Relevant bibliographies by topics / Zebrafish embryos

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Academic literature on the topic 'Zebrafish embryos'

Author: Grafiati
Published: 4 June 2021
Last updated: 25 April 2022

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Journal articles on the topic "Zebrafish embryos"

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Zhu, Zhen, Yangye Geng, Zhangyi Yuan, Siqi Ren, Meijing Liu, Zhaozheng Meng, and Dejing Pan. "A Bubble-Free Microfluidic Device for Easy-to-Operate Immobilization, Culturing and Monitoring of Zebrafish Embryos." Micromachines 10, no. 3 (February 28, 2019): 168. http://dx.doi.org/10.3390/mi10030168.

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The development of miniaturized devices for studying zebrafish embryos has been limited due to complicated fabrication and operation processes. Here, we reported on a microfluidic device that enabled the capture and culture of zebrafish embryos and real-time monitoring of dynamic embryonic development. The device was simply fabricated by bonding two layers of polydimethylsiloxane (PDMS) structures replicated from three-dimensional (3D) printed reusable molds onto a flat glass substrate. Embryos were easily loaded into the device with a pipette, docked in traps by gravity, and then retained in traps with hydrodynamic forces for long-term culturing. A degassing chamber bonded on top was used to remove air bubbles from the embryo-culturing channel and traps so that any embryo movement caused by air bubbles was eliminated during live imaging. Computational fluid dynamics simulations suggested this embryo-trapping and -retention regime to exert low shear stress on the immobilized embryos. Monitoring of the zebrafish embryogenesis over 20 h during the early stages successfully verified the performance of the microfluidic device for culturing the immobilized zebrafish embryos. Therefore, this rapid-prototyping, low-cost and easy-to-operate microfluidic device offers a promising platform for the long-term culturing of immobilized zebrafish embryos under continuous medium perfusion and the high-quality screening of the developmental dynamics.
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Breitwieser, Helmut, Thomas Dickmeis, Marcel Vogt, Marco Ferg, and Christian Pylatiuk. "Fully Automated Pipetting Sorting System for Different Morphological Phenotypes of Zebrafish Embryos." SLAS TECHNOLOGY: Translating Life Sciences Innovation 23, no. 2 (December 8, 2017): 128–33. http://dx.doi.org/10.1177/2472630317745780.

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Systems biology methods, such as transcriptomics and metabolomics, require large numbers of small model organisms, such as zebrafish embryos. Manual separation of mutant embryos from wild-type embryos is a tedious and time-consuming task that is prone to errors, especially if there are variable phenotypes of a mutant. Here we describe a zebrafish embryo sorting system with two cameras and image processing based on template-matching algorithms. In order to evaluate the system, zebrafish rx3 mutants that lack eyes due to a patterning defect in brain development were separated from their wild-type siblings. These mutants show glucocorticoid deficiency due to pituitary defects and serve as a model for human secondary adrenal insufficiencies. We show that the variable phenotypes of the mutant embryos can be safely distinguished from phenotypic wild-type zebrafish embryos and sorted from one petri dish into another petri dish or into a 96-well microtiter plate. On average, classification of a zebrafish embryo takes approximately 1 s, with a sensitivity and specificity of 87% to 95%, respectively. Other morphological phenotypes may be classified and sorted using similar techniques.
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Liu, Lili, Hua Zhu, Yanchun Yan, Peng Lv, and Wei Wu. "Toxicity Evaluation and Biomarker Selection with Validated Reference Gene in Embryonic Zebrafish Exposed to Mitoxantrone." International Journal of Molecular Sciences 19, no. 11 (November 8, 2018): 3516. http://dx.doi.org/10.3390/ijms19113516.

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Notwithstanding the widespread use and promising clinical value of chemotherapy, the pharmacokinetics, toxicology, and mechanism of mitoxantrone remains unclear. To promote the clinical value in the treatment of human diseases and the exploration of potential subtle effects of mitoxantrone, zebrafish embryos were employed to evaluate toxicity with validated reference genes based on independent stability evaluation programs. The most stable and recommended reference gene was gapdh, followed by tubα1b, for the 48 h post fertilization (hpf) zebrafish embryo mitoxantrone test, while both eef1a1l1 and rpl13α were recommended as reference genes for the 96 hpf zebrafish embryo mitoxantrone test. With gapdh as an internal control, we analyzed the mRNA levels of representative hepatotoxicity biomarkers, including fabp10a, gclc, gsr, nqo1, cardiotoxicity biomarker erg, and neurotoxicity biomarker gfap in the 48 hpf embryo mitoxantrone test. The mRNA levels of gclc, gsr, and gfap increased significantly in 10 and 50 μg/L mitoxantrone-treated 48 hpf embryos, while the transcript levels of fabp10a decreased in a dose-dependent manner, indicating that mitoxantrone induced hepatotoxicity and neurotoxicity. Liver hematoxylin–eosin staining and the spontaneous movement of embryos confirmed the results. Thus, the present research suggests that mitoxantrone induces toxicity during the development of the liver and nervous system in zebrafish embryos and that fabp10a is recommended as a potential biomarker for hepatotoxicity in zebrafish embryos. Additionally, gapdh is proposed as a reference gene for the 48 hpf zebrafish embryo mitoxantrone toxicity test, while eef1a1l1 and rpl13α are proposed as that for the 96 hpf test.
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Zhao, Yuliang, Hui Sun, Xiaopeng Sha, Lijia Gu, Zhikun Zhan, and Wen Li. "A Review of Automated Microinjection of Zebrafish Embryos." Micromachines 10, no. 1 (December 24, 2018): 7. http://dx.doi.org/10.3390/mi10010007.

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Cell microinjection is a technique of precise delivery of substances into cells and is widely used for studying cell transfection, signaling pathways, and organelle functions. Microinjection of the embryos of zebrafish, the third most important animal model, has become a very useful technique in bioscience. However, factors such as the small cell size, high cell deformation tendency, and transparent zebrafish embryo membrane make the microinjection process difficult. Furthermore, this process has strict, specific requirements, such as chorion softening, avoiding contacting the first polar body, and high-precision detection. Therefore, highly accurate control and detection platforms are critical for achieving the automated microinjection of zebrafish embryos. This article reviews the latest technologies and methods used in the automated microinjection of zebrafish embryos and provides a detailed description of the current developments and applications of robotic microinjection systems. The review covers key areas related to automated embryo injection, including cell searching and location, cell position and posture adjustment, microscopic visual servoing control, sensors, actuators, puncturing mechanisms, and microinjection.
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Gianoncelli, Alessandra, Michela Guarienti, Martina Fragni, Michela Bertuzzi, Elisa Rossini, Andrea Abate, Ram Manohar Basnet, et al. "Adrenocortical Carcinoma Xenograft in Zebrafish Embryos as a Model To Study the In Vivo Cytotoxicity of Abiraterone Acetate." Endocrinology 160, no. 11 (August 9, 2019): 2620–29. http://dx.doi.org/10.1210/en.2019-00152.

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Abstract Abiraterone acetate (AbiAc) inhibits tumor growth when administered to immunodeficient mice engrafted with the in vitro cell model of human adrenocortical carcinoma (ACC). Here, we developed and validated a zebrafish model engrafted with cortisol-secreting ACC cells to study the effects of AbiAc on tumor growth. The experimental conditions for AbiAc absorption in AB zebrafish embryos including embryo number, AbiAc concentration, and absorption time curve by liquid chromatography–tandem mass spectrometry were set up. The AbiAc effect on steroid production in AB zebrafish embryos was measured as well. ACC cells (the NCI-H295R cell line, the primary cell ACC29, and the negative control cell SW13) were treated with drug-induced liver injury fluorescent dye, and ∼240 cells per 4 nL was injected in the subperidermal space of the yolk sac of AB zebrafish embryos (n = 80 ± 10). The cell area was measured with Noldus DanioScopeTM software. AbiAc absorption in AB zebrafish embryos was stage dependent. Abiraterone (Abi) concentration decreased, whereas its main metabolite, Δ4A, increased. Accordingly, we demonstrated that zebrafish expressed mRNA encoding the enzyme 3β-hydroxysteroid dehydrogenase, which converts Abi in Δ4A. Furthermore, ABiAc reduced cortisol production and increased progesterone in zebrafish embryos. Three days after cell injection, the cortisol-secreting ACC cell area in solvent-treated embryos was significantly higher than that in 1 µM AbiAC‒treated embryos, whereas no AbiAc effect was observed in SW13 cells, which lack the Abi target enzyme CYP17A1.Zebrafish embryos xenografted with ACC tumor cells could be a useful, fast, and reproducible experimental model to preclinically test the activity of new drugs in human ACC.
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Hassan, Sherif M., Eid A. Moussa, and Louise C. Abbott. "Effects of Quillaja Saponin (Quillaja saponaria) on Early Embryonic Zebrafish (Danio rerio) Development." International Journal of Toxicology 27, no. 3 (May 2008): 273–78. http://dx.doi.org/10.1080/10915810802152129.

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Although much attention has focused on environmental contamination by heavy metals, pesticides, and polychlorinated biphenyls, potential deleterious effects of naturally occurring organic compounds have received much less consideration. Saponins, which are glycosides found in many plants, are important, environmentally ubiquitous organic compounds. Saponins have both beneficial and deleterious effects in adults, but little is known about how saponins effect early vertebrate embryonic development. The authors tested the toxicity of quillaja saponin using a zebrafish embryo assay. Quillaja saponin, extracted from bark of the tree, Quillaja saponaria, is a common foaming agent used in foods and beverages. At 6 h post fertilization, zebrafish embryos were exposed to five concentrations (0 [negative control], 1, 5, 10 or 20 μg) of quillaja saponin per milliliter of medium. Zebrafish embryos exposed to 2% ethanol were positive controls (100% embryonic death). Embryos were assessed at 30, 54, and 72 h post fertilization for changes in embryonic development, mortality, time of hatching, and morphological deformities. Embryos exposed to 1 and 5 μg saponin were healthy, showed no obvious deformities, but exhibited shrinkage of the chorion. Hatching time for zebrafish embryos exposed to 1 and 5 μg/ml saponin decreased by 18 h compared to unexposed embryos. Zebrafish embryos treated with 5 μg/ml saponin responded less to touch than embryos treated with 1 μg/ml saponin or controls. Zebrafish embryos exposed to more than 5 μg/ml saponin exhibited 100% embryonic mortality. These results indicate that exposure to 5 μg/ml or less of quillaja saponin acts as a growth promoter, whereas concentrations of 10 μg/ml or greater are lethal.
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Posner, Mason, Kelly L. Murray, Matthew S. McDonald, Hayden Eighinger, Brandon Andrew, Amy Drossman, Zachary Haley, Justin Nussbaum, Larry L. David, and Kirsten J. Lampi. "The zebrafish as a model system for analyzing mammalian and native α-crystallin promoter function." PeerJ 5 (November 27, 2017): e4093. http://dx.doi.org/10.7717/peerj.4093.

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Previous studies have used the zebrafish to investigate the biology of lens crystallin proteins and their roles in development and disease. However, little is known about zebrafish α-crystallin promoter function, how it compares to that of mammals, or whether mammalian α-crystallin promoter activity can be assessed using zebrafish embryos. We injected a variety of α-crystallin promoter fragments from each species combined with the coding sequence for green fluorescent protein (GFP) into zebrafish zygotes to determine the resulting spatiotemporal expression patterns in the developing embryo. We also measured mRNA levels and protein abundance for all three zebrafish α-crystallins. Our data showed that mouse and zebrafish αA-crystallin promoters generated similar GFP expression in the lens, but with earlier onset when using mouse promoters. Expression was also found in notochord and skeletal muscle in a smaller percentage of embryos. Mouse αB-crystallin promoter fragments drove GFP expression primarily in zebrafish skeletal muscle, with less common expression in notochord, lens, heart and in extraocular regions of the eye. A short fragment containing only a lens-specific enhancer region increased lens and notochord GFP expression while decreasing muscle expression, suggesting that the influence of mouse promoter control regions carries over into zebrafish embryos. The two paralogous zebrafish αB-crystallin promoters produced subtly different expression profiles, with the aBa promoter driving expression equally in notochord and skeletal muscle while the αBb promoter resulted primarily in skeletal muscle expression. Messenger RNA for zebrafish αA increased between 1 and 2 days post fertilization (dpf), αBa increased between 4 and 5 dpf, but αBb remained at baseline levels through 5 dpf. Parallel reaction monitoring (PRM) mass spectrometry was used to detect αA, aBa, and αBb peptides in digests of zebrafish embryos. In whole embryos, αA-crystallin was first detected by 2 dpf, peaked in abundance by 4–5 dpf, and was localized to the eye. αBa was detected in whole embryo at nearly constant levels from 1–6 dpf, was also localized primarily to the eye, and its abundance in extraocular tissues decreased from 4–7 dpf. In contrast, due to its low abundance, no αBb protein could be detected in whole embryo, or dissected eye and extraocular tissues. Our results show that mammalian α-crystallin promoters can be efficiently screened in zebrafish embryos and that their controlling regions are well conserved. An ontogenetic shift in zebrafish aBa-crystallin promoter activity provides an interesting system for examining the evolution and control of tissue specificity. Future studies that combine these promoter based approaches with the expanding ability to engineer the zebrafish genome via techniques such as CRISPR/Cas9 will allow the manipulation of protein expression to test hypotheses about lens crystallin function and its relation to lens biology and disease.
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Wijaya, Romel Ciptoadi. "LETHAL CONCENTRATION 50% OF PATCHOULI OIL (Pogostemon cablin) TOWARDS ZEBRAFISH EMBRYO (Danio rerio)." Herb-Medicine Journal 3, no. 2 (August 31, 2020): 1. http://dx.doi.org/10.30595/hmj.v3i2.6360.

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Preface :Patchouli Oil requires toxicity testing for safety before we can use it widely. It causes side effects such as nausea, vomiting and loss of appetite in some people. Determination of lethal concentration 50% (LC50) in the early stages of zebrafish embryos development will provide an easier, faster and precise prediction of toxicity. At a certain dose, it can cause impairment and death toward organisms. Therefore, the aim of this study was to determine LC50 of Patchouli Oil in carboximethyl cellulose emulsifier towards zebrafish embryo (Danio rerio).Method :Laboratory experimental study was using zebrafish embryos at 2 hours post fertilization. The total of 160 embryos were used and divided into 8 groups, i.e.; Negative control (KN) was given the embryonic fluid, positive control 1 was given 5000 ppm of patchouli oil (KP1), positive control was given 5000 ppm of CMC (KP2), and 5 treatment groups, i.e.; concentration of 10 ppm (P1), 30 ppm (P2), 60 ppm (P3), 90 ppm (P4) and 120 ppm (P5) of Patchouli oil which was emulsified in CMC. The study was conducted with 3 times repetition. Data collection was done by calculating total of embryo’s deaths for each treatment at 24-72 hours of exposure. Data were analyzed using Regression Probit Analysis.Results :Mortality in the group of KN was 1.6%, KP1 was 98.3% and KP2 was 0%. Meanwhile in the treatment group of P1, P2, P3, P4 and P5 respectively were 0%, 5%, 10%, 25%, and 50%.Conclusion :Based on these results, Lethal Concentration 50% of Patchouli Oil in CMC emulsifier towards zebrafish embryo is 120 ppm.
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Mendis, Janthri C., Thejani K. Tennakoon, and Chanika D. Jayasinghe. "Zebrafish Embryo Toxicity of a Binary Mixture of Pyrethroid Insecticides: d-Tetramethrin and Cyphenothrin." Journal of Toxicology 2018 (December 26, 2018): 1–8. http://dx.doi.org/10.1155/2018/4182694.

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Pesguard FG161™, a mixture of d-tetramethrin and cyphenothrin (1:3 ratio), is extensively used to achieve rapid control of adult dengue vector, Aedes aegypti, during the disease outbreaks. Both d-tetramethrin and cyphenothrin are synthetic pyrethroids that are known to have adverse effects on non-mammalian organisms such as fish. The present study intended to use zebrafish embryo toxicity model to investigate the toxic effect of the above binary mixture on fish. Particularly, zebrafish embryo toxicity model provides an alternative to acute fish toxicity tests in terms of animal welfare perspective as the embryos are not considered live until 5 days after fertilization. The zebrafish embryos (2 hrs after fertilization) were exposed to a binary mixture of pyrethroids at different concentrations (d-tetramethrin: 0.01 – 1.20 μmolL-1 and cyphenothrin: 0.03 – 3.20 μmolL-1) for 24, 48, and 72 hrs at room temperature (26°C) according to the OECD guideline no. 236. Percentage mortality of embryos were calculated by observing the lethal endpoints and LC50 values were calculated for each time interval employing the probit analysis. This binary mixture was highly toxic to zebrafish embryos and was found to be concentration and time dependent. LC50 values at 24 hrs (d-tet: 0.58 μmolL-1, cyp: 1.74 μmolL-1) were significantly reduced in 48 hrs (d-tet: 0.11 μmolL-1, cyp: 0.33 μmolL-1) and 72 hrs (d-tet: 0.03 μmolL-1, cyp: 0.09 μmolL-1). Coagulation of embryos was the most common lethal effect observed and lack of somite formation and lack of heartbeat were also observed. The present study revealed that the binary mixture is highly toxic to zebrafish embryos even when based on nominal concentrations. Hence, extensive use of these pesticides could be detrimental to fish population and integrated vector control methods which involve the minimum use of insecticides are recommended. Further, this study highlights the applicability of zebrafish embryo toxicity model as an alternative method to investigate the toxicity of pyrethroids to fish.
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Kane, D. A., M. Hammerschmidt, M. C. Mullins, H. M. Maischein, M. Brand, F. J. van Eeden, M. Furutani-Seiki, et al. "The zebrafish epiboly mutants." Development 123, no. 1 (December 1, 1996): 47–55. http://dx.doi.org/10.1242/dev.123.1.47.

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Epiboly, the enveloping of the yolk cell by the blastoderm, is the first zebrafish morphogenetic movement. We isolated four mutations that affect epiboly: half baked, avalanche, lawine and weg. Homozygous mutant embryos arrest the vegetal progress of the deep cells of the blastoderm; only the yolk syncytial layer of the yolk cell and the enveloping layer of the blastoderm reach the vegetal pole of the embryo. The mutations half baked, avalanche and lawine produce a novel dominant effect, termed a zygotic-maternal dominant effect: heterozygous embryos produced from heterozygous females slow down epiboly and accumulate detached cells over the neural tube; a small fraction of these mutant individuals are viable. Heterozygous embryos produced from heterozygous males crossed to homozygous wild-type females complete epiboly normally and are completely viable. Additionally, embryos heterozygous for half baked have an enlarged hatching gland, a partial dominant phenotype. The phenotypes of these mutants demonstrate that, for the spreading of cells during epiboly, the movement of the deep cells of the blastoderm require the function of genes that are not necessary for the movement of the enveloping layer or the yolk cell. Furthermore, the dominant zygotic-maternal effect phenotypes illustrate the maternal and zygotic interplay of genes that orchestrate the early cell movements of the zebrafish.
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Dissertations / Theses on the topic "Zebrafish embryos"

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Ferreira, Rita Joana Rodrigues da Silva Rua. "Cilia motility studies in zebrafish embryos." Master's thesis, Faculdade de Ciências e Tecnologia, 2012. http://hdl.handle.net/10362/7984.

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A thesis submitted in fulfilment of the requirements for the degree of Masters in Molecular Genetics and Biomedicine
Motile ciliary dysfunctions cause specific Ciliopathies that affect mainly the respiratory tract, fertilization and left-right body establishment. The embryonic organ where left-right decisions are first taken is called the organizer, a ciliated organ where a leftward cilia driven fluid-flow is generated. The organizer is named node in the mouse and Kupffer’s vesicle (KV) in zebrafish. The correct left-right axis formation is highly dependent on signaling pathways downstream of such directional fluid-flow. Motile cilia need to be coordinated and Ciliary Beat Frequency (CBF) is characteristic of different types of cilia depending on their function. Using zebrafish as a model, our group has been studying cilia length regulation and motility in wild-type and deltaD-/- mutant embryos. Recently, we showed that Notch signalling was directly involved in the control of cilia length in the KV cells given that the deltaD-/- mutant present shorter KV cilia. The goal of this project was to characterize the CBF of deltaD-/- KV cilia vs. wild-type cilia and reveal how potential differences in CBF impact on KV fluid flow, using spectral analysis associated with highspeed videomicroscopy. By decomposing and comparing the obtained CBF with Fast Fourier Transform, we identified two major populations of motile cilia in wild-type as well as in deltaD-/- mutant embryos. However, we found the CBF populations had differential relative contributions and different distributions between wild-type and mutant embryos. Furthermore, by measuring the velocity of native particles we studied the KV fluid-flow and concluded that the dispersion of the flow velocity was much wider in the deltaD-/- mutants. On the other hand, based on a gene expression study of motility genes downstream of DeltaD, we concluded that motility related genes (dnah7, rsph3 and foxj1a) were deregulated in the mutants. During this project we generated data that led to new hypotheses that will allow us to test the causality between the described correlations.
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Fierro, Jr Javier. "Glutamatergic Synapse Formation in Developing Zebrafish Embryos." Thesis, University of Oregon, 2015. http://hdl.handle.net/1794/18752.

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In order for a human being to process complex thought, cells within the brain must communicate with each other in a very precise manner. The mechanisms which underlie the development of these connections, however, are poorly understood and thus require a thorough investigation. In this dissertation, we attempt to identify components involved in stabilizing synaptic contacts and the mechanisms by which synaptic proteins are trafficked to newly forming contact sites. Interestingly, we also identify a gene involved in the formation of the myotome. To identify proteins involved in stabilizing synaptic contacts, we characterized the function of 4.1B in developing zebrafish embryos. 4.1B is a scaffolding molecule involved in stabilizing protein complexes at sites of cell adhesion. We identified two 4.1B genes in the zebrafish genome, 4.1B-a and 4.1B-b, which are differentially expressed and have evolved divergent functions. 4.1B-a is expressed within the central nervous system, specifically within primary motor neurons. Knockdown studies show a reduction in the number of synapses and altered kinetics of touch evoked-responses, suggesting a role in synaptic stabilization. In contrast, 4.1B-b is primarily expressed in muscle cells. Knockdown of 4.1B-b results in severe muscle fiber disorganization as well as altered locomotor behaviors. Together, these data suggest the basic functions of 4.1B are evolutionarily conserved, with new roles described in the development of synapses and muscle fibers. To determine the mechanisms that underlie protein recruitment to newly forming synapses, we examined the recruitment of three distinct transport packets in the zebrafish spinal cord. During presynaptic assembly, we found synaptic vesicle protein transport vesicles preceded piccolo-containing active zone precursor transport vesicles, which in turn preceded synapsin transport vesicles. We identified the last transport packet as a unique and independent mechanism for the recruitment of synapsin, a protein involved in regulating the reserve pool of synaptic vesicles. Importantly, we found cyclin-dependent kinase 5 regulated the late recruitment of synapsin transport packets to synapses, thus identifying kinases as a key signaling molecule in the formation of synaptic contacts. Together, this work provides new insight into the mechanisms that underlie synaptogenesis. This dissertation includes both previously published and unpublished co-authored material.
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Patterson, Lucy Jane. "Specification of blood and endothelium in zebrafish embryos." Thesis, University of Nottingham, 2005. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.417180.

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Clay, Hilary. "Early host-pathogen interactions during mycobacterial infection of zebrafish embryos /." Thesis, Connect to this title online; UW restricted, 2007. http://hdl.handle.net/1773/5033.

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Ali, Nadeem. "Teratology in zebrafish embryos : a tool for risk assessment /." Uppsala : Dept. of Biomedical Sciences and Veterinary Public Health, Swedish University of Agricultural Sciences, 2007. http://epsilon.slu.se/10665293.pdf.

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Ellingson, Elizabeth A. (Elizabeth Ann) 1979. "Simultaneous visual and electro-cardiogram measurements of zebrafish embryos." Thesis, Massachusetts Institute of Technology, 2001. http://hdl.handle.net/1721.1/36105.

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Thesis (S.B.)--Massachusetts Institute of Technology, Dept. of Mechanical Engineering, 2001.
Includes bibliographical references (p. 37).
An experimental study was performed to determine a simultaneous visual and electrocardiogram measurement of zebrafish embryos. One zebrafish embryo was placed between two electrodes and the electrical signal was amplified 100 times, then a computer recorded the data. The visual reading of the zebrafish heart rate was obtained by viewing the embryo under a microscope. A variety of approaches were investigated to determine the heart rate including amplification, noise filtering and data manipulation. Noise was a significant obstacle in determining the zebrafish embryo's heart rate. Therefore, the signal was smoothed, digitally filtered, and a system transfer function was determined to extract the heart rate from the noisy signal. After the data manipulation, the electrical signal appeared to correspond to the visual reading of the heart rate. Providing a simultaneous visual and electrical measurement of the heart rate can lead to a better understanding of cardiological genetic mutations. This method of measuring the heart rate can supply information on the strength and pattern of the heartbeat, and also detect irregularities in the beat, which could lead to further understanding of cardiological genetic mutations and other related health problems in the future.
by Elizabeth A. Ellingson.
S.B.
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Zhang, Tiantian. "Investigations into the cryopreservation of zebrafish (Brachydanio rerio) embryos." Thesis, University of Bedfordshire, 1994. http://hdl.handle.net/10547/622045.

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Cryopreservation of fish embryos was studied using zebrafish embryo as a model system. Cryoprotectant toxicity, chilling sensitivity and embryo membrane permeability were investigated with a view to developing optimum protocols for cryopreservation of the embryos using controlled slow cooling, non-freezing storage and vitrification. Methanol was found to be the most effective cryoprotectant for controlled slow cooling and non-freezing storage of zebrafish embryos with 11 %heart beat stage embryos surviving after controlled slow cooling to -25°C. Zebrafish embryos were found to be very chilling sensitive with early developmental stages being the most sensitive to chilling injury. Embryo developmental stages after closure of the blastopore (>12-h), especially post heart beat stages were much more resistant to cryoprotectant toxicity and chilling injury. Heart beat stage (27-h) embryos proved to be the best embryo developmental stage for controlled slow cooling and non-freezing storage. Dechorionated embryos are more sensitive to cryoprotectant toxicity and chilling injury. The sensitivity to chilling injury of zebrafish embryos limited the use of controlled slow cooling and non-freezing storage for long term cryopreservation. The attempts at cryopreservation of zebrafish embryos using vitrification produced no embryo survival, although up to 32 % embryos remained morphologically intact immediately after vitrification. Poor cryoprotectant permeation, dehydration and consequently ice formation within the egg are probably the main factors on effecting embryo survival. The results of zebrafish embryo permeability studies demonstrated that the chorion of the embryos was permeable to water and cryoprotectants, whilst the vitelline (plasma) membrane was an effective permeability barrier. The inability to achieve sufficient penetration of the vitelline membrane by cryoprotectants poses severe problems for long term cryopreservation, which need to be overcome, possibly by permeabilisation of the vitelline membrane, before successful cryopreservation can be achieved.
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Gündel, Ulrike. "Proteomics approach for toxicity assessment in Zebrafish (Danio rerio) embryos /." Leipzig : UFZ - Umweltforschungszentrum Leipzig-Halle GmbH in der Helmholtz-Gemeinschaft / Helmholtz Centre for Environmental Research, 2009. http://opac.nebis.ch/cgi-bin/showAbstract.pl?sys=000278302.

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Burkhardt, Markus. "Electron multiplying CCD – based detection in Fluorescence Correlation Spectroscopy and measurements in living zebrafish embryos." Doctoral thesis, Saechsische Landesbibliothek- Staats- und Universitaetsbibliothek Dresden, 2010. http://nbn-resolving.de/urn:nbn:de:bsz:14-qucosa-61021.

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Fluorescence correlation spectroscopy (FCS) is an ultra-sensitive optical technique to investigate the dynamic properties of ensembles of single fluorescent molecules in solution. It is in particular suited for measurements in biological samples. High sensitivity is obtained by employing confocal microscopy setups with diffraction limited small detection volumes, and by using single-photon sensitive detectors, for example avalanche photo diodes (APD). However, fluorescence signal is hence typically collected from a single focus position in the sample only, and several measurements at different positions have to be performed successively. To overcome the time-consuming successive FCS measurements, we introduce electron multiplying CCD (EMCCD) camera-based spatially resolved detection for FCS. With this new detection method, multiplexed FCS measurements become feasible. Towards this goal, we perform FCS measurements with two focal volumes. As an application, we demonstrate spatial cross-correlation measurements between the two detection volumes, which allow to measure calibration-free diffusion coefficients and direction-sensitive processes like molecular flow in microfluidic channels. FCS is furthermore applied to living zebrafish embryos, to investigate the concentration gradient of the morphogen fibroblast growth factor 8 (Fgf8). It is shown by one-focus APD-based and two-focus EMCCD-based FCS, that Fgf8 propagates largely by random diffusion through the extracellular space in developing tissue. The stable concentration gradient is shown to arise from the equilibrium between a local morphogen production and the sink function of the receiving cells by receptor-mediated removal from the extracellular space. The study shows the applicability of FCS to whole model organisms. Especially in such dynamically changing systems in vivo, the perspective of fast parallel FCS measurements is of great importance. In this work, we exemplify parallel, spatially resolved FCS by utilizing an EMCCD camera. The approach, however, can be easily adapted to any other class of two-dimensional array detector. Novel generations of array detectors might become available in the near future, so that multiplexed spatial FCS could then emerge as a standard extension to classical one-focus FCS
Fluoreszenz-Korrelations-Spektroskopie (FCS) ist eine hochempfindliche optische Methode, um die dynamischen Eigenschaften eines Ensembles von einzelnen, fluoreszierenden Molekülen in Lösung zu erforschen. Sie ist insbesondere geeignet für Messungen in biologischen Proben. Die hohe Empfindlichkeit wird erreicht durch Verwendung konfokaler Mikroskop-Aufbauten mit beugungsbegrenztem Detektionsvolumen, und durch Messung der Fluoreszenz mit Einzelphotonen-empfindlichen Detektoren, zum Beispiel Avalanche-Photodioden (APD). Dadurch wird das Fluoreszenzsignal allerdings nur von einer einzelnen Fokusposition in der Probe eingesammelt, und mehrfache Messungen an verschiedenen Positionen in der Probe müssen nacheinander durchgeführt werden. Um die zeitaufwendigen, aufeinanderfolgenden FCS-Einzelmessungen zu überwinden, entwickeln wir in dieser Arbeit Elektronenvervielfachungs-CCD (EMCCD) Kamera-basierte räumlich aufgelöste Detektion für FCS. Mit dieser neuartigen Detektionsmethode werden Multiplex-FCS Messungen möglich. Darauf abzielend führen wir FCS Messungen mit zwei Detektionsvolumina durch. Als Anwendung nutzen wir die räumliche Kreuzkorrelation zwischen dem Signal beider Fokalvolumina. Sie ermöglicht die kalibrationsfreie Bestimmung von Diffusionskoeffizienten und die Messung von gerichteter Bewegung, wie zum Beispiel laminarem Fluss in mikrostrukturierten Kanälen. FCS wird darüber hinaus angewendet auf Messungen in lebenden Zebrafischembryonen, um den Konzentrationsgradienten des Morphogens Fibroblasten-Wachstumsfaktor 8 (Fgf8) zu untersuchen. Mit Hilfe von APD-basierter ein-Fokus FCS und EMCCD-basierter zwei-Fokus FCS zeigen wir, dass Fgf8 hauptsächlich frei diffffundiert im extrazellulären Raum des sich entwickelnden Embryos. Der stabile Konzentrationsgradient entsteht durch ein Gleichgewicht von lokaler Morphogenproduktion und globalem Morphogenabbau durch Rezeptor vermittelte Entfernung aus dem extrazellulären Raum. Die Studie zeigt die Anwendbarkeit von FCS in ganzen Modell-Organismen. Gerade in diesen sich dynamisch ändernden Systemen in vivo ist die Perspektive schneller, paralleler FCS-Messungen von großer Bedeutung. In dieser Arbeit wird räumlich aufgelöste FCS am Beispiel einer EMCCD Kamera durchgeführt. Die Herangehensweise ist jedoch einfach übertragbar auf jede andere Art von zwei-dimensionalem Flächendetektor. Neuartige Flächendetektoren könnten in naher Zukunft verfügbar sein. Dann könnte räumlich aufgelöste Multiplex-FCS eine standardisierte Erweiterung zur klassischen ein-Fokus FCS werden
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Witzel, Sabine. "Local Wnt11 Signalling and its role in coordinating cell behaviour in zebrafish embryos." Doctoral thesis, Saechsische Landesbibliothek- Staats- und Universitaetsbibliothek Dresden, 2006. http://nbn-resolving.de/urn:nbn:de:swb:14-1162424627109-87779.

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Wnt11 is a key signalling molecule that regulates cell polarity/migration during vertebrate development and also promotes the invasive behaviour of adult cancer cells. It is therefore essential to understand the mechanisms by which Wnt11 signalling regulates cell behaviour. The process of vertebrate gastrulation provides an excellent developmental system to study Wnt11 function in vivo. It is known that Wnt11 mediates coordinated cell migration during gastrulation via the non-canonical Wnt pathway that shares several components with a the planar cell polarity pathway (PCP) in Drosophila. However, the mechanisms by which these PCP components facilitate Wnt11 function in vertebrates is still unclear. While in Drosophila, the asymmetric localization of PCP components is crucial for the establishment of cell polarity, no asymmetric localization of Wnt11 pathway components have so far been observed in vertebrates. To shed light on the cellular and molecular mechanisms underlying Wnt11 signalling, I developed an assay to visualize Wnt11 activity in vivo using live imaging of Wnt11 pathway components tagged to fluorescent proteins. This allowed me to determine the sub-cellular distribution of these components and to correlate the effect of Wnt11 activity with the behaviour of living embryonic cells. I found that Wnt11 locally accumulates together with its receptor Frizzled7 (Fz7) at sites of cell-cell contacts and locally recruits the intra-cellular signalling mediator Dishevelled (Dsh) to those sites. Monitoring these apparent Wnt11 signalling centres through time-lapse confocal microscopy revealed, that Wnt11 activity locally increases the persistency of cell-cell contacts. In addition, I found that the atypical cadherin Flamingo (Fmi) is required for this process. Fmi accumulates together with Wnt11/Fz7 at sites of cell-cell contact and locally increased cell adhesion, via a mechanism that appears to be independent of known downstream effectors of Wnt11 signalling such as RhoA and Rok2. This study indicates that Wnt11 locally interacts with Fmi and Fz7 to control cell-contact persistency and to facilitate coherent and coordinated cell migration. This provides a novel mechanism of non-canonical Wnt signalling in mediating cell behaviour, which is likely relevant to other developmental systems. (Die Druckexemplare enthalten jeweils eine CD-ROM als Anlagenteil: 50 MB: Movies - Nutzung: Referat Informationsvermittlung der SLUB)
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Books on the topic "Zebrafish embryos"

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Handley, Heather Martin. Zebrafish cardiovascular cDNA microarrays: Expression profiling and gene discovery in embryos exposed to 2,3,7,8-Tetrachlorodibenzo-P-dioxin. Cambridge, Mass: Massachusetts Institute of Technology, 2003.

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Matthew, Guille, ed. Molecular methods in developmental biology: Xenopus and zebrafish. Totowa, N.J: Humana Press, 1999.

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Guille, Matt. Molecular Methods in Developmental Biology: Xenopus & Zebrafish (Methods in Molecular Biology). Humana Press, 1999.

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Test No. 250: EASZY assay - Detection of Endocrine Active Substances, acting through estrogen receptors, using transgenic tg(cyp19a1b:GFP) Zebrafish embrYos. OECD, 2021. http://dx.doi.org/10.1787/0a39b48b-en.

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Book chapters on the topic "Zebrafish embryos"

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Picker, Alexander, Daniela Roellig, Olivier Pourquié, Andrew C. Oates, and Michael Brand. "Tissue Micromanipulation in Zebrafish Embryos." In Methods in Molecular Biology, 153–72. Totowa, NJ: Humana Press, 2009. http://dx.doi.org/10.1007/978-1-60327-977-2_11.

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Zhong, Hanbing, and Shuo Lin. "Chemical Screening with Zebrafish Embryos." In Methods in Molecular Biology, 193–205. Totowa, NJ: Humana Press, 2011. http://dx.doi.org/10.1007/978-1-61779-012-6_12.

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Perelsman, Ory, Shoh Asano, and Limor Freifeld. "Expansion Microscopy of Larval Zebrafish Brains and Zebrafish Embryos." In Methods in Molecular Biology, 211–22. New York, NY: Springer US, 2022. http://dx.doi.org/10.1007/978-1-0716-2051-9_13.

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Laird, Angela S., and Wim Robberecht. "Modeling Neurodegenerative Diseases in Zebrafish Embryos." In Methods in Molecular Biology, 167–84. Totowa, NJ: Humana Press, 2011. http://dx.doi.org/10.1007/978-1-61779-328-8_11.

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Kamei, Hiroyasu, and Cunming Duan. "Hypoxic Treatment of Zebrafish Embryos and Larvae." In Methods in Molecular Biology, 195–203. New York, NY: Springer New York, 2018. http://dx.doi.org/10.1007/978-1-4939-7665-2_17.

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Yamashita, Michiaki, Kanako Uchino, Yoshimitsu Taguchi, Shintaro Imamura, Daisuke Uchida, Takeshi Yabu, Misako Hojo, and Nobuhiko Ojima. "Stress response and apoptosis in zebrafish embryos." In Aquatic Genomics, 195–206. Tokyo: Springer Japan, 2003. http://dx.doi.org/10.1007/978-4-431-65938-9_17.

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Xu, Qiling, Derek Stemple, and Katherine Joubin. "Microinjection and Cell Transplantation in Zebrafish Embryos." In METHODS IN MOLECULAR BIOLOGY™, 513–20. Totowa, NJ: Humana Press, 2008. http://dx.doi.org/10.1007/978-1-60327-483-8_35.

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Tawk, Marcel, Isaac H. Bianco, and Jonathan D. W. Clarke. "Focal Electroporation in Zebrafish Embryos and Larvae." In Methods in Molecular Biology, 145–51. Totowa, NJ: Humana Press, 2009. http://dx.doi.org/10.1007/978-1-60327-977-2_10.

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Liu, Xingfeng, Qiang Wang, and Anming Meng. "Detection of Smad Signaling in Zebrafish Embryos." In Methods in Molecular Biology, 275–86. New York, NY: Springer New York, 2016. http://dx.doi.org/10.1007/978-1-4939-2966-5_17.

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Gravato, Carlos, Flávia Renata Abe, Danielle Palma de Oliveira, Amadeu M. V. M. Soares, and Inês Domingues. "Acetylcholinesterase (AChE) Activity in Embryos of Zebrafish." In Toxicity Assessment, 119–24. New York, NY: Springer US, 2021. http://dx.doi.org/10.1007/978-1-0716-1091-6_10.

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Conference papers on the topic "Zebrafish embryos"

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Alkawari, Fatima, Wigdan Ali, Fatiha Benslimane, and Huseyin Yalcin. "Investigating the Cardiac effects of New Generation Anti-Diabetic Drug Empagliflozin using Zebrafish Embryo Model." In Qatar University Annual Research Forum & Exhibition. Qatar University Press, 2020. http://dx.doi.org/10.29117/quarfe.2020.0211.

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Type 2 diabetes mellitus (T2DM) affects >16% of adults in Qatar. Newly emerging class of antidiabetic drugs focused on SGLT inhibition were observed to reduce CVDs risks in diabetic patients. Up to date, the mechanism contributing to the CV benefits remains unrevealed. Zebrafish embryos were treated with Aristolochic Acid to induce heart failure then treated with Empagliflozin to determine its beneficial effect. Furthermore the expression of SGLT1 & 2 were determined in the hearts of zebrafish. SGLT2 was expressed more then SGLT1 in the heart and whole embryo. Empa significantly improved the zebrafish embryo'scardiachealthafterinductionofheartfailure.
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Al-Jighefee, Hadeel, Roba Abdin, Gheyath Khalid Nasrallah, and Ola Aljamal. "Toxicity Evaluation of Stearamidopropyl Dimethylamine Surfactant on Embryonic development of Zebrafish." In Qatar University Annual Research Forum & Exhibition. Qatar University Press, 2020. http://dx.doi.org/10.29117/quarfe.2020.0194.

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Background: Surfactants best known for their use in the industry of detergents, household products, and cosmetics. Usually, the amount of released surfactants into the environment is diluted during secondary water treatment. However, there is always a risk of releasing untreated and polluted wastewater containing high amounts of surfactants without knowing the harmful effect on biotic and abiotic elements of the environment. SAPDMA is a surfactant that is used mostly in cosmetics, conditioning agents, and recently in corrosion inhibition. The classification of SAPDMA as a “safe” or “green” surfactant by different environment or chemical agencies is ambiguous, and the literature is lacking. Aim: The aim of this study is to evaluate the safety of SAPDMAusing the zebrafish embryo model. Methods: Zebrafish embryos were exposed to different concentrations of SAPDMA and the effect was assessed by different toxicity assays. This includes mortality/ survival assay to assess the median lethal dose (LC50) teratogenicity assay to assess the no observed effect concentration (NOEC); and organ specific toxicity assays including cardiotoxicity, neurotoxicity (using locomotion assay), and hemoglobin synthesis (using odianisidine staining). Results: Exposure of zebrafish embryos to SAPDMA caused mortality in a dose-dependent manner with a calculated LC50 of 2.3 mg/L. Thus, based on the LC50 value and according to the Fish and Wildlife Service Acute Toxicity Rating Scale, SAPDMA is classified as “moderately toxic”. The NOEC, the concentration that did not cause any significant teratogenicity, was 0.1mg/L. However, this concentration caused significant organ specific and cytotoxic effects, suggesting that harmless concentrations of SAPDMAare lower than 0.1 mg/L. Conclusion: Our data indicate that SAPDMA at very low concentrations causes adverse effects on zebrafish embryos. Thus, we recommend that the use of SAPDMA in industry should be re-evaluated and monitored by the environment and public health agencies.
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Bommer, Kathleen M., Angela DiBenedetto, and Jens O. M. Karlsson. "High-Speed Imaging of Intra-Embryonic Phase Transformation Events During Rapid Freezing of Zebrafish Embryos." In ASME 2011 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2011. http://dx.doi.org/10.1115/sbc2011-53953.

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The zebrafish (Danio rerio) represents an increasingly popular vertebrate animal model valuable for genetic and developmental biology research, due to its rapid rate of reproduction and the ability to directly observe the growing embryos, which are optically clear and develop ex vivo. However, the need to maintain live stock of each genetic strain (the number of which is growing exponentially) is risky and prohibitively costly. Although long-term banking of frozen embryos would solve this problem, to date, no adequate method for cryopreservation of zebrafish embryos has been found (Hagedorn et al., 2004).
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Aires, Ana, Diana Gomes Moreira, Maria de Lourdes Pereira, and Miguel Oliveira. "Effect of cytostatic substances in zebrafish embryos." In 6th International Electronic Conference on Medicinal Chemistry. Basel, Switzerland: MDPI, 2020. http://dx.doi.org/10.3390/ecmc2020-07500.

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Zhao, Lurui, and Eun Sok Kim. "Acoustic Tweezers for Trapping Late-Stage Zebrafish Embryos." In 2019 IEEE 32nd International Conference on Micro Electro Mechanical Systems (MEMS). IEEE, 2019. http://dx.doi.org/10.1109/memsys.2019.8870615.

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Al-Saaidah, Bayan, Waleed Al-Nuaimy, Majid Al-Taee, Iain Young, and Qussay Al-Jubouri. "Identification of tail curvature malformation in zebrafish embryos." In 2017 8th International Conference on Information Technology (ICIT). IEEE, 2017. http://dx.doi.org/10.1109/icitech.2017.8080063.

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Mena, Pamela, Miguel Allende, José Roberto Morales, Ricardo Alarcon, Phil Cole, Andres J. Kreiner, and Hugo F. Arellano. "Survival Study of Zebrafish Embryos Under Gamma Irradiation." In VIII LATIN AMERICAN SYMPOSIUM ON NUCLEAR PHYSICS AND APPLICATIONS. AIP, 2010. http://dx.doi.org/10.1063/1.3480235.

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Xu, Qiaoshu, Tao Di, Xin Zhou, and Ning Gu. "Toxicity Assessment of Silver Nanoparticles using Zebrafish Embryos." In 2017 6th International Conference on Energy and Environmental Protection (ICEEP 2017). Paris, France: Atlantis Press, 2017. http://dx.doi.org/10.2991/iceep-17.2017.85.

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Essa, Asma, Noura Aldous, Fatiha Benslimane, and Huseyin Yalcin. "In Vivo Investigation of Cardiac benefits of Sodium Glucose Cotransporter Inhibition using the Zebrafish Model." In Qatar University Annual Research Forum & Exhibition. Qatar University Press, 2020. http://dx.doi.org/10.29117/quarfe.2020.0199.

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Type 2 diabetes mellitus (T2DM) affects >16% of adults in Qatar. Newly emerging class of antidiabetic drugs, focused on SGLT inhibition were observed to reduce CVDs risks in diabetic patients. Up to date, the mechanism contributing to the CV benefits remains unrevealed. Zebrafish embryos were injected with different morpholinos to knockdown SGLT genes and study their effects on cardiac parameters. SGLT1 inhibition caused the most severe effects on zebrafish embryos with survival rate ~10 %. It also caused tube-like structured heats with edema, affecting significantly the cardiac output and diameter, and increased cardiac markers expressions. Analysis acquired correlates with literature data of SGLT1 predominant expression in heart tissues.
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Lan, Yutao, Baoguo Wang, Xicheng Wang, Tao Wang, Chengxi Wang, Hao Zhang, Jiaojiao Chen, and Wenjie Mei. "Evaluation of tanshinone IIA developmental toxicity in zebrafish embryos." In The 20th International Electronic Conference on Synthetic Organic Chemistry. Basel, Switzerland: MDPI, 2016. http://dx.doi.org/10.3390/ecsoc-20-b019.

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Reports on the topic "Zebrafish embryos"

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Funkenstein, Bruria, and Shaojun (Jim) Du. Interactions Between the GH-IGF axis and Myostatin in Regulating Muscle Growth in Sparus aurata. United States Department of Agriculture, March 2009. http://dx.doi.org/10.32747/2009.7696530.bard.

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Growth rate of cultured fish from hatching to commercial size is a major factor in the success of aquaculture. The normal stimulus for muscle growth in growing fish is not well understood and understanding the regulation of muscle growth in fish is of particular importance for aquaculture. Fish meat constitutes mostly of skeletal muscles and provides high value proteins in most people's diet. Unlike mammals, fish continue to grow throughout their lives, although the size fish attain, as adults, is species specific. Evidence indicates that muscle growth is regulated positively and negatively by a variety of growth and transcription factors that control both muscle cell proliferation and differentiation. In particular, growth hormone (GH), fibroblast growth factors (FGFs), insulin-like growth factors (IGFs) and transforming growth factor-13 (TGF-13) play critical roles in myogenesis during animal growth. An important advance in our understanding of muscle growth was provided by the recent discovery of the crucial functions of myostatin (MSTN) in controlling muscle growth. MSTN is a member of the TGF-13 superfamily and functions as a negative regulator of skeletal muscle growth in mammals. Studies in mammals also provided evidence for possible interactions between GH, IGFs, MSTN and the musclespecific transcription factor My oD with regards to muscle development and growth. The goal of our project was to try to clarify the role of MSTNs in Sparus aurata muscle growth and in particular determine the possible interaction between the GH-IGFaxis and MSTN in regulating muscle growth in fish. The steps to achieve this goal included: i) Determining possible relationship between changes in the expression of growth-related genes, MSTN and MyoD in muscle from slow and fast growing sea bream progeny of full-sib families and that of growth rate; ii) Testing the possible effect of over-expressing GH, IGF-I and IGF-Il on the expression of MSTN and MyoD in skeletal muscle both in vivo and in vitro; iii) Studying the regulation of the two S. aurata MSTN promoters and investigating the possible role of MyoD in this regulation. The major findings of our research can be summarized as follows: 1) Two MSTN promoters (saMSTN-1 and saMSTN-2) were isolated and characterized from S. aurata and were found to direct reporter gene activity in A204 cells. Studies were initiated to decipher the regulation of fish MSTN expression in vitro using the cloned promoters; 2) The gene coding for saMSTN-2 was cloned. Both the promoter and the first intron were found to be polymorphic. The first intron zygosity appears to be associated with growth rate; 3) Full length cDNA coding for S. aurata growth differentiation factor-l I (GDF-II), a closely related growth factor to MSTN, was cloned from S. aurata brain, and the mature peptide (C-terminal) was found to be highly conserved throughout evolution. GDF-II transcript was detected by RT -PCR analysis throughout development in S. aurata embryos and larvae, suggesting that this mRNA is the product of the embryonic genome. Transcripts for GDF-Il were detected by RT-PCR in brain, eye and spleen with highest level found in brain; 4) A novel member of the TGF-Bsuperfamily was partially cloned from S. aurata. It is highly homologous to an unidentified protein (TGF-B-like) from Tetraodon nigroviridisand is expressed in various tissues, including muscle; 5) Recombinant S. aurata GH was produced in bacteria, refolded and purified and was used in in vitro and in vivo experiments. Generally, the results of gene expression in response to GH administration in vivo depended on the nutritional state (starvation or feeding) and the time at which the fish were sacrificed after GH administration. In vitro, recombinantsaGH activated signal transduction in two fish cell lines: RTHI49 and SAFI; 6) A fibroblastic-like cell line from S. aurata (SAF-I) was characterized for its gene expression and was found to be a suitable experimental system for studies on GH-IGF and MSTN interactions; 7) The gene of the muscle-specific transcription factor Myogenin was cloned from S. aurata, its expression and promoter activity were characterized; 8) Three genes important to myofibrillogenesis were cloned from zebrafish: SmyDl, Hsp90al and skNAC. Our data suggests the existence of an interaction between the GH-IGFaxis and MSTN. This project yielded a great number of experimental tools, both DNA constructs and in vitro systems that will enable further studies on the regulation of MSTN expression and on the interactions between members of the GHIGFaxis and MSTN in regulating muscle growth in S. aurata.
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Funkenstein, Bruria, and Cunming Duan. GH-IGF Axis in Sparus aurata: Possible Applications to Genetic Selection. United States Department of Agriculture, November 2000. http://dx.doi.org/10.32747/2000.7580665.bard.

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Many factors affect growth rate in fish: environmental, nutritional, genetics and endogenous (physiological) factors. Endogenous control of growth is very complex and many hormone systems are involved. Nevertheless, it is well accepted that growth hormone (GH) plays a major role in stimulating somatic growth. Although it is now clear that most, if not all, components of the GH-IGF axis exist in fish, we are still far from understanding how fish grow. In our project we used as the experimental system a marine fish, the gilthead sea bream (Sparus aurata), which inhabits lagoons along the Mediterranean and Atlantic coasts of Europe, and represents one of the most important fish species used in the mariculture industry in the Mediterranean region, including Israel. Production of Sparus is rapidly growing, however, in order for this production to stay competitive, the farming of this fish species has to intensify and become more efficient. One drawback, still, in Sparus extensive culture is that it grows relatively slow. In addition, it is now clear that growth and reproduction are physiological interrelated processes that affect each other. In particular sexual maturation (puberty) is known to be closely related to growth rate in fish as it is in mammals, indicating interactions between the somatotropic and gonadotropic axes. The goal of our project was to try to identify the rate-limiting components(s) in Sparus aurata GH-IGF system which might explain its slow growth by studying the ontogeny of growth-related genes: GH, GH receptor, IGF-I, IGF-II, IGF receptor, IGF-binding proteins (IGFBPs) and Pit-1 during early stages of development of Sparus aurata larvae from slow and fast growing lines. Our project was a continuation of a previous BARD project and could be divided into five major parts: i) obtaining additional tools to those obtained in the previous project that are necessary to carry out the developmental study; ii) the developmental expression of growth-related genes and their cellular localization; iii) tissue-specific expression and effect of GH on expression of growth-related genes; iv) possible relationship between GH gene structure, growth rate and genetic selection; v) the possible role of the IGF system in gonadal development. The major findings of our research can be summarized as follows: 1) The cDNAs (complete or partial) coding for Sparus IGFBP-2, GH receptor and Pit-1 were cloned. Sequence comparison reveals that the primary structure of IGFBP-2 protein is 43-49% identical to that of zebrafish and other vertebrates. Intensive efforts resulted in cloning a fragment of 138 nucleotides, coding for 46 amino acids in the proximal end of the intracellular domain of GH receptor. This is the first fish GH receptor cDNA that had been cloned to date. The cloned fragment will enable us to complete the GH - receptor cloning. 2) IGF-I, IGF-II, IGFBP-2, and IGF receptor transcripts were detected by RT-PCR method throughout development in unfertilized eggs, embryos, and larvae suggesting that these mRNAs are products of both the maternal and the embryonic genomes. Preliminary RT-PCR analysis suggest that GH receptor transcript is present in post-hatching larvae already on day 1. 3) IGF-1R transcripts were detected in all tissues tested by RT-PCR with highest levels in gill cartilage, skin, kidney, heart, pyloric caeca, and brain. Northern blot analysis detected IGF receptor only in gonads, brain and gill cartilage but not in muscle; GH increased slightly brain and gill cartilage IGF-1R mRNA levels. 4) IGFBP-2 transcript were detected only in liver and gonads, when analyzed by Northern blots; RT-PCR analysis revealed expression in all tissues studied, with the highest levels found in liver, skin, gonad and pyloric caeca. 5) Expression of IGF-I, IGF-II, IGF-1R and IGFBP-2 was analyzed during gonadal development. High levels of IGF-I and IGFBP-2 expression were found in bisexual young gonads, which decreased during gonadal development. Regardless of maturational stage, IGF-II levels were higher than those of IGF-L 6) The GH gene was cloned and its structure was characterized. It contains minisatellites of tandem repeats in the first and third introns that result in high level of genetic polymorphism. 7) Analysis of the presence of IGF-I and two types of IGF receptor by immunohistochemistry revealed tissue- and stage-specific expression during larval development. Immunohistochemistry also showed that IGF-I and its receptors are present in both testicular and ovarian cells. Although at this stage we are not able to pinpoint which is the rate-limiting step causing the slow growth of Sparus aurata, our project (together with the previous BARD) yielded a great number of experimental tools both DNA probes and antibodies that will enable further studies on the factors regulating growth in Sparus aurata. Our expression studies and cellular localization shed new light on the tissue and developmental expression of growth-related genes in fish.
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