Ribonuclear inclusions as biomarker of myotonic dystrophy type 2, even in improperly frozen or defrozen skeletal muscle biopsies.
Myotonic dystrophy type 2 (DM2) is a dominantly inherited disorder caused by a CCTG repeat expansion in intron 1 of ZNF9 gene. The size and the somatic instability of DM2 expansion complicate the molecular diagnosis of DM2. In situ hybridization represents a rapid and sensitive method to obtain a definitive diagnosis in few hours, since it allows the direct visualization of the mutant mRNA foci on skeletal muscle sections. This approach makes the muscle biopsy an important tool for definitive diagnosis of DM2. Consequently, a rapid freezing at ultra cold temperature and a good storage of muscle specimens are essential to avoid morphologic alterations and nucleic acids degradation. However incorrect freezing or thawing may accidentally occur. In this work we report that fluorescence in situ hybridization may be applied on improperly frozen or inappropriately stored muscle biopsies since foci of mutant mRNA are well preserved and can still be detected in muscle sections no more useful for histopathological evaluation.
Cardani
R
R
Department of Molecular Biology and Biotechnologies, University of Milan, Italy.
Mancinelli
E
E
Giagnacovo
M
M
Sansone
V
V
Meola
G
G
D015415:Biomarkers / Q000032:analysis; D001706:Biopsy; D002467:Cell Nucleus / Q000378:metabolism / Q000473:pathology; D015925:Cryopreservation; D042622:DNA Repeat Expansion / Q000235:genetics*; D005615:Freezing; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D018656:Muscle Fibers, Fast-Twitch / Q000378:metabolism / Q000473:pathology; D018657:Muscle Fibers, Slow-Twitch / Q000378:metabolism / Q000473:pathology; D018482:Muscle, Skeletal / Q000378:metabolism* / Q000473:pathology; D009223:Myotonic Dystrophy / Q000175:diagnosis* / Q000235:genetics* / Q000473:pathology; D012333:RNA, Messenger / Q000235:genetics; D016601:RNA-Binding Proteins / Q000235:genetics; D013048:Specimen Handling / Q000379:methods*
D015415:Biomarkers; C433227:CNBP protein, human; D012333:RNA, Messenger; D016601:RNA-Binding Proteins
Neuromuscul Disord. 2008 Aug;18(8):641-5
18653337
Neuromuscul Disord. 2003 Sep;13(7-8):589-96
12921797
Am J Hum Genet. 2003 Oct;73(4):835-48
12970845
Eur J Histochem. 2005 Jan-Mar;49(1):93-6
15823800
Biochim Biophys Acta. 2006 Mar;1762(3):329-34
16376058
Hum Mol Genet. 2001 Sep 15;10(19):2165-70
11590133
Eur J Histochem. 2004 Oct-Dec;48(4):437-42
15718211
J Biol Chem. 2005 Feb 18;280(7):5773-80
15546872
Science. 2001 Aug 3;293(5531):864-7
11486088
Mol Cell. 2002 Jul;10(1):45-53
12150906
Neuromuscul Disord. 2004 Apr;14(4):274-83
15019706
Hum Mol Genet. 2006 Jun 1;15(11):1808-15
16624843
Am J Hum Genet. 2004 Jun;74(6):1309-13
15114529
Mol Cell. 2002 Jul;10(1):35-44
12150905
Hum Mol Genet. 2002 Apr 1;11(7):805-14
11929853
J Neuropathol Exp Neurol. 2008 Apr;67(4):319-25
18379436
Hum Mol Genet. 2006 Oct 15;15 Spec No 2:R162-9
16987879
Science. 2004 Jan 16;303(5656):383-7
14657503
Neuromuscul Disord. 2005 Jan;15(1):5-16
15639115
Neurology. 2003 Jun 10;60(11):1854-7
12796551