New mouse models for metabolic bone diseases generated by genome-wide ENU mutagenesis

Abstract

Metabolic bone disorders arise as primary diseases or may be secondary due to a multitude of organ malfunctions. Animal models are required to understand the molecular mechanisms responsible for the imbalances of bone metabolism in disturbed bone mineralization diseases. Here we present the isolation of mutant mouse models for metabolic bone diseases by phenotyping blood parameters that target bone turnover within the large-scale genome-wide Munich ENU Mutagenesis Project. A screening panel of three clinical parameters, also commonly used as biochemical markers in patients with metabolic bone diseases, was chosen. Total alkaline phosphatase activity and total calcium and inorganic phosphate levels in plasma samples of F1 offspring produced from ENU-mutagenized C3HeB/FeJ male mice were measured. Screening of 9,540 mice led to the identification of 257 phenodeviants of which 190 were tested by genetic confirmation crosses. Seventy-one new dominant mutant lines showing alterations of at least one of the biochemical parameters of interest were confirmed. Fifteen mutations among three genes (Phex, Casr, and Alpl) have been identified by positional-candidate gene approaches and one mutation of the Asgr1 gene, which was identified by next-generation sequencing. All new mutant mouse lines are offered as a resource for the scientific community.

An ENU Mutagenesis-Derived Mouse Model with a Dominant Jak1 Mutation Resembling Phenotypes of Systemic Autoimmune Disease

“…18 Within this project, we obtained the dominant C3HeB/FeJ-Jak1 S645PMhda (Jak1 S645P ) mouse line carrying a new nonsynonymous point mutation within the codon of a highly conserved serine at position 645 of the pseudokinase domain of the Jak1 gene. Herein, we report the morphological, histological, clinical chemical, and hematological phenotypes we found in Jak1 S645Pþ/À mice.…”

Viable Ednra Y129F mice feature human mandibulofacial dysostosis with alopecia (MFDA) syndrome due to the homologue mutation

“…ENU mutagenesis was performed on the pure inbred C3HeB/FeJ (C3H) mouse strain purchased originally from the Jackson Laboratory (Bar Harbour, Maine) as previously described (Aigner et al 2011; Sabrautzki et al 2012). The mice were housed and handled according to the federal animal welfare guidelines and the responsible authority of Upper Bavaria approved all animal studies (Reference Numbers 55.2-1-54-2532-78-06 and 55.2-1-54-2532-144-10).…”

“…Genotyping was performed by single-nucleotide polymorphism (SNP) analysis using MassExtend (MALDI-TOF MS genotyping system) (Sequenom, San Diego, CA, USA) as previously described (Sabrautzki et al 2012). …”

“…The model was derived by the Munich N -ethyl- N -nitrosourea (ENU) mutagenesis project used for more than a decade as a forward genetic approach to generate mutant mouse models for inherited human diseases (Hrabě de Angelis et al 2000; Sabrautzki et al 2012). Using the standardized phenotyping platform of the German Mouse Clinic (GMC) (Gailus-Durner et al 2009; Fuchs et al 2012), we observed a diversity of dysmorphological, otolaryngeal, and lung phenotypes by comparison of adult heterozygous ( Ednra Y129F/+ ) and homozygous ( Ednra Y129F/Y129F ) mice with their wild-type littermates ( Ednra +/+ ).…”

Genome-Wide ENU Mutagenesis in Combination with High Density SNP Analysis and Exome Sequencing Provides Rapid Identification of Novel Mouse Models of Developmental Disease

“…Once linkage to a chromosomal region is identified candidate gene sequencing can be carried out. If candidates are not evident then recent advancements in next-generation sequencing technologies have made sequencing the exons of hundreds of genes within a chromosomal region feasible [8], [9], [10]. Alternatively to chromosomal linkage analysis whole exome sequencing may be used to identify the causal gene and point mutation [7].…”