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Johannes Grosse,
Patrick Tarnow,
Holger Römpler,
Boris Schneider,
Reinhard Sedlmeier,
Ulrike Huffstadt, Dirk Korthaus,
Michael Nehls,
Sigrid Wattler,
Torsten Schöneberg,
Heike Biebermann,
Martin Augustin
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ABSTRACT: Chemical random mutagenesis techniques with the germ line supermutagen N-ethyl-N-nitrosourea (ENU) have been established to provide comprehensive collections of mouse models, which were then mined and analyzed in phenotype-driven studies. Here, we applied ENU mutagenesis in a high-throughput fashion for a gene-driven identification of new mutations. Selected members of the large superfamily of G protein-coupled receptors (GPCR), melanocortin type 3 (Mc3r) and type 4 (Mc4r) receptors, and the orphan chemoattractant receptor GPR33, were used as model targets to prove the feasibility of this approach. Parallel archives of DNA and sperm from mice mutagenized with ENU were screened for mutations in these GPCR, and in vitro assays served as a preselection step before in vitro fertilization was performed to generate the appropriate mouse model. For example, mouse models for inherited obesity were established by selecting fully or partially inactivating mutations in Mc4r. Our technology described herein has the potential to provide mouse models for a GPCR dysfunction of choice within <4 mo and can be extended to other gene classes of interest.
Physiological Genomics 08/2006; 26(3):209-17. · 2.73 Impact Factor
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Carola W E Meyer, Dirk Korthaus,
Wolfgang Jagla,
Emmanuelle Cornali,
Johannes Grosse,
Helmut Fuchs,
Martin Klingenspor,
Stephanie Roemheld,
Matthias Tschöp,
Gerhard Heldmaier,
Martin Hrabé De Angelis,
Michael Nehls
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ABSTRACT: The SMA1-mouse is a novel ethyl-nitroso-urea (ENU)-induced mouse mutant that carries an a-->g missense mutation in exon 5 of the GH gene, which translates to a D167G amino acid exchange in the mature protein. Mice carrying the mutation are characterized by dwarfism, predominantly due to the reduction (sma1/+) or absence (sma1/sma1) of the GH-mediated peripubertal growth spurt, with sma1/+ mice displaying a less pronounced phenotype. All genotypes are viable and fertile, and the mode of inheritance is in accordance with a semidominant Mendelian trait. Adult SMA1 mice accumulate excessive amounts of sc and visceral fat in the presence of elevated plasma ghrelin levels, possibly reflecting altered energy partitioning. Our results suggest impaired storage and/or secretion of pituitary GH in mutants, resulting in reduced pituitary GH and reduced GH-stimulated IGF-1 expression. Generation and identification of the SMA1 mouse exemplifies the power of the combination of random mouse mutagenesis with a highly detailed phenotype-analysis as a successful strategy for the detection and analysis of novel gene-function relationships.
Endocrinology 05/2004; 145(5):2531-41. · 4.46 Impact Factor
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Majid Hafezparast,
Rainer Klocke,
Christiana Ruhrberg,
Andreas Marquardt,
Azlina Ahmad-Annuar,
Samantha Bowen,
Giovanna Lalli,
Abi S Witherden,
Holger Hummerich,
Sharon Nicholson, [......],
Martina Rudelius,
Juergen Schlegel,
Helmut Fuchs,
Martin Hrabe de Angelis,
Giampietro Schiavo,
David T Shima,
Andreas P Russ,
Gabriele Stumm,
Joanne E Martin,
Elizabeth M C Fisher
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ABSTRACT: Degenerative disorders of motor neurons include a range of progressive fatal diseases such as amyotrophic lateral sclerosis (ALS), spinal-bulbar muscular atrophy (SBMA), and spinal muscular atrophy (SMA). Although the causative genetic alterations are known for some cases, the molecular basis of many SMA and SBMA-like syndromes and most ALS cases is unknown. Here we show that missense point mutations in the cytoplasmic dynein heavy chain result in progressive motor neuron degeneration in heterozygous mice, and in homozygotes this is accompanied by the formation of Lewy-like inclusion bodies, thus resembling key features of human pathology. These mutations exclusively perturb neuron-specific functions of dynein.
Science 06/2003; 300(5620):808-12. · 31.20 Impact Factor
