title: Somatic mosaicism, germline expansions, germline reversions and intergenerational reductions in myotonic dystrophy males: small pool PCR analyses.
abstract: In order to characterize the dynamics of CTG repeat instability in somatic and germline tissue from myotonic dystrophy (DM) males we have used small pool polymerase chain reaction (PCR) in a detailed quantitative analysis of repeat length variation. We demonstrate that the heterogeneous smear of CTG repeats observed in DM patients using standard analyses is comprised of multiple unresolved bands that may be dissected into discrete length alleles derived from single cells using single molecule PCR techniques. Analysis of somatic tissues demonstrates a bias toward increasing allele length and a lower boundary below which variant alleles are rare, consistent with a highly directional expansion pathway in the soma. Two sperm samples show extensive variation and a size increase bias, concordant with the phenomenon of anticipation. In addition, sperm analysis shows that large contractions, including reversions into the normal size range, are restricted to the germline. Detailed analysis of intergenerational 'reductions' paternally transmitted to two offspring suggests that some apparent reductions may be artifacts of somatic expansion in the parent. Our data indicate that in addition to germline variation, substantial somatic expansion can also contribute to the intergenerational differences usually observed in DM.
authors.affiliation: Department of Molecular and Human Genetics, Baylor College of Medicine, Texas Medical Center, Houston, TX 77030, USA.
authors.forename: D G
authors.initials: DG
authors.lastname: Monckton
authors.forename: L J
authors.initials: LJ
authors.lastname: Wong
authors.forename: T
authors.initials: T
authors.lastname: Ashizawa
authors.forename: C T
authors.initials: CT
authors.lastname: Caskey
mesh_terms: D000328:Adult; D001483:Base Sequence; D002648:Child; D017931:DNA Primers; D018095:Germ-Line Mutation; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D009030:Mosaicism; D009132:Muscles / Q000378:metabolism; D009223:Myotonic Dystrophy / Q000097:blood / Q000235:genetics*; D016133:Polymerase Chain Reaction; D013094:Spermatozoa / Q000378:metabolism
chemical_list: D017931:DNA Primers