
Throughput Model ◽  
Plus Maze

scholarly journals Automated Sample Preparation and Data Collection Workflow for High-Throughput In Vitro Metabolomics

Metabolites ◽  
2022 ◽  
Vol 12 (1) ◽  
pp. 52
Author(s):  
Julia M. Malinowska ◽  
Taina Palosaari ◽  
Jukka Sund ◽  
Donatella Carpi ◽  
Gavin R. Lloyd ◽  
...  
Keyword(s):  
Sample Preparation ◽  
High Throughput ◽  
High Throughput Screening ◽  
In Vitro Screening ◽  
Chemical Safety ◽  
Experimental Conditions ◽  
Well Location ◽  
Relative Standard ◽  
Automated Sample Preparation

Regulatory bodies have started to recognise the value of in vitro screening and metabolomics as two types of new approach methodologies (NAMs) for chemical risk assessments, yet few high-throughput in vitro toxicometabolomics studies have been reported. A significant challenge is to implement automated sample preparation of the low biomass samples typically used for in vitro screening. Building on previous work, we have developed, characterised and demonstrated an automated sample preparation and analysis workflow for in vitro metabolomics of HepaRG cells in 96-well microplates using a Biomek i7 Hybrid Workstation (Beckman Coulter) and Orbitrap Elite (Thermo Scientific) high-resolution nanoelectrospray direct infusion mass spectrometry (nESI-DIMS), across polar metabolites and lipids. The experimental conditions evaluated included the day of metabolite extraction, order of extraction of samples in 96-well microplates, position of the 96-well microplate on the instrument’s deck and well location within a microplate. By using the median relative standard deviation (mRSD (%)) of spectral features, we have demonstrated good repeatability of the workflow (final mRSD < 30%) with a low percentage of features outside the threshold applied for statistical analysis. To improve the quality of the automated workflow further, small method modifications were made and then applied to a large cohort study (4860 sample infusions across three nESI-DIMS assays), which confirmed very high repeatability of the whole workflow from cell culturing to metabolite measurements, whilst providing a significant improvement in sample throughput. It is envisioned that the automated in vitro metabolomics workflow will help to advance the application of metabolomics (as a part of NAMs) in chemical safety, primarily as an approach for high throughput screening and prioritisation.

scholarly journals High-Throughput Screening for Human Galactokinase Inhibitors

2008 ◽  
Vol 13 (5) ◽  
pp. 415-423 ◽  
Author(s):  
Klaas J. Wierenga ◽  
Kent Lai ◽  
Peter Buchwald ◽  
Manshu Tang
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Expression System ◽  
Chemical Compounds ◽  
Average Concentration ◽  
Effective Therapy ◽  
Experimental Conditions ◽  
Classic Galactosemia ◽  
Neonatal Lethality

Inherited deficiency of galactose-1-phosphate uridyltransferase (GALT) can result in a potentially lethal disorder called classic galactosemia. Although the neonatal lethality associated with this disease can be prevented through early diagnosis and a galactose-restricted diet, the lack of effective therapy continues to have consequences: developmental delay, neurological disorders, and premature ovarian failure are common sequelae in childhood and adulthood. Several lines of evidence indicate that an elevated level of galactose-1-phosphate (gal-1-p), the product of galactokinase (GALK), is a major, if not sole, pathogenic mechanism in patients with classic galactosemia. The authors hypothesize that elimination of gal-1-p production by inhibiting GALK will relieve GALT-deficient cells from galactose toxicity. To test this hypothesis, they obtained human GALK using a bacterial expression system. They developed a robust, miniaturized, high-throughput GALK assay (Z′ factor = 0.91) and used this assay to screen against libraries composed of 50,000 chemical compounds with diverse structural scaffolds. They selected 150 compounds that, at an average concentration of 33.3 µM, inhibited GALK activity in vitro more than 86.5% and with a reproducibility score of at least 0.7 for a confirmatory screen under identical experimental conditions. Of these 150 compounds, 34 were chosen for further characterization. Preliminary results indicated that these 34 compounds have potential to serve as leads to the development of more effective therapy of classic galactosemia. ( Journal of Biomolecular Screening 2008:415-423)

scholarly journals Cytometry of raft and caveola membrane microdomains: From flow and imaging techniques to high throughput screening assays

2008 ◽  
Vol 73A (7) ◽  
pp. 599-614 ◽  
Author(s):  
Endre Kiss ◽  
Péter Nagy ◽  
Andrea Balogh ◽  
János Szöllősi ◽  
János Matkó
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Imaging Techniques ◽  
Membrane Microdomains ◽  
Screening Assays

scholarly journals A Microarray Screening Platform with an Experimental Conditions Gradient Generator for the High-Throughput Synthesis of Micro/Nanosized Calcium Phosphates

2020 ◽  
Vol 21 (11) ◽  
pp. 3939
Author(s):  
Xiaoyu Li ◽  
Zhifeng Shi ◽  
Lei Liu ◽  
Guanglin Zhu ◽  
Jianhua Zhou ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Calcium Phosphates ◽  
Two Dimensions ◽  
Experimental Conditions ◽  
Biomedical Materials ◽  
Biomedical Material ◽  
Gradient Generator ◽  
Microarray Screening ◽  
Screening Platform

Calcium phosphates (CaP) represent an impressive kind of biomedical material due to their excellent biocompatibility, bioactivity, and biodegradability. Their morphology and structure highly influence their properties and applications. Whilst great progress has been made in research on biomedical materials, there is still a need to develop a method that can rapidly synthesize and screen micro/nanosized biomedical materials. Here, we utilized a microarray screening platform that could provide the high-throughput synthesis of biomedical materials and screen the vital reaction conditions. With this screening platform, 9 × 9 sets of parallel experiments could be conducted simultaneously with one- or two-dimensions of key reaction condition gradients. We used this platform to establish a one-dimensional gradient of the pH and citrate concentration and a two-dimensional gradient of both the Ca/P ratio and pH to synthesize CaP particles with various morphologies. This screening platform also shows the potential to be extended to other reaction systems for rapid high-throughput screening.

scholarly journals Pericardial Injection of Kainic Acid Induces a Chronic Epileptic State in Larval Zebrafish

2021 ◽  
Vol 14 ◽  
Author(s):  
Lise Heylen ◽  
Duc-Hung Pham ◽  
Ann-Sofie De Meulemeester ◽  
Éric Samarut ◽  
Adrianna Skiba ◽  
...  
Keyword(s):  
Kainic Acid ◽  
High Throughput ◽  
High Throughput Screening ◽  
Video Recording ◽  
Field Potential ◽  
Latency Period ◽  
Pharmacological Characterization ◽  
Larval Zebrafish ◽  
Zebrafish Larvae ◽  
Human Pathology

Epilepsy is a common disorder of the brain characterized by spontaneous recurrent seizures, which develop gradually during a process called epileptogenesis. The mechanistic processes underlying the changes of brain tissue and networks toward increased seizure susceptibility are not fully understood. In rodents, injection of kainic acid (KA) ultimately leads to the development of spontaneous epileptic seizures, reflecting similar neuropathological characteristics as seen in patients with temporal lobe epilepsy (TLE). Although this model has significantly contributed to increased knowledge of epileptogenesis, it is technically demanding, costly to operate and hence not suitable for high-throughput screening of anti-epileptic drugs (AEDs). Zebrafish, a vertebrate with complementary advantages to rodents, is an established animal model for epilepsy research. Here, we generated a novel KA-induced epilepsy model in zebrafish larvae that we functionally and pharmacologically validated. KA was administered by pericardial injection at an early zebrafish larval stage. The epileptic phenotype induced was examined by quantification of seizure-like behavior using automated video recording, and of epileptiform brain activity measured via local field potential (LFP) recordings. We also assessed GFP-labeled GABAergic and RFP-labeled glutamatergic neurons in double transgenic KA-injected zebrafish larvae, and examined the GABA and glutamate levels in the larval heads by liquid chromatography with tandem mass spectrometry detection (LC-MS/MS). Finally, KA-injected larvae were exposed to five commonly used AEDs by immersion for pharmacological characterization of the model. Shortly after injection, KA induced a massive damage and inflammation in the zebrafish brain and seizure-like locomotor behavior. An abnormal reorganization of brain circuits was observed, a decrease in both GABAergic and glutamatergic neuronal population and their associated neurotransmitters. Importantly, these changes were accompanied by spontaneous and continuous epileptiform brain discharges starting after a short latency period, as seen in KA rodent models and reminiscent of human pathology. Three out of five AEDs tested rescued LFP abnormalities but did not affect the seizure-like behavior. Taken together, for the first time we describe a chemically-induced larval zebrafish epilepsy model offering unique insights into studying epileptogenic processes in vivo and suitable for high-throughput AED screening purposes and rapid genetic investigations.

scholarly journals In vivo recording of adult zebrafish electrocardiogram and assessment of drug-induced QT prolongation

2006 ◽  
Vol 291 (1) ◽  
pp. H269-H273 ◽  
Author(s):  
David J. Milan ◽  
Ian L. Jones ◽  
Patrick T. Ellinor ◽  
Calum A. MacRae
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Larval Fish ◽  
Low Noise ◽  
Qt Prolongation ◽  
Motion Artifacts ◽  
Adult Zebrafish ◽  
Perfusion System ◽  
Drug Induced

In the last decade the zebrafish has become a major model organism for the study of development and organogenesis. To maximize the experimental utility of this organism, it will be important to establish methods for adult phenotyping. We previously proposed that the embryonic zebrafish may be useful in high-throughput screening for drug-induced cardiotoxicity. We now describe a method for the reproducible recording of the adult zebrafish ECG and illustrate its application in the investigation of QT-prolonging drugs. Zebrafish ECGs were obtained by inserting two needle electrodes through the ventral epidermis. Fish were perfused orally, and motion artifacts were eliminated with a paralytic dose of μ-conotoxin GIIIB. Test compounds were delivered via the perfusion system. Without a means of hydration and oxygenation, the fish succumb rapidly. The use of a perfusion system allowed stable recording for >6 h. Baseline conduction intervals were as follows: PR, 66 ms (SD 14); QRS, 34 ms (SD 11); QT, 242 ms (SD 54); and R-R, 398 ms (SD 77). The known QT-prolonging agents astemizole, haloperidol, pimozide, and terfenadine caused corrected QT increases of 18% (SD 9), 16% (SD 11), 17% (SD 9), and 11% (SD 6), respectively. The control drugs clonidine, penicillin and propranolol did not prolong the corrected QT interval. In conclusion, perfusion and muscular paralysis allows stable, low-noise recording of zebrafish ECGs. Agents known to cause QT prolongation in humans caused QT prolongation in fish in each case. The development of rigorous tools for the phenotyping of adult zebrafish will complement the high-throughput assays currently under development for embryonic and larval fish.

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