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1 Abteilung Kardiologie und Pneumologie, Universität Göttingen, D-37075 Göttingen, Germany; 2 Medizinische Klinik II, Universität zu Lübeck, D-23538 Lübeck, Germany; and 3 Section of Molecular and Cellular Cardiology, Johns Hopkins University Medical School, Baltimore, Maryland 21205
Adenoviral gene transfer to the heart
represents a promising model for structure-function analyses. Rabbit
hearts were subjected to an ex vivo perfusion protocol that achieves
gene transfer in >90% of cardiac myocytes. Contractile function of
isolated myocardial preparations of these hearts was then observed for
2 days in a recently developed trabecula culture system. In
sham-infected hearts, the initial developed force
(Finit) (15.6 ± 3.7 mN/mm2;
n = 12) did not change significantly after 48 h
(17.0 ± 1.9 mN/mm2; P = 0.46). In
adenovirus-infected preparations, Finit (14.3 ± 1.8 mN/mm2; n = 21) did not significantly
differ from the control (P = 0.75) and was unchanged
after 48 h (15.3 ± 2.5 mN/mm2; P = 0.93). After 2 days of continuous contractions, we observed homogenous and high-level expression of the reporter genes
LacZ coding for 
-galactosidase and Luc coding
for firefly luciferase. Luciferase activity increased more than
2,500-fold from background levels of 8.7 × 103 ±
5.0 × 103 relative light units (RLU)/mg protein (from
hearts transfected with promotorless adenovirus with luciferase
transgene construct AdNULLLuc, n = 5) to
23.4 × 106 ± 11.1 × 106 RLU/mg
protein (from hearts tranfected with adenovirus with Rous sarcoma virus
promotor and luciferase transgene construct AdRSVLuc, n = 5) in infected myocardial preparations
(P < 0.005). Our results demonstrate a new ex vivo
approach to achieve homogenous and high-level expression of recombinant
adenoviral genes in contracting myocardium without adverse functional effects.
adenovirus; trabecula; rabbit
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