Article

Highly Efficient ENU Mutagenesis in Zebrafish.

Hubrecht Institute for Developmental Biology and Stem Cell Research, , Uppsalalaan 8, 3584, CT, Utrecht, The Netherlands.
Methods in Molecular Biology (Impact Factor: 1.29). 02/2009; 546:3-12. DOI: 10.1007/978-1-60327-977-2_1
Source: PubMed

ABSTRACT ENU (N-ethyl-N-nitrosourea) mutagenesis is a widely accepted and proven method to introduce random point mutations in the genome. Because there are no targeted knockout strategies available for zebrafish so far, random mutagenesis is currently the preferred method in both forward and reverse genetic approaches. To obtain high-density mutagenized zebrafish, six consecutive ENU treatments are applied at weekly intervals to adult male zebrafish by bathing them in ENU solution. With this procedure an average germ line mutation load of one mutation every 1.0 x 10(5)-1.5 x 10(5) basepairs is reached routinely in our lab.

0 Bookmarks
 · 
100 Views
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Calcium is of vital importance in vertebrates and plasma levels are tightly regulated. Terrestrial vertebrates depend for their calcium uptake exclusively on the diet, while fish have an essentially infinite directly accessible source of calcium in the surrounding water. The need to store calcium seems thus less evident in fish. It was the goal of the present study to investigate the possible role of the scale compartment of zebrafish (Danio rerio) as an accessible pool for calcium and phosphorus. We restricted the calcium availability to zebrafish both in water and diet. Following 12 days restriction, we found that scalar contents of calcium and phosphorus had declined, while magnesium was unaffected, indicating resorption of calcium hydroxyapatite. Osteoclast activity on scales of calcium-restricted fish increased, as we conclude from enlarged demineralized areas on scales, increased TRAcP activity staining, and higher ctsk gene expression. Osteoblasts respond to calcium limitation by increasing expression of col1a2 and alpl. In the presence of water and dietary calcium, removal of scales from one flank of the fish does not affect scalar mineral contents, scale demineralization and osteoclastic TRAcP activity; expression of sp7, alpl and ctsk are up-regulated. Apparently, a high demand for calcium in zebrafish is preferentially complemented with exogenous calcium, while calcium is recruited from the scales when environmental calcium is limited. We conclude that the scales represent an important deposit of available calcium and phosphorus from which these minerals can be recruited in periods of high demand. This finding contributes to a niche for scales as model in bone research.
    Journal of Applied Ichthyology 08/2014; 30(4). DOI:10.1111/jai.12513 · 0.90 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Neurodegenerative diseases can occur so early as to affect neurodevelopment. From a cohort of more than 2,000 consanguineous families with childhood neurological disease, we identified a founder mutation in four independent pedigrees in cleavage and polyadenylation factor I subunit 1 (CLP1). CLP1 is a multifunctional kinase implicated in tRNA, mRNA, and siRNA maturation. Kinase activity of the CLP1 mutant protein was defective, and the tRNA endonuclease complex (TSEN) was destabilized, resulting in impaired pre-tRNA cleavage. Germline clp1 null zebrafish showed cerebellar neurodegeneration that was rescued by wild-type, but not mutant, human CLP1 expression. Patient-derived induced neurons displayed both depletion of mature tRNAs and accumulation of unspliced pre-tRNAs. Transfection of partially processed tRNA fragments into patient cells exacerbated an oxidative stress-induced reduction in cell survival. Our data link tRNA maturation to neuronal development and neurodegeneration through defective CLP1 function in humans.
    Cell 04/2014; 157(3):651-63. DOI:10.1016/j.cell.2014.03.049 · 31.96 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: Wireless or RF energy transfer will be capable of supplying energy to mobile nodes for packet transmission without physical battery changing or replacement. In this paper, we consider the network in which the access point can transfer wireless energy to the nodes and the nodes use that energy to transmit packets. The nodes can send requests (i.e., bids) to the access point for transferring wireless energy. As the access point adopts auction mechanism for wireless energy transfer, we formulate a noncooperative game to analyze the competitive wireless energy transfer bidding of the nodes. The Nash equilibrium is considered to be a solution of the game. To reach the Nash equilibrium, the nodes as players can adapt their strategies based on stochastic response dynamic. The Markov chain is used to analyze the properties of the game (e.g., convergence). The numerical results clearly show the convergence of the bidding strategy adaptation of the nodes toward the Nash equilibrium.
    ICC 2014 - 2014 IEEE International Conference on Communications; 06/2014

Full-text (2 Sources)

Download
36 Downloads
Available from
Jul 4, 2014