A Dynamic Anesthesia System for Long-Term Imaging in Adult Zebrafish

Wynd, Brenen M.; Watson, Claire J.; Patil, Karuna; Sanders, George E.; Kwon, Ronald Y.

doi:10.1089/zeb.2016.1289

Cited by 11 publications

(7 citation statements)

References 23 publications

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“…Those aggregates on the chorion were also found in NPMA treatments (data not shown). The formation of these aggregates depended on particles sizes, shapes and coating materials that might interact differently with biological molecules such as membrane proteins and DNA [46]. Future works could reveal if the nanoparticles interact with extracellular protein structures, such as chorion proteins.…”

Section: Toxicity Assay For Np and Npma In Zebrafish Embryosmentioning

confidence: 99%

Potential of mucoadhesive nanocapsules in drug release and toxicology in zebrafish

et al. 2020

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Mucoadhesive polymeric nanocapsules have attracted interest of researchers from different fields from natural sciences because of their ability to interact with the mucosa and increase drug permeation. Anesthesia by immersion causes absorption through the skin and gills of fish, so it is important to evaluate the exposure of these organs to drug nanosystems. Benzocaine (BENZ) is one of the most popular anesthetic agents used in fish anesthesia, but it has drawbacks because of its low bioavailability, resulting in weak absorption after immersion. Here we describe method developed for preparing and characterizing chitosan-coated PLGA mucoadhesive nanoparticles containing BENZ (NPMAs) for zebrafish immersion anesthesia. We determined the lowest effective concentration, characterized the interaction of the mucoadhesive system with fish, measured the anesthetic efficacy, and evaluated possible toxic effects in embryos and adults exposed to the nanoformulations. This study opens perspectives for using nanoformulations prepared with BENZ in aquaculture, allowing reduction of dosage as well as promoting more effective anesthesia and improved interaction with the mucoadhesive system of fish.

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Section: Toxicity Assay For Np and Npma In Zebrafish Embryosmentioning

confidence: 99%

Potential of mucoadhesive nanocapsules in drug release and toxicology in zebrafish

et al. 2020

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“…The teleost body surface is covered by scales, bony plates of integumentary skeleton that contain osteoblast monolayers. Here, to track behaviors of scale osteoblasts in vivo, we adapted protocols for long-term anesthesia of adult zebrafish to enable up to 24 hours of continuous live imaging [14, 15]. With new computational tools and a panel of transgenic lines that express fluorescent and photo-switchable reporter genes in osteoblasts, we captured and quantified behaviors of osteoblasts as they form the initial scale pattern.…”

Section: Introductionmentioning

confidence: 99%

In Toto Imaging of Dynamic Osteoblast Behaviors in Regenerating Skeletal Bone

et al. 2018

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SUMMARY Osteoblasts are matrix-depositing cells that can divide and heal bone injuries. Their deep tissue location and the slow progression of bone regeneration challenge attempts to capture osteoblast behaviors in live tissue at high spatiotemporal resolution. Here, we have developed an imaging platform to monitor and quantify individual and collective behaviors of osteoblasts in adult zebrafish scales, skeletal body armor discs that regenerate rapidly after loss. Using a panel of transgenic lines that visualize and manipulate osteoblasts, we find that a founder pool of osteoblasts emerges through de novo differentiation within one day of scale plucking. These osteoblasts undergo division events that are largely uniform in frequency and orientation to establish a primordium. Osteoblast proliferation dynamics diversify across the primordium by two days after injury, with cell divisions focused near, and with orientations parallel to, the scale periphery, occurring coincident with dynamic localization of fgf20a gene expression. In posterior scale regions, cell elongation events initiate in areas soon occupied by mineralized grooves called radii, beginning approximately 2 days postinjury, with patterned osteoblast death events accompanying maturation of these radii. By imaging at single-cell resolution, we detail acquisition of spatiotemporally distinct cell division, motility, and death dynamics within a founder osteoblast pool as bone regenerates.

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“…However, MS-222 induces cardiac arrest in adult zebrafish after exposure as short as 15 minutes, therefore rendering its use limited for long term imaging and longitudinal studies (Huang et al, 2010; Monte & Zoltán, 2012). A combination with either the compound Benzocaine or Isoflurane was found to decrease risks of cardiac arrest and improve recovery (Huang et al 2010; Lockwood et al 2017; Wynd et al 2017). Benzocaine is also known to allow longer staged anaesthesia (Olt et al, 2016).…”

Section: Resultsmentioning

confidence: 99%

Longitudinal brain studies in adult zebrafish by MRI

Hamilton

Allen

Reynolds

2022

Preprint

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Zebrafish (Danio rerio) has been successfully used for decades in developmental studies and disease modelling. The remarkable intake of zebrafish as a model system is partly due to its transparency during the early weeks of its development, allowing in vivo imaging of cellular and molecular processes. However, this key advantage wears off when tissues become opaque as the animal reaches juvenile and adult stages, rendering access to tissues for live imaging and longitudinal studies difficult. Here we aimed to provide a novel approach to image and assess tissue integrity of adult zebrafish using Magnetic Resonance Imaging (MRI) on live zebrafish suitable for longitudinal studies. We built a 3D-printed life support chamber and designed a protocol-directed sedation regime to recover adult zebrafish post scanning in a 9.4T MRI scanner. Our life support chamber is cheap and easy to create using 3D printing, allowing other groups to copy our template for quick setup. Additionally, we optimised the delivery of contrast agent to enhance brain signals in order to refine current delivery, usually delivered by intravenous in rodents. We show here that immersion in gadolinium was a viable alternative to intraperitoneal injection to reduce T1 relaxation times. This could lead to improved image contrast in adult zebrafish disease models. In conclusion, we provide here a detailed methodology to allow longitudinal studies of brain tissue integrity of adult zebrafish, combining safe and efficient delivery of contrast agent and live MRI. This technique can be used to bridge the gap between in vivo studies and longitudinal brain analysis in adult zebrafish which can be applied to the ever-growing number of adult zebrafish models of ageing and neurodegenerative diseases.

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A Dynamic Anesthesia System for Long-Term Imaging in Adult Zebrafish

Cited by 11 publications

References 23 publications

Potential of mucoadhesive nanocapsules in drug release and toxicology in zebrafish

Potential of mucoadhesive nanocapsules in drug release and toxicology in zebrafish

In Toto Imaging of Dynamic Osteoblast Behaviors in Regenerating Skeletal Bone

Longitudinal brain studies in adult zebrafish by MRI

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