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Human Molecular Genetics, 2002, Vol. 11, No. 26 3283-3298
© 2002 Oxford University Press

Gene expression profiling in dysferlinopathies using a dedicated muscle microarray

Stefano Campanaro1, Chiara Romualdi1, Marina Fanin2, Barbara Celegato1, Beniamina Pacchioni1, Silvia Trevisan1, Paolo Laveder1, Cristiano De Pittà1, Elena Pegoraro2, Yukiko K. Hayashi3, Giorgio Valle1, Corrado Angelini2 and Gerolamo Lanfranchi1,*

1CRIBI Biotechnology Centre and Dipartimento di Biologia and 2Dipartimento di Scienze Neurologiche e Psichiatriche, Università degli Studi di Padova, Padova, Italy and 3Department of Neuromuscular Research, National Institute of Neuroscience, Tokyo, Japan

Received July 30, 2002; Accepted October 23, 2002

We have performed expression profiling to define the molecular changes in dysferlinopathy using a novel dedicated microarray platform made with 3'-end skeletal muscle cDNAs. Eight dysferlinopathy patients, defined by western blot, immunohistochemistry and mutation analysis, were investigated with this technology. In a first experiment RNAs from different limb-girdle muscular dystrophy type 2B patients were pooled and compared with normal muscle RNA to characterize the general transcription pattern of this muscular disorder. Then the expression profiles of patients with different clinical traits were independently obtained and hierarchical clustering was applied to discover patient-specific gene variations. MHC class I genes and genes involved in protein biosynthesis were up-regulated in relation to muscle histopathological features. Conversely, the expression of genes codifying the sarcomeric proteins titin, nebulin and telethonin was down-regulated. Neither calpain-3 nor caveolin, a sarcolemmal protein interacting with dysferlin, was consistently reduced. There was a major up-regulation of proteins interacting with calcium, namely S100 calcium-binding proteins and sarcolipin, a sarcoplasmic calcium regulator.

* To whom correspondence should be addressed at: CRIBI Biotechnology Centre, Università degli Studi di Padova, Via Ugo Bassi 58/B, 35121 Padova, Italy. Tel: +39 0498276221; Fax: +39 0498276280; Email: lanfra{at}cribi.unipd.it


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