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Cardiovascular Research Advance Access [Accepted Manuscript] published online on August 10, 2008

Cardiovascular Research, doi:10.1093/cvr/cvn225
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Published on behalf of the European Society of Cardiology. All rights reserved. © The Author 2008. For permissions please email: journals.permissions@oxfordjournals.org

Perlecan is critical for heart stability

Philipp Sasse+,1, Daniela Malan+,1,2, Michaela Fleischmann+,3, Wilhelm Roell4, Erika Gustafsson5, Toktam Bostani1,4, Yun Fan6, Thomas Kolbe3, Martin Breitbach1, Klaus Addicks7, Armin Welz4, Gottfried Brem3, Jürgen Hescheler6, Attila Aszodi8, Mercedes Costell9, Wilhelm Bloch*,2 and Bernd K. Fleischmann*,1

1 Institute of Physiology I, University of Bonn, 53105 Bonn, Germany
2 Department of Molecular and Cellular Sport Medicine, German Sport University, 50933 Cologne, Germany
3 Department of Agrobiotechnology, IFA-Tulln, Institute of Biotechnology in Animal Production, University of Natural Resources and Applied Life Sciences Vienna, 3430 Tulln, Austria
4 Department of Cardiac Surgery, University of Bonn, 53105 Bonn, Germany
5 Department of Experimental Pathology, University of Lund, 22184 Lund, Sweden
6 Institute of Neurophysiology, University of Cologne, 50931 Cologne, Germany
7 Institute of Anatomy I, University of Cologne, 50931 Cologne, Germany
8 Max Planck Institute for Biochemistry, 82152 Martinsried, Germany
9 Department of Biochemistry and Molecular Biology, University of Valencia, 46100 Burjassot, Spain

* Address correspondence to: Bernd K. Fleischmann, Institute of Physiology I, Life & Brain Center, University of Bonn, Sigmund-Freud-Strasse 25, 53105 Bonn, Tel: xx49-228-6885-200, Fax: xx49-228-6885-201, E-Mail: bernd.fleischmann{at}uni-bonn.de and Wilhelm Bloch Department of Molecular and Cellular Sport Medicine, German Sport University, Carl-Diem-Weg 6, 50933 Cologne, Tel: xx49-221-4982-5380, Fax: xx49-221-4982-8370, E-Mail: w.bloch{at}dshs-koeln.de

Aims: Perlecan is a heparansulfate proteoglycan found in basement membranes, cartilage and several mesenchymal tissues that form during development, tumor growth and tissue repair. Loss-of-function mutations in the perlecan gene in mice are associated with embryonic lethality caused primarily by cardiac abnormalities probably due to hemopericards. The aim of the present study was to investigate the mechanism underlying the early embryonic lethality and the pathophysiological relevance of perlecan for heart function.

Methods: Perlecan deficient murine embryonic stem cells were used to investigate the myofibrillar network and the electrophysiological properties of single cardiomyocytes. The mechanical stability of the developing perlecan deficient mouse hearts was analyzed by microinjecting fluorescent-labeled dextran. Maturation and formation of basement membranes and cell-cell contacts were investigated by electron microscopy, immunhistochemistry and Western Blotting.

Results: Sarcomere formation and cellular functional properties were unaffected in perlecan-deficient cardiomyocytes. However, the intraventricular dye injection experiments revealed mechanical instability of the early embryonic mouse heart muscle wall before E10.5. Accordingly, perlecan-null embryonic hearts contained lower amounts of the critical basement membrane components collagen IV and laminins. Furthermore, basement membranes were absent in perlecan-null cardiomoycytes whereas adherens junctions formed and matured around E9.5. Infarcted hearts from perlecan heterozygous mice displayed reduced heart function when compared with wild-type hearts.

Conclusions: We propose that perlecan plays an important role in maintaining the integrity during cardiac development and is important for heart function in the adult heart after injury.


Time for primary review: 27 days

+ Authors contributed equally to the ms


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