Anesthesia and Euthanasia in Zebrafish

Matthews, Monte; Varga, Zoltán

doi:10.1093/ilar.53.2.192

Cited by 241 publications

(166 citation statements)

References 31 publications

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“…Euthanasia was performed with 160 mg/L Tricaine-S pH 7.0 (tricaine methanesulfonate, Syndel, USA) in system water. It should be noted that the relative merits of Tricaine-S and/or hypothermal shock through immersion in ice water (2–4 °C) have been recently debated (Wilson et al, 2009; Matthews and Varga, 2012). While each method has value, current data suggests that hypothermal shock results in a more rapid and humane euthanasia in small tropical species such as zebrafish.…”

Section: Methodsmentioning

confidence: 99%

Comparative analysis of fixation and embedding techniques for optimized histological preparation of zebrafish

Copper

Budgeon

Foutz

et al. 2018

Comparative Biochemistry and Physiology Part C: Toxicology &

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In recognition of the importance of zebrafish as a model organism for studying human disease, we have created zebrafish content for a web-based reference atlas of microanatomy for comparing histology and histopathology between model systems and with humans (http://bio-atlas.psu.edu). Fixation, decalcification, embedding, and sectioning of zebrafish were optimized to maximize section quality. A comparison of protocols involving six fixatives showed that 10% Neutral Buffered Formalin at 21 °C for 24 h yielded excellent results. Sectioning of juveniles and adults requires bone decalcification; EDTA at 0.35 M produced effective decalcification in 21-day-old juveniles through adults (≥~3 Months). To improve section plane consistency in sets of larvae, we have developed new array casting molds based on the outside contours of larvae derived from 3D microCT images. Tissue discontinuity in sections, a common barrier to creating quality sections of zebrafish, was minimized by processing and embedding the formalin-fixed zebrafish tissues in plasticized forms of paraffin wax, and by periodic hydration of the block surface in ice water between sets of sections. Optimal H&E (Hematoxylin and Eosin) staining was achieved through refinement of standard protocols. High quality slide scans produced from glass histology slides were digitally processed to maximize image quality, and experimental replicates posted as full slides as part of this publication. Modifications to tissue processing are still needed to eliminate the need for block surface hydration. The further addition of slide collections from other model systems and 3D tools for visualizing tissue architecture would greatly increase the utility of the digital atlas.

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Section: Methodsmentioning

confidence: 99%

Comparative analysis of fixation and embedding techniques for optimized histological preparation of zebrafish

Copper

Budgeon

Foutz

et al. 2018

Comparative Biochemistry and Physiology Part C: Toxicology &

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“…Fish were individually anesthetized using ethyl 3-aminobenzoate methanesulfonic acid salt (Product #118000500, Acros Organics, Morris Plains, NJ), also known as “MS-222” or “tricaine”, as previously described (Matthews and Varga, 2012). We found it convenient to make Tricaine stock solution (100 ml) in bulk, aliquot it into smaller volumes (10 ml), stored at −20°C, and thaw as needed on a weekly basis.…”

Section: 0 Detailed Methodsmentioning

confidence: 99%

“…Fish, 3 – 15 months old were used for the studies described. Zebrafish lines examined by SLO imaging included Tg(flk1:EGFP) (Jin et al, 2005), Tg(flk1:mCherry), Tg(flk1:nGFP), Tg(l-fabp:DBP-EGFP) (Xie et al, 2010) and Tg(apoeb:lynEGFP)zf147 (Matthews and Varga, 2012; Peri and Nusslein-Volhard, 2008). In the Tg(flk1:EGFP) and Tg(flk1:mCherry) models, EGFP and RFP (mCherry) are expressed in the cytoplasm of vascular endothelial cells while Tg(flk1:nGFP) zebrafish express GFP in the nuclei of vascular endothelial cells.…”

Section: 0 Materials and Suppliesmentioning

confidence: 99%

Retinal vasculature of adult zebrafish: In vivo imaging using confocal scanning laser ophthalmoscopy

Bell

Xie

Yuan

et al. 2014

Experimental Eye Research

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Over the past 3 decades the zebrafish (Danio rerio) has become an important biomedical research species. As their use continues to grow additional techniques and tools will be required to keep pace with ongoing research using this species. In this paper we describe a novel method for in vivo imaging of the retinal vasculature in adult animals using a commercially available confocal scanning laser ophthalmoscope (SLO). With this instrumentation, we demonstrate the ability to distinguish diverse vascular phenotypes in different transgenic GFP lines. In addition this technology allows repeated visualization of the vasculature in individual zebrafish over time to document vascular leakage progression and recovery induced by intraocular delivery of proteins that induce vascular permeability. SLO of the retinal vasculature was found to be highly informative, providing images of high contrast and resolution that were capable of resolving individual vascular endothelial cells. Finally, the procedures required to acquire SLO images from zebrafish are non-invasive, simple to perform and can be achieved with low animal mortality, allowing repeated imaging of individual fish.

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“…In order to obtain brain samples, zebrafish were euthanized by hypothermal shock as described previously and brains were removed by dissection (Matthews and Varga 2012;Wilson et al 2009). Each independent experiment was performed using biological preparations consisted of a ''pool'' of four and six brains for ectonucleotidases and ADA, respectively.…”

Section: Preparation Of Soluble and Membrane Fractionsmentioning

confidence: 99%

Benzodiazepines alter nucleotide and nucleoside hydrolysis in zebrafish (Danio rerio) brain

et al. 2015

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Anxiety is characterized by unpleasant bodily sensations, such as pounding heart and intense fear. The therapy involves the administration of benzodiazepine drugs. Purinergic signaling participates in the induction of several behavioral patterns and their actions are inactivated by ectonucleotidases and adenosine deaminase (ADA). Since there is evidence about the involvement of purinergic system in the actions mediated by benzodiazepines, we evaluated the effects in vitro and in vivo of administration of diazepam and midazolam on nucleoside triphosphate diphosphohydrolases, ecto-5'-nucleotidase, and ADA activities in zebrafish brain, followed by the analysis of gene expression pattern of these enzymes and adenosine receptors (A1, A2a1, A2a2, A2b). The in vitro studies demonstrated that diazepam decreased ATP (66 % for 500 µM) and ADP hydrolysis (40-54 % for 10-500 µM, respectively). Midazolam decreased ATP (16-71 % for 10-500 µM, respectively) and ADP (48-73.5 % for 250-500 µM, respectively) hydrolysis as well as the ecto-ADA activity (26-27.5 % for 10-500 µM, respectively). AMP hydrolysis was decreased in animals treated with of 0.5 and 1 mg/L midazolam (32 and 36 %, respectively). Diazepam and midazolam decreased the ecto-ADA activity at 1.25 mg/L and 1 mg/L (31 and 33 %, respectively), but only 0.1 mg/L midazolam induced an increase (40 %) in cytosolic ADA. The gene expression analysis demonstrated changes on ecto-5'-nucleotidase, A1, A2a1, A2a2, and A2b mRNA transcript levels after acute treatment with benzodiazepines. These findings demonstrated that benzodiazepine exposure induces a modulation of extracellular nucleotide and nucleoside metabolism, suggesting the purinergic signaling may be, at least in part, related to benzodiazepine effects.

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Anesthesia and Euthanasia in Zebrafish

Cited by 241 publications

References 31 publications

Comparative analysis of fixation and embedding techniques for optimized histological preparation of zebrafish

Comparative analysis of fixation and embedding techniques for optimized histological preparation of zebrafish

Retinal vasculature of adult zebrafish: In vivo imaging using confocal scanning laser ophthalmoscopy

Benzodiazepines alter nucleotide and nucleoside hydrolysis in zebrafish (Danio rerio) brain

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