Severe Myocardial Fibrosis Caused by a Deletion of the 5' End of the Lamin A/C Gene
J. Peter van Tintelen, MD*,*,
Rene A. Tio, MD, PhD ,
Wilhelmina S. Kerstjens-Frederikse, MD*,
Jop H. van Berlo, MD ,
Ludolf G. Boven, BSc*,
Albert J.H. Suurmeijer, MD, PhD ,
Stefan J. White, PhD||,
Johan T. den Dunnen, PhD||,
Gerard J. te Meerman, PhD*,
Yvonne J. Vos, MSc*,
Annemarie H. van der Hout, PhD*,
Jan Osinga*,
Maarten P. van den Berg, MD, PhD,
Dirk J. van Veldhuisen, MD, PhD,
Charles H.C.M. Buys, PhD*,
Robert M.W. Hofstra, PhD* and
Yigal M. Pinto, MD, PhD
* Department of Genetics, University Medical Center Groningen, University of Groningen, Groningen, the Netherlands
Department of Cardiology, University Medical Center Groningen, University of Groningen, Groningen, the Netherlands
Department of Pathology, University Medical Center Groningen, University of Groningen, Groningen, the Netherlands
Department of Cardiology, University Hospital Maastricht, Maastricht, the Netherlands
|| Department of Human and Clinical Genetics, Leiden University Medical Center, Leiden, the Netherlands
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Figure 1 Pedigree of the Family
Solid symbols indicate clinically and genetically affected persons (deletion carriers). One-quarter black-filled symbols indicate those clinically affected upon personal examination or from medical records or literature (11). Open symbols indicate clinically unaffected individuals and line-through symbols indicate deceased individuals. Squares represent men, and circles represent women. *The respective person has been evaluated genetically. The haplotypes are represented in Table 2. SCD = sudden cardiac death.
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Figure 3 Southern Blot Analysis of LMNA Deletion Carriers and Control Subjects
Southern blot showing reduced intensities of the signal corresponding with the start codon–containing exon of the lamin AC (LMNA) gene in affected patients (IV-8, IV-9, IV-11) as compared with control subjects (C) and a nonaffected family member (IV-10).
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Figure 4 PCR on Complementary Deoxyribonucleic Acid
Reverse transcription polymerase chain reaction (RT-PCR) on ribonucleic acid (RNA) extracted from fibroblasts of patient III-12 and from lymphocytes of a control individual. The RNA of the patient show, in addition to the normal-sized band, a shorter product that after sequencing proved to have lost both exons –1 and +1. The normal-sized product is weaker, owing to the fact that shorter amplicons are more abundantly amplified.
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Figure 5 Sequence Analysis of cDNA in Deletion Carrier and Control Subjects
Forward complementary deoxyribonucleic acid (cDNA) sequence and schematic representation of the 5' region of the lamin AC gene on cDNA level in the wild type (A) and mutated situation (B; patient III-12). mRNA = messenger ribonucleic acid.
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Figure 6 Western Blot Analysis of Lamins A and C in Fibroblasts
Western blot analysis showing decreased expression of both lamins A and C in fibroblasts from mutation carrier (B), as compared with a non-carrier family member (A). No alternative product from the mutated allele was seen. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) serves as a control that equal amounts of protein have been loaded.
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Figure 7 Indirect Immunofluorescence Assay With Lamin A Antibody
Indirect immunofluorescence with lamin A antibody showing nuclear lamin aggregates in fibroblasts from mutation carrier but not in control fibroblasts. DAPI = 4'-6-diamidino-2-phenylindole.
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