Rapid, high-throughput homogenization of embryonic or larval zebrafish (Danio rerio)

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.

Download Full-text

A zebrafish-centric approach to antiepileptic drug development

Disease Models & Mechanisms ◽

10.1242/dmm.049080 ◽

2021 ◽

Vol 14 (7) ◽

Author(s):

Scott C. Baraban

Keyword(s):

Drug Development ◽

High Throughput ◽

Drug Screening ◽

Danio Rerio ◽

Genetic Manipulation ◽

Dravet Syndrome ◽

Developmental Studies ◽

Compassionate Use ◽

Larval Zebrafish ◽

The Common

ABSTRACT Danio rerio (zebrafish) are a powerful experimental model for genetic and developmental studies. Adaptation of zebrafish to study seizures was initially established using the common convulsant agent pentylenetetrazole (PTZ). Larval PTZ-exposed zebrafish exhibit clear behavioral convulsions and abnormal electrographic activity, reminiscent of interictal and ictal epileptiform discharge. By using this model, our laboratory developed simple locomotion-based and electrophysiological assays to monitor and quantify seizures in larval zebrafish. Zebrafish also offer multiple advantages for rapid genetic manipulation and high-throughput phenotype-based drug screening. Combining these seizure assays with genetically modified zebrafish that represent Dravet syndrome, a rare genetic epilepsy, ultimately contributed to a phenotype-based screen of over 3500 drugs. Several drugs identified in these zebrafish screens are currently in clinical or compassionate-use trials. The emergence of this ‘aquarium-to-bedside’ approach suggests that broader efforts to adapt and improve upon this zebrafish-centric strategy can drive a variety of exciting new discoveries.

Download Full-text

Investigation of NH3 as a selective thyroid hormone receptor modulator in larval zebrafish (Danio rerio)

NeuroToxicology ◽

10.1016/j.neuro.2021.03.003 ◽

2021 ◽

Author(s):

Kyla M. Walter ◽

Latika Singh ◽

Vikrant Singh ◽

Pamela J. Lein

Keyword(s):

Thyroid Hormone ◽

Danio Rerio ◽

Hormone Receptor ◽

Thyroid Hormone Receptor ◽

Larval Zebrafish ◽

Receptor Modulator

Download Full-text

Loss of M2 muscarinic receptor function inhibits development of hypoxic bradycardia and alters cardiac β-adrenergic sensitivity in larval zebrafish (Danio rerio)

AJP Regulatory Integrative and Comparative Physiology ◽

10.1152/ajpregu.00036.2009 ◽

2009 ◽

Vol 297 (2) ◽

pp. R412-R420 ◽

Cited By ~ 17

Author(s):

Shelby L. Steele ◽

Kwok Hong Andy Lo ◽

Vincent Wai Tsun Li ◽

Shuk Han Cheng ◽

Marc Ekker ◽

...

Keyword(s):

Heart Rate ◽

Muscarinic Receptor ◽

Danio Rerio ◽

Antisense Oligonucleotides ◽

Adrenergic Receptor ◽

Larval Zebrafish ◽

M2 Muscarinic Receptor ◽

Β Adrenergic Receptors ◽

Relative Mrna Expression

Fish exposed to hypoxia develop decreased heart rate, or bradycardia, the physiological significance of which remains unknown. The general muscarinic receptor antagonist atropine abolishes the development of this hypoxic bradycardia, suggesting the involvement of muscarinic receptors. In this study, we tested the hypothesis that the hypoxic bradycardia is mediated specifically by stimulation of the M2 muscarinic receptor, the most abundant subtype in the vertebrate heart. Zebrafish ( Danio rerio) were reared at two levels of hypoxia (30 and 40 Torr Po2) from the point of fertilization. In hypoxic fish, the heart rate was significantly lower than in normoxic controls from 2 to 10 days postfertilization (dpf). At the more severe level of hypoxia (30 Torr Po2), there were significant increases in the relative mRNA expression of M 2 and the cardiac type β-adrenergic receptors ( β1AR, β2aAR, and β2bAR) at 4 dpf. The hypoxic bradycardia was abolished (at 40 Torr Po2) or significantly attenuated (at 30 Torr Po2) in larvae experiencing M2 receptor knockdown (using morpholino antisense oligonucleotides). Sham-injected larvae exhibited typical hypoxic bradycardia in both hypoxic regimens. The expression of β1AR, β2aAR, β2bAR, and M 2 mRNA was altered at various stages between 1 and 4 dpf in larvae experiencing M2 receptor knockdown. Interestingly, M2 receptor knockdown revealed a cardioinhibitory role for the β2-adrenergic receptor. This is the first study to demonstrate a specific role of the M2 muscarinic receptor in the initiation of hypoxic bradycardia in fish.

Download Full-text

On the effects of iron ore tailings micro/nanoparticles in embryonic and larval zebrafish (Danio rerio)

The Science of The Total Environment ◽

10.1016/j.scitotenv.2020.143456 ◽

2021 ◽

Vol 759 ◽

pp. 143456

Author(s):

Vítor Otacílio de Almeida ◽

Talita Carneiro Brandão Pereira ◽

Lilian de Souza Teodoro ◽

Manuella Escobar ◽

Carolina Junqueira Ordovás ◽

...

Keyword(s):

Danio Rerio ◽

Iron Ore ◽

Larval Zebrafish ◽

Iron Ore Tailings

Download Full-text

Fumonisin B1 Production by Fusarium Species and Mycotoxigenic Effect on Larval Zebrafish

Tropical Life Sciences Research ◽

10.21315/tlsr2020.31.3.7 ◽

2020 ◽

Vol 31 (3) ◽

pp. 91-107 ◽

Cited By ~ 1

Author(s):

Najihah Azman ◽

Nur Ain Izzati Mohd Zainudin ◽

Wan Norhamidah Wan Ibrahim

Keyword(s):

Liquid Chromatography ◽

Danio Rerio ◽

Morphological Changes ◽

Fusarium Species ◽

Fumonisin B1 ◽

Larval Zebrafish ◽

Zebrafish Larvae ◽

Significant Difference ◽

Successful Amplification ◽

Fast Liquid Chromatography

Fumonisin B1 (FB1) is a common mycotoxin produced by Fusarium species particularly F. proliferatum and F. verticillioides. The toxin produced can cause adverse effects on humans and animals. The objectives of this study were to detect the production of FB1 based on the amplification of FUM1 gene, to quantify FB1 produced by the isolates using Ultra-fast Liquid Chromatography (UFLC) analysis, to examine the embryotoxicity effect of FB1 and to determine EC50 toward the larvae of zebrafish (Danio rerio). Fifty isolates of Fusarium species were isolated from different hosts throughout Malaysia. Successful amplification of the FUM1 gene showed the presence of this gene (800 bp) in the genome of 48 out of 50 isolates. The highest level of FB1 produced by F. proliferatum isolate B2433 was 6677.32 ppm meanwhile F. verticillioides isolate J1363 was 954.01 ppm. From the assessment of embryotoxicity test of FB1 on larvae of zebrafish, five concentrations of FB1 (0.43 ppm, 0.58 ppm, 0.72 ppm, 0.87 ppm and 1.00 ppm) were tested. Morphological changes of the FB1 exposed-larvae were observed at 24 to 168 hpf. The mortality rate and abnormality of zebrafish larvae were significantly increased at 144 hpf exposure. Meanwhile, the spontaneous tail coiling showed a significant difference. There were no significant differences in the heartbeat rate. As a conclusion, the presence of FUM1 in every isolate can be detected by FUM1 gene analysis and both of the species produced different concentrations of FB1. This is the first report of FB1 produced by Fusarium species gave a significant effect on zebrafish development.

Download Full-text