Husbandry, General Care, and Transportation of Xenopus laevis and Xenopus tropicalis

McNamara, Sean; Wlizla, Marcin; Horb, Marko E.

doi:10.1007/978-1-4939-8784-9_1

Cited by 37 publications

(29 citation statements)

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“…Additional fluorescence quenching experiments were performed at ambient temperature (20 °C), a situation likely encountered when a frog (i.e. the animal producing and storing these peptides as a cocktail) encounters a bacterial infection in a natural setting 1,2,44,45 . Because the gel-to-liquid crystalline phase transition occurs at about 21-22 °C for the POPE/POPG mixtures studied here, these two phases are expected to coexist at 20 °C 27 .…”

Section: Discussionmentioning

confidence: 99%

Highly synergistic antimicrobial activity of magainin 2 and PGLa peptides is rooted in the formation of supramolecular complexes with lipids

Aisenbrey

Amaro

Pospíšil

et al. 2020

Sci Rep

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Section: Discussionmentioning

confidence: 99%

Highly synergistic antimicrobial activity of magainin 2 and PGLa peptides is rooted in the formation of supramolecular complexes with lipids

Aisenbrey

Amaro

Pospíšil

et al. 2020

Sci Rep

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“…They were maintained in accordance with the established animal housing and procedure protocols at the NXR 77,78 , which were approved by the Institutional Animal Care and Use Committees at OUHSC and MBL. Sexually mature adult male and female Nigerian strain X. tropicalis were purchased from the NXR (RRID:SCR_013731, https ://www.mbl.edu/xenop us), and housed at the OUHSC facility adhering to the protocols described by McNamara et al 77 . Embryos were generated according the procedures described by Wlizla et al 78,79 .…”

Section: Methodsmentioning

confidence: 99%

“…They were transported as 2-week old tadpoles to our satellite aquatics facility at the University of Oklahoma Health Sciences Center (OUHSC). They were maintained in accordance with the established animal housing and procedure protocols at the NXR 77 , 78 , which were approved by the Institutional Animal Care and Use Committees at OUHSC and MBL. Sexually mature adult male and female Nigerian strain X. tropicalis were purchased from the NXR (RRID:SCR_013731, https://www.mbl.edu/xenopus ), and housed at the OUHSC facility adhering to the protocols described by McNamara et al .…”

Section: Methodsmentioning

confidence: 99%

CRISPR/Cas9 mediated mutation of the mtnr1a melatonin receptor gene causes rod photoreceptor degeneration in developing Xenopus tropicalis

Wiechmann

Martin

Horb

2020

Sci Rep

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Nighttime surges in melatonin levels activate melatonin receptors, which synchronize cellular activities with the natural light/dark cycle. Melatonin receptors are expressed in several cell types in the retina, including the photon-sensitive rods and cones. Previous studies suggest that long-term photoreceptor survival and retinal health is in part reliant on melatonin orchestration of circadian homeostatic activities. This scenario would accordingly envisage that disruption of melatonin receptor signaling is detrimental to photoreceptor health. Using in vivo CRISPR/Cas9 genomic editing, we discovered that a small deletion mutation of the Mel1a melatonin receptor (mtnr1a) gene causes a loss of rod photoreceptors in retinas of developing Xenopus tropicalis heterozygous, but not homozygous mutant tadpoles. Cones were relatively spared from degeneration, and the rod loss phenotype was not obvious after metamorphosis. Localization of Mel1a receptor protein appeared to be about the same in wild type and mutant retinas, suggesting that the mutant protein is expressed at some level in mutant retinal cells. The severe impact on early rod photoreceptor viability may signify a previously underestimated critical role in circadian influences on long-term retinal health and preservation of sight. These data offer evidence that disturbance of homeostatic, circadian signaling, conveyed through a mutated melatonin receptor, is incompatible with rod photoreceptor survival. Circadian (daily) rhythms synchronize to the natural solar cycle via photoreceptive cells in the retina of the eye that detect radiant light energy and convey that information to inner retinal cells, some of which project to the brain. Cyclic oscillations of intracellular activity occur in many cells of the body, and are especially robust in retinal cells since they need to anticipate the 12 magnitudes in ambient light intensity changes that can occur over a 24-h period 1. The key circadian signaling molecule of darkness, melatonin, is produced by retinal photoreceptors at nighttime to drive circadian events in the eye and is secreted into the cerebrospinal fluid and general circulation from the pineal gland 2. Melatonin binds to integral membrane G protein-coupled receptors (GPCRs) expressed in target cells throughout the body to harmonize cellular activities with the predictable 24-h day/night cycle 3. Melatonin receptor subtypes are designated as Mel1a, Mel1b, and Mel1c in amphibians, fish and birds 4 , and as MT1, MT2, and GPR50 (which does not bind melatonin) in mammals, respectively 3. One primary function of melatonin in the retina is to enhance visual sensitivity to light at nighttime 5. If the solar light cycle is disengaged from circadian cellular readiness, the metabolic imbalance imposed on darkadapted photoreceptors by radiant light energy causes cell damage 6. The increased vulnerability of photoreceptors to light damage during subjective night 7 is consistent with reports that melatonin receptor activation exacerbates light-induced rod cell death...

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“…2). For instance, aquatic rack systems are commonly used to rear freshwater zebrafish and Xenopus frogs [17][18][19] and widely used for commercial aquaculture [20,21]. These systems typically incorporate mechanical and biological filtration and some percentage of daily water changes to remove accumulated waste products [22].…”

Section: Methods/results/discussionmentioning

confidence: 99%

“…f View of the IR sensor that detects the presence and absence of food in the Teflon feeding line (see Additional files 9,10,11,12). g View of the DC motor controller for the protein skimmer and a small microcontroller unit that regulates power to the skimmer (see Additional files 15,16,17,18). h High magnification view of the Tunze DC motor control board showing the points where 30-gauge wire connections are made to the microcontroller (see Additional file 15).…”

Section: Automated Feedingmentioning

confidence: 99%

An automated aquatic rack system for rearing marine invertebrates

2020

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Background: One hundred years ago, marine organisms were the dominant systems for the study of developmental biology. The challenges in rearing these organisms outside of a marine setting ultimately contributed to a shift towards work on a smaller number of so-called model systems. Those animals are typically non-marine organisms with advantages afforded by short life cycles, high fecundity, and relative ease in laboratory culture. However, a full understanding of biodiversity, evolution, and anthropogenic effects on biological systems requires a broader survey of development in the animal kingdom. To this day, marine organisms remain relatively understudied, particularly the members of the Lophotrochozoa (Spiralia), which include well over one third of the metazoan phyla (such as the annelids, mollusks, flatworms) and exhibit a tremendous diversity of body plans and developmental modes. To facilitate studies of this group, we have previously described the development and culture of one lophotrochozoan representative, the slipper snail Crepidula atrasolea, which is easy to rear in recirculating marine aquaria. Lab-based culture and rearing of larger populations of animals remain a general challenge for many marine organisms, particularly for inland laboratories.Results: Here, we describe the development of an automated marine aquatic rack system for the high-density culture of marine species, which is particularly well suited for rearing filter-feeding animals. Based on existing freshwater recirculating aquatic rack systems, our system is specific to the needs of marine organisms and incorporates robust filtration measures to eliminate wastes, reducing the need for regular water changes. In addition, this system incorporates sensors and associated equipment for automated assessment and adjustment of water quality. An automated feeding system permits precise delivery of liquid food (e.g., phytoplankton) throughout the day, mimicking real-life feeding conditions that contribute to increased growth rates and fecundity. Conclusion: This automated system makes laboratory culture of marine animals feasible for both large and small research groups, significantly reducing the time, labor, and overall costs needed to rear these organisms.

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Husbandry, General Care, and Transportation of Xenopus laevis and Xenopus tropicalis

Cited by 37 publications

References 10 publications

Highly synergistic antimicrobial activity of magainin 2 and PGLa peptides is rooted in the formation of supramolecular complexes with lipids

Highly synergistic antimicrobial activity of magainin 2 and PGLa peptides is rooted in the formation of supramolecular complexes with lipids

CRISPR/Cas9 mediated mutation of the mtnr1a melatonin receptor gene causes rod photoreceptor degeneration in developing Xenopus tropicalis

An automated aquatic rack system for rearing marine invertebrates

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