6 research outputs found

    Genetische und phänotypische Charakterisierung eines Mausmodells für erbliche Polycythämie

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    Im Rahmen des Münchener ENU-Mausmutagenese-Projekts wurde durch chemische Mutagenese die dominante mutante Mauslinie MVD013 erzeugt. Die Linie wurde an Hand einer erhöhten Zahl und eines verringerten Volumens der Erythrozyten etabliert. Außerdem zeigten einzelne ältere mutante Tiere dieser Linie zusätzliche pathologische Veränderungen wie Darmtumoren und Megakolon. Ziel der Arbeit war es, Untersuchungen zur Identifizierung der ursächlichen Mutation in der Mauslinie MVD013 durchzuführen und die pathologischen Auswirkungen zu charakterisieren, um so ihre Eignung als Tiermodell für Polycythämie und Tumorerkrankungen zu evaluieren. Mittels der durchgeführten Grob- und Feinkartierungen wurde die Lage der ursächlichen Mutation auf Chromosom 5, 59,3-89,7 Mb eingegrenzt. Durch Sequenzanalyse der Kandidatengene Kit, Pdgfra und Kdr konnte keine Mutation im kodierenden Bereich dieser Gene nachgewiesen werden. Die mutante Mauslinie MVD013 wurde im Jahr 2008 im Primärscreen der Deutsche Mausklinik untersucht. Zusammen mit den zusätzlichen sekundären Untersuchungen wurde eine umfassende Phänotypbeschreibung der Mauslinie MVD013 erstellt. Die hämatologische Analyse zeigt ein der humanen Polycythämia vera entsprechendes Blutbild, mit leichter Abweichung vom typischen PV Befund in Richtung Thrombozytopenie. Die klinisch-chemischen Untersuchungen ergaben deutlich erniedrigte Glukosespiegel und Eisenkonzentrationen im Plasma der mutanten Tiere. Die Ergebnisse der Analyse zusätzlicher Parameter des Eisenstoffwechsels und die Ergebnissen des intraperitonealen Glukosetoleranztests deuten auf einen erhöhten Eisen- und Glukoseverbrauch bei mutanten Tieren hin. Anhand der Analyse der Blutgasparameter und der Erythropoetinkonzentration im Serum wurde das Vorliegen einer sekundären Erythrozytose ausgeschlossen. Die Untersuchungen der hämatopoetischen Vorläuferzellen geben einen Hinweis auf myelosuppressive Prozesse im Knochenmark der gealterten mutanten Tiere, was mit Symptomen der humanen Post-Polycythämia vera Myelofibrosis vergleichbar ist. Die mutanten Tiere der Linie MVD013 entwickeln gastrointestinale Stromatumoren in Magen und Darm sowie eine Dilatation (Megaceacum) im Bereich des Blinddarms. Somit kann die dominant mutante Mauslinie MVD013 als ein vielversprechendes Mausmodell für Polycythämia vera, gastrointestinale Stromatumoren und pathologische Veränderungen mit Störungen der Darmperistaltik dienen.MVD013, a mutant mouse line showing a dominantly inherited polycytemia phenotype in the peripheral blood cell count, was established within the Munich ENU mouse mutagenesis project by the Clinical Chemistry Screen. Additionally, gastrointestinal tumors and the development of a Megaceacum were observed in individual aged mutant mice. The aim of this project was to characterizing the mutant phenotype of this mouse line and to conduct investigations to identify the causative mutation, in order to evaluate the usefulness of MVD013 mice as a model of polycythemia and other types of cancer. SNP analysis and additional fine mapping showed the highest possibility of the mutation to be located on chromosome 5 between 59,3 and 89,7 Mb. We sequenced the most promising candidate genes Kdr, Kit and Pdgfra. The latter two genes are associated with gastro-intestinal stromal tumors (GIST) and myeloproliferative disorders. Kdr coding for the vascular endothelial growth factor receptor is essential for the differentiation of hematopoietic precursors and is associated with several kinds of cancers. Our results show that these three genes do not have a mutation in the coding part of the gene and in promoter regions of the Kit or Pdgfra gene. Based on a comprehensive phenotypic characterization of this mouse line in a primary screen conducted 2008 by the German Mouse Clinic, and a variety of additional investigation, a comprehensive description of the mutant phenotype has been generated. Concerning the haematological parameters MVD013 mutant mice show similar symptoms like human patients suffering from Polycythemia vera (PV), with the exception of a platelet count tending towards thrombocytopenia, what is usually not seen in human PV-patients. The results obtained from clinical-chemical plasma analysis indicated decreased plasma glucose and iron levels in mutant animals. Together with the data of the IpGTT and additional measures of further iron-metabolism-related parameters, these results indicate an increased glucose and iron consumption in mutant MVD013 mice. A secondary erythrocytosis due to Erythropoetin-stimulation was excluded by additional analyses such as measurement of erythropoietin level in plasma and blood gas analysis. Further the results suggest an increased iron and glucose consumption in mutant mice, and the development of Gastrointestinal stromal tumors (GIST) in association with an impaired intestinal motility leading to Megaceacum-development. This mouse line might represent a model that can be used to elucidate regulatory mechanisms that playing a role in the development of GISTs, peristaltic dysfunction or polycythemias in humans

    Modeling hepatic osteodystrophy in Abcb4 deficient mice.

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    Hepatic osteodystrophy (HOD) denotes the alterations in bone morphology and metabolism frequently observed in patients with chronic liver diseases, in particular in case of cholestatic conditions. The molecular mechanisms underlying HOD are only partially understood. In the present study, we characterized the bone phenotypes of the ATP-binding cassette transporter B4 knockout mouse (Abcb4(-/-)), a well-established mouse model of chronic cholestatic liver disease, with the aim of identifying and characterizing a mouse model for HOD. Furthermore, we investigated the influence of vitamin D on bone quality in this model. The bone morphology analyses revealed reduced bone mineral contents as well as changes in trabecular bone architecture and decreased cortical bone densities in Abcb4(-/-) mice with severe liver fibrosis. We observed dysregulation of genes involved in bone remodeling (osteoprotegerin, osteocalcin, osteopontin) and vitamin D metabolism (7-dehydrocholesterol reductase, Gc-globulin, Cyp2r1, Cyp27a1) as well as alterations in calcium and vitamin D homeostasis. In addition, serum RANKL and TGF-β levels were increased in Abcb4(-/-) mice. Vitamin D dietary intervention was only partially able to restore the bone phenotypes of Abcb4(-/-) animals. We conclude that the Abcb4(-/-) mouse provides an experimental framework and a preclinical model to gain further insights into the molecular pathobiology of HOD and to study the systemic effects of therapeutic interventions. Bone 2013 Aug; 55(2):501-1

    Standardized, systemic phenotypic analysis of Umod(C93F) and Umod(A227T) mutant mice.

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    Uromodulin-associated kidney disease (UAKD) summarizes different clinical features of an autosomal dominant heritable disease syndrome in humans with a proven uromodulin (UMOD) mutation involved. It is often characterized by hyperuricemia, gout, alteration of urine concentrating ability, as well as a variable rate of disease progression inconstantly leading to renal failure and histological alterations of the kidneys. We recently established the two Umod mutant mouse lines Umod(C93F) and Umod(A227T) on the C3H inbred genetic background both showing kidney defects analogous to those found in human UAKD patients. In addition, disease symptoms were revealed that were not yet described in other published mouse models of UAKD. To examine if further organ systems and/or metabolic pathways are affected by Umod mutations as primary or secondary effects, we describe a standardized, systemic phenotypic analysis of the two mutant mouse lines Umod(A227T) and Umod(C93F) in the German Mouse Clinic. Different genotypes as well as different ages were tested. Beside the already published changes in body weight, body composition and bone metabolism, the influence of the Umod mutation on energy metabolism was confirmed. Hematological analysis revealed a moderate microcytic and erythropenic anemia in older Umod mutant mice. Data of the other analyses in 7-10 month-old mutant mice showed single small additional effects

    Innovations in phenotyping of mouse models in the German Mouse Clinic

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    Analysis of mammalian gene function through broad-based phenotypic screens across a consortium of mouse clinics

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