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Larval Zebrafish (larval + zebrafish)
Selected AbstractsProtein kinase A modulates A-type potassium currents of larval zebrafish (Danio rerio) white muscle fibresACTA PHYSIOLOGICA, Issue 2 2009C. A. Coutts Abstract Aims:, Potassium (K+) channels are involved in regulating cell excitability and action potential shape. To our knowledge, very little is known about the modulation of A-type K+ currents in skeletal muscle fibres. Therefore, we sought to determine whether K+ currents of zebrafish white skeletal muscle were modulated by protein kinase A (PKA). Methods:, Pharmacology and whole-cell patch clamp were used to examine A-type K+ currents and action potentials associated with zebrafish white skeletal muscle fibres. Results:, Activation of PKA by a combination of forskolin + 3-isobutyl-1-methylxanthine (Fsk + IBMX) decreased the peak current density by ,60% and altered the inactivation kinetics of A-type K+ currents. The specific PKA inhibitor H-89 partially blocked the Fsk + IBMX-induced reduction in peak current density, but had no effect on the change in decay kinetics. Fsk + IBMX treatment did not shift the activation curve, but it significantly reduced the slope factor of activation. Activation of PKA by Fsk + IBMX resulted in a negative shift in the V50 of inactivation. H-89 prevented all Fsk + IBMX-induced changes in the steady-state properties of K+ currents. Application of Fsk + IBMX increased action potential amplitude, but had no significant effect on action potential threshold, half width or recovery rate, when fibres were depolarized with single pulses, paired pulses or with high-frequency stimuli. Conclusion:, PKA modulates the A-type K+ current in zebrafish skeletal muscle and affects action potential properties. Our results provide new insights into the role of A-type K+ channels in muscle physiology. [source] Dual enantioselective effect of the insecticide bifenthrin on locomotor behavior and development in embryonic,larval zebrafishENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 7 2010Meiqing Jin Abstract Bifenthrin (BF) is a synthetic pyrethroid that targets the nervous system of insects and may have adverse effects on the behavior and development of nontarget organisms. However, no reports have been issued on the effects of different enantiomers on locomotor behavior for synthetic pyrethroids (SPs) in zebrafish, and whether locomotor activity is associated with the developmental toxicities remains unclear. In this study, enantioselectivity of BF (1S and 1R) on the acute locomotor activity and developmental toxicities of embryonic,larval zebrafish were first evaluated. The results indicated that 1R -BF was more toxic, causing morphological impairments, with a 96-h median effective concentration (EC50) of 226,µg/L for pericardial edema and 145,µg/L for curved body axis. Administration of 20,µg/L of one enantiomer of BF had differential effects on the locomotor activity of zebrafish larvae at 4 d postfertilization (dpf) under alternating light and dark conditions. Larvae treated with 1R -BF were not sensitive to the alteration of light to dark, and the locomotor activities were reduced to a level similar to that observed in light, which otherwise increased rapidly and markedly. However, 1S -BF did not alter the general pattern of zebrafish response to the light or dark compared with the control. The results demonstrated that the differential effects on development might have contributed to the enantioselectivity in the locomotor activity. The consistency of enantioselectivity with insecticidal activity may also indicate a common mode of action. Furthermore, 1R -BF accelerated the spontaneous movement and hatching process, whereas 1S -BF seemed to be inhibitory. The results suggest the need to link behavioral changes to developmental toxicities in order to achieve more comprehensive health risk assessments of chiral pesticides. Environ. Toxicol. Chem. 2010;29:1561,1567. © 2010 SETAC [source] Dissolved copper triggers cell death in the peripheral mechanosensory system of larval fishENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 2 2006Tiffany L. Linbo Abstract Dissolved copper is an increasingly common non,point source contaminant in urban and urbanizing watersheds. In the present study, we investigated the sublethal effects of dissolved copper on the peripheral mechanosensory system, or lateral line, of larval zebrafish (Danio rerio). Zebrafish larvae were exposed to copper (0,65 ,g/L), and the cytotoxic responses of individual lateral line receptor neurons were examined using a combination of in vivo fluorescence imaging, confocal microscopy, scanning electron microscopy, and conventional histology. Dissolved copper triggered a dose-dependent loss of neurons in identified lateral line neuromasts at concentrations ,20 ,g/L. The onset of cell death in the larval mechanosensory system was rapid (<1 h). When copper-exposed zebrafish were transferred to clean water, the lateral line regenerated over the course of 2 d. In contrast, the lateral line of larvae exposed continuously to dissolved copper (50 ,g/L) for 3 d did not recover. Collectively, these results show that peripheral mechanosensory neurons are vulnerable to the neurotoxic effects of copper. Consequently, dissolved copper in non-point source storm-water runoff has the potential to interfere with rheotaxis, schooling, predator avoidance, and other mechanosensory-mediated behaviors that are important for the migration and survival of fish. [source] In vivo analysis of gut function and disease changes in a zebrafish larvae model of inflammatory bowel disease: A feasibility studyINFLAMMATORY BOWEL DISEASES, Issue 7 2010Angeleen Fleming PhD Abstract Background: The aim of this study was to develop a model of inflammatory bowel disease (IBD) in zebrafish larvae, together with a method for the rapid assessment of gut morphology and function in vivo thereby enabling medium-throughput compound screening. Methods: Assays were performed using larval zebrafish from 3,8 days postfertilization (d.p.f.) in 96-well plates. Gut morphology and peristalsis were observed in vivo using fluorescent imaging following ingestion of fluorescent dyes. IBD was induced by addition of 2,4,6-trinitrobenzenesulfonic acid (TNBS) to the medium within the well. Pathology was assessed in vivo using fluorescent imaging and postmortem by histology, immunohistochemistry, and electron microscopy. Therapeutic compounds were evaluated by coadministration with TNBS. Results: A novel method of investigating gut architecture and peristalsis was devised using fluorescent imaging of live zebrafish larvae. Archetypal changes in gut architecture consistent with colitis were observed throughout the gut. Significant changes in goblet cell number and tumor necrosis factor alpha (TNF-,) antibody staining were used to quantify disease severity and rescue. Prednisolone and 5-amino salicylic acid treatment ameliorated the disease changes. Candidate therapeutic compounds (NOS inhibitors, thalidomide, and parthenolide) were assessed and a dissociation was observed between efficacy assessed using a single biochemical measure (TNF-, staining) versus an assessment of the entire disease state. Conclusions: Gut physiology and pathology relevant to human disease state can be rapidly modeled in zebrafish larvae. The model is suitable for medium-throughput chemical screens and is amenable to genetic manipulation, hence offers a powerful novel premammalian adjunct to the study of gastrointestinal disease. (Inflamm Bowel Dis 2010) [source] Development of zebrafish (Danio rerio) pectoral fin musculatureJOURNAL OF MORPHOLOGY, Issue 2 2005D.H. Thorsen Abstract During posthatching development the fins of fishes undergo striking changes in both structure and function. In this article we examine the development of the pectoral fins from larval through adult life history stages in the zebrafish (Danio rerio), describing in detail their pectoral muscle morphology. We explore the development of muscle structure as a way to interpret the fins' role in locomotion. Genetic approaches in the zebrafish model are providing new tools for examining fin development and we take advantage of transgenic lines in which fluorescent protein is expressed in specific tissues to perform detailed three-dimensional, in vivo fin imaging. The fin musculature of larval zebrafish is organized into two thin sheets of fibers, an abductor and adductor, one on each side of an endoskeletal disk. Through the juvenile stage the number of muscle fibers increases and muscle sheets cleave into distinct muscle subdivisions as fibers orient to the developing fin skeleton. By the end of the juvenile period the pectoral girdle and fin muscles have reoriented to take on the adult organization. We find that this change in morphology is associated with a switch of fin function from activity during axial locomotion in larvae to use in swim initiation and maneuvering in adults. The examination of pectoral fins of the zebrafish highlights the yet to be explored diversity of fin structure and function in subadult developmental stages. J. Morphol. © 2005 Wiley-Liss, Inc. [source] Analysis of gastrointestinal physiology using a novel intestinal transit assay in zebrafishNEUROGASTROENTEROLOGY & MOTILITY, Issue 3 2009H. A. Field Abstract, Gastrointestinal function depends upon coordinated contractions to mix and propel food through the gut. Deregulation of these contractions leads to alterations in the speed of material transit through the gut, with potentially significant consequences. We have developed a method for visualizing intestinal transit, the physiological result of peristaltic contractions, in larval zebrafish. This method allows direct, non-invasive observation of luminal content as it traverses the gut. Using this method, we characterized gastrointestinal transit in zebrafish larvae at 7 days postfertilization. In addition, we used this transit assay to assess the physiological consequences of reduced or absent enteric neurones on intestinal transit in larval zebrafish. This may facilitate the use of the zebrafish for investigating the effect of compounds and candidate genes on gastrointestinal motility. [source] Distribution of prospective glutamatergic, glycinergic, and GABAergic neurons in embryonic and larval zebrafishTHE JOURNAL OF COMPARATIVE NEUROLOGY, Issue 1 2004Shin-Ichi Higashijima Abstract Zebrafish are an excellent model for studies of the functional organization of neuronal circuits, but little is known regarding the transmitter phenotypes of the neurons in their nervous system. We examined the distribution in spinal cord and hindbrain of neurons expressing markers of transmitter phenotype, including the vesicular glutamate transporter (VGLUT) genes for glutamatergic neurons, the neuronal glycine transporter (GLYT2) for glycinergic neurons, and glutamic acid decarboxylase (GAD65/67) for GABAergic neurons. All three markers were expressed in a large domain in the dorsal two-thirds of spinal cord, with additional, more ventral expression domains for VGLUT2 and GAD/GABA. In the large dorsal domain, dual in situ staining showed that GLYT2 -positive cells were intermingled with VGLUT2 cells, with no dual-stained neurons. Many of the neurons in the dorsal expression domain that were positive for GABA markers at embryonic stages were also positive for GLYT2, suggesting that the cells might use both GABA and glycine, at least early in their development. The intermingling of neurons expressing inhibitory and excitatory markers in spinal cord contrasted markedly with the organization in hindbrain, where neurons expressing a particular marker were clustered together to form stripes that were visible running from rostral to caudal in horizontal sections and from dorsomedial to ventrolateral in cross sections. Dual labeling showed that the stripes of neurons labeled with one transmitter marker alternated with stripes of cells labeled for the other transmitter phenotypes. The differences in the distribution of excitatory and inhibitory neurons in spinal cord versus hindbrain may be tied to differences in their patterns of development and functional organization. J. Comp. Neurol. 480:1,18, 2004. © 2004 Wiley-Liss, Inc. [source] Neurotransmitter properties of spinal interneurons in embryonic and larval zebrafishTHE JOURNAL OF COMPARATIVE NEUROLOGY, Issue 1 2004Shin-Ichi Higashijima Abstract Many classes of spinal interneurons in zebrafish have been described based on morphology, but their transmitter phenotypes are largely unknown. Here we combine back-filling or genetic labeling of spinal interneurons with in situ staining for markers of neurotransmitter phenotypes, including the vesicular glutamate transporter (VGLUT) genes for glutamatergic neurons, the neuronal glycine transporter (GLYT2) for glycinergic neurons, and glutamic acid decarboxylase (GAD) for GABAergic neurons. Neurons positive for VGLUT include the commissural CoPA, MCoD, UCoD, and some of the CoSA neurons. The CiD interneurons, which have ipsilateral descending axons, were also VGLUT -positive, as were the ventrally located VeMe interneurons, whose descending axonal trajectory has not been clearly revealed. Cells positive for GLYT2 include the commissural CoLAs as well as some of the CoBL and CoSA neurons. The CiA cells were the only GLYT2 -positive cells with an ipsilateral axon. Cells staining for GAD included, most notably, the dorsal longitudinal ascending (DoLA) and KA interneurons. Our approach allowed us to define the likely transmitter phenotypes of most of the known classes of spinal interneurons. These data provide a foundation for understanding the functional organization of the spinal networks in zebrafish. J. Comp. Neurol. 480:19,37, 2004. © 2004 Wiley-Liss, Inc. [source] | |||||||||||||