A mouse with an N-ethyl-N-nitrosourea (ENU) induced Trp589Arg Galnt3 mutation represents a model for hyperphosphataemic familial tumoural calcinosis

Esapa, C. T., Head, R. A., Jeyabalan, J., Evans, H., Hough, T. A., Cheeseman, M. T., McNally, E. G., Carr, A. J., Thomas, G. P., Brown, M. A., Croucher, P. I., Brown, S. D. M., Cox, R. D., & Thakker, R. V. (2012) A mouse with an N-ethyl-N-nitrosourea (ENU) induced Trp589Arg Galnt3 mutation represents a model for hyperphosphataemic familial tumoural calcinosis. PLoS ONE, 7(8).

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Mutations of UDP-N-acetyl-alpha-D-galactosamine polypeptide N-acetyl galactosaminyl transferase 3 (GALNT3) result in familial tumoural calcinosis (FTC) and the hyperostosis-hyperphosphataemia syndrome (HHS), which are autosomal recessive disorders characterised by soft-tissue calcification and hyperphosphataemia. To facilitate in vivo studies of these heritable disorders of phosphate homeostasis, we embarked on establishing a mouse model by assessing progeny of mice treated with the chemical mutagen N-ethyl-N-nitrosourea (ENU), and identified a mutant mouse, TCAL, with autosomal recessive inheritance of ectopic calcification, which involved multiple tissues, and hyperphosphataemia; the phenotype was designated TCAL and the locus, Tcal. TCAL males were infertile with loss of Sertoli cells and spermatozoa, and increased testicular apoptosis. Genetic mapping localized Tcal to chromosome 2 (62.64-71.11 Mb) which contained the Galnt3. DNA sequence analysis identified a Galnt3 missense mutation (Trp589Arg) in TCAL mice. Transient transfection of wild-type and mutant Galnt3-enhanced green fluorescent protein (EGFP) constructs in COS-7 cells revealed endoplasmic reticulum retention of the Trp589Arg mutant and Western blot analysis of kidney homogenates demonstrated defective glycosylation of Galnt3 in Tcal/Tcal mice. Tcal/Tcal mice had normal plasma calcium and parathyroid hormone concentrations; decreased alkaline phosphatase activity and intact Fgf23 concentrations; and elevation of circulating 1,25-dihydroxyvitamin D. Quantitative reverse transcriptase-PCR (qRT-PCR) revealed that Tcal/Tcal mice had increased expression of Galnt3 and Fgf23 in bone, but that renal expression of Klotho, 25-hydroxyvitamin D-1α-hydroxylase (Cyp27b1), and the sodium-phosphate co-transporters type-IIa and -IIc was similar to that in wild-type mice. Thus, TCAL mice have the phenotypic features of FTC and HHS, and provide a model for these disorders of phosphate metabolism. © 2012 Esapa et al.

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ID Code: 87612
Item Type: Journal Article
Refereed: Yes
Keywords: 25 hydroxyvitamin D 1alpha hydroxylase, alkaline phosphatase, arginine, calcitriol, calcium, enhanced green fluorescent protein, fibroblast growth factor 23, Klotho protein, oxygenase, parathyroid hormone, tryptophan, unclassified drug, animal cell, animal experiment, animal model, animal tissue, apoptosis, article, bone tissue, calcium blood level, chromosome 2, controlled study, DNA sequence, endoplasmic reticulum, enzyme activity, familial disease, female, gene, gene expression regulation, gene location, gene locus, hyperphosphatemia, in vivo study, kidney homogenate, male, missense mutation, molecular pathology, mouse, mutational analysis, n acetyl galactosaminyl transferase 3 gene, nonhuman, nucleotide sequence, parathyroid hormone blood level, phenotypic variation, phosphate metabolism, progeny, protein expression, Sertoli cell, spermatozoon, transient transfection, tumor calcinosis, vitamin blood level, wild type, Animals, Blotting, Western, Bone and Bones, Calcinosis, Cercopithecus aethiops, COS Cells, Disease Models, Animal, Ethylnitrosourea, Fibroblast Growth Factors, Genes, Recessive, Green Fluorescent Proteins, Hyperostosis, Cortical, Congenital, Mice, Mutation, Missense, N-Acetylgalactosaminyltransferases, Reverse Transcriptase Polymerase Chain Reaction, Sertoli Cells, Spermatozoa, Testis
DOI: 10.1371/journal.pone.0043205
ISSN: 19326203 (ISSN)
Divisions: Current > Schools > School of Biomedical Sciences
Current > Institutes > Institute of Health and Biomedical Innovation
Deposited On: 21 Sep 2015 07:18
Last Modified: 25 Feb 2016 04:17

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