Abstract
Calpainopathy or limb-girdle muscular dystrophy type 2A (LGMD2A) is generally recognized as the most prevalent form of recessive LGMD and is caused by mutations in the CAPN3 gene. Out of a cohort of 119 patients fulfilling clinical criteria for LGMD2, referred to our neuromuscular clinic, 46 were suspected to have LGMD2A, based on western blot results. Four of these patients were shown to have LGMD2I upon molecular analysis, whereas 16 of the remaining 42 patients harbored mutations in CAPN3 by both direct genomic sequencing and cDNA analyses. In 10 patients, we identified both mutant alleles. In three other, only one heterozygous mutation could be identified on the genomic level; however, CAPN3 cDNA analyses demonstrated homozygosity for the mutant allele, indicating the presence of an unidentified allele that somehow compromise correct CAPN3 RNA processing. In the three remaining patients, only a single heterozygous mutation could be identified both at the genomic level and on full-length CAPN3 cDNA. All three patients exhibited a highly abnormal western blot for calpain-3 and clinical characteristics of LGMD2A. Only three of the genetically confirmed LGMD2A patients were of Danish origin, indicating a five- to sixfold lower prevalence in Denmark compared to other European countries. A total of 16 different CAPN3 mutations were identified, of which 5 were novel. The present study demonstrates the value of cDNA analysis for CAPN3 in LGMD2A patients and indicates that calpainopathy is an uncommon cause of LGMD in the Denmark.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
References
Beckmann JS, Brown RH, Muntoni F, Urtizberea A, Bonnemann C, Bushby KM : 66th/67th ENMC sponsored international workshop: the limb-girdle muscular dystrophies, 26–28 March 1999, Naarden, The Netherlands. Neuromuscul Disord 1999; 9: 436–445.
Zatz M, Starling A : Calpains and disease. N Engl J Med 2005; 352: 2413–2423.
Piluso G, Politano L, Aurino S et al: Extensive scanning of the calpain-3 gene broadens the spectrum of LGMD2A phenotypes. J Med Genet 2005; 42: 686–693.
Fanin M, Nascimbeni AC, Fulizio L, Angelini C : The frequency of limb girdle muscular dystrophy 2A in northeastern Italy. Neuromuscul Disord 2005; 15: 218–224.
Pollitt C, Anderson LV, Pogue R, Davison K, Pyle A, Bushby KM : The phenotype of calpainopathy: diagnosis based on a multidisciplinary approach. Neuromuscul Disord 2001; 11: 287–296.
Saenz A, Leturcq F, Cobo AM et al: LGMD2A: genotype-phenotype correlations based on a large mutational survey on the calpain 3 gene. Brain 2005; 128: 732–742.
Richard I, Broux O, Allamand V et al: Mutations in the proteolytic enzyme calpain 3 cause limb-girdle muscular dystrophy type 2A. Cell 1995; 81: 27–40.
Sorimachi H, Toyama-Sorimachi N, Saido TC et al: Muscle-specific calpain, p94, is degraded by autolysis immediately after translation, resulting in disappearance from muscle. J Biol Chem 1993; 268: 10593–10605.
Goll DE, Thompson VF, Li H, Wei W, Cong J : The calpain system. Physiol Rev 2003; 83: 731–801.
Fanin M, Fulizio L, Nascimbeni AC et al: Molecular diagnosis in LGMD2A: mutation analysis or protein testing? Hum Mutat 2004; 24: 52–62.
Anderson LV, Davison K, Moss JA et al: Characterization of monoclonal antibodies to calpain 3 and protein expression in muscle from patients with limb-girdle muscular dystrophy type 2A. Am J Pathol 1998; 153: 1169–1179.
Fanin M, Nardetto L, Nascimbeni AC et al: Correlations between clinical severity, genotype and muscle pathology in limb girdle muscular dystrophy type 2A. J Med Genet 2007; 44: 609–614.
Fanin M, Nascimbeni AC, Fulizio L, Trevisan CP, Meznaric-Petrusa M, Angelini C : Loss of calpain-3 autocatalytic activity in LGMD2A patients with normal protein expression. Am J Pathol 2003; 163: 1929–1936.
The Human Gene Mutation Database HGMD, http://www.hgmd.cf.ac.uk/ac/all.php.
Leiden Muscular Dystrophy Database, http://www.dmd.nl/capn3_seqvar.html.
Balci B, Aurino S, Haliloglu G et al: Calpain-3 mutations in Turkey. Eur J Pediatr 2006; 165: 293–298.
Urtasun M, Saenz A, Roudaut C et al: Limb-girdle muscular dystrophy in Guipuzcoa (Basque Country, Spain). Brain 1998; 121 (Part 9): 1735–1747.
Zatz M, Vainzof M, Passos-Bueno MR : Limb-girdle muscular dystrophy: one gene with different phenotypes, one phenotype with different genes. Curr Opin Neurol 2000; 13: 511–517.
de Paula F, Vainzof M, Passos-Bueno MR et al: Clinical variability in calpainopathy: what makes the difference? Eur J Hum Genet 2002; 10: 825–832.
Richard I, Roudaut C, Saenz A et al: Calpainopathy-a survey of mutations and polymorphisms. Am J Hum Genet 1999; 64: 1524–1540.
Todorova A, Kress W, Mueller C : Novel mutations in the calpain 3 gene in Germany. Clin Genet 2005; 67: 356–358.
Krahn M, Bernard R, Pecheux C et al: Screening of the CAPN3 gene in patients with possible LGMD2A. Clin Genet 2006; 69: 444–449.
Hermanova M, Zapletalova E, Sedlackova J et al: Analysis of histopathologic and molecular pathologic findings in Czech LGMD2A patients. Muscle Nerve 2006; 33: 424–432.
Sveen ML, Schwartz M, Vissing J : High prevalence and phenotype-genotype correlations of limb girdle muscular dystrophy type 2I in Denmark. Ann Neurol 2006; 59: 808–815.
Schwartz M, Hertz JM, Sveen ML, Vissing J : LGMD2I presenting with a characteristic Duchenne or Becker muscular dystrophy phenotype. Neurology 2005; 64: 1635–1637.
Richard I, Brenguier L, Dincer P et al: Multiple independent molecular etiology for limb-girdle muscular dystrophy type 2A patients from various geographical origins. Am J Hum Genet 1997; 60: 1128–1138.
Chrobakova T, Hermanova M, Kroupova I et al: Mutations in Czech LGMD2A patients revealed by analysis of calpain3 mRNA and their phenotypic outcome. Neuromuscul Disord 2004; 14: 659–665.
Deburgrave N, Daoud F, Llense S et al: Protein- and mRNA-based phenotype-genotype correlations in DMD/BMD with point mutations and molecular basis for BMD with nonsense and frameshift mutations in the DMD gene. Hum Mutat 2007; 28: 183–195.
Miller TE, You L, Myerburg RJ, Benke PJ, Bishopric NH : Whole blood RNA offers a rapid, comprehensive approach to genetic diagnosis of cardiovascular diseases. Genet Med 2007; 9: 23–33.
Reese MG, Eeckman FH, Kulp D, Haussler D : Improved splice site detection in Genie. J Comput Biol 1997; 4: 311–323.
ESE finder, http://rulai.cshl.edu/cgi-bin/tools/ESE3/esefinder.cgi.
Stehlikova K, Zapletalova E, Sedlackova J et al: Quantitative analysis of CAPN3 transcripts in LGMD2A patients: involvement of nonsense-mediated mRNA decay. Neuromuscul Disord 2007; 17: 143–147.
Maquat LE : Nonsense-mediated mRNA decay: splicing, translation and mRNP dynamics. Nat Rev Mol Cell Biol 2004; 5: 89–99.
Starling A, Schlesinger D, Kok F, Passos-Bueno MR, Vainzof M, Zatz M : A family with McLeod syndrome and calpainopathy with clinically overlapping diseases. Neurology 2005; 65: 1832–1833.
Minami N, Nishino I, Kobayashi O, Ikezoe K, Goto Y, Nonaka I : Mutations of calpain 3 gene in patients with sporadic limb-girdle muscular dystrophy in Japan. J Neurol Sci 1999; 171: 31–37.
Acknowledgements
Excellent technical assistance was provided by Danuta Goralska-Olsen and Eva Rahtkens.
Author information
Authors and Affiliations
Corresponding author
Additional information
Supplementary Information accompanies the paper on European Journal of Human Genetics website (http://www.nature.com/ejhg)
Supplementary information
Rights and permissions
About this article
Cite this article
Duno, M., Sveen, ML., Schwartz, M. et al. cDNA analyses of CAPN3 enhance mutation detection and reveal a low prevalence of LGMD2A patients in Denmark. Eur J Hum Genet 16, 935–940 (2008). https://doi.org/10.1038/ejhg.2008.47
Received:
Revised:
Accepted:
Published:
Issue date:
DOI: https://doi.org/10.1038/ejhg.2008.47
Keywords
This article is cited by
-
Mutational spectrum of autosomal recessive limb-girdle muscular dystrophies in a cohort of 112 Iranian patients and reporting of a possible founder effect
Orphanet Journal of Rare Diseases (2020)
-
Anoctamin 5 muscular dystrophy in Denmark: prevalence, genotypes, phenotypes, cardiac findings, and muscle protein expression
Journal of Neurology (2013)
-
Calpain 3 is important for muscle regeneration: Evidence from patients with limb girdle muscular dystrophies
BMC Musculoskeletal Disorders (2012)
