Abstract
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 (F(init)) (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, F(init) (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 β-galactosidase and Luc coding for firefly luciferase. Luciferase activity increased more than 2,500-fold from background levels of 8.7 x 103 ± 5.0 x 103 relative light units (RLU)/mg protein (from hearts transfected with promotorless adenovirus with luciferase transgene construct AdNULLLuc, n = 5) to 23.4 x 106 ± 11.1 x 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.
| Original language | English (US) |
|---|---|
| Journal | American Journal of Physiology - Heart and Circulatory Physiology |
| Volume | 279 |
| Issue number | 3 48-3 |
| State | Published - 2000 |
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Keywords
- Adenovirus
- Rabbit
- Trabecula
ASJC Scopus subject areas
- Physiology
- Physiology (medical)
Cite this
Preservation of myocardial function after adenoviral gene transfer in isolated myocardium. / Lehnart, S. E.; Janssen, P. M L; Franz, W. M.; Donahue, J. K.; Lawrence, J. H.; Marbán, E.; Prestle, J.; Hasenfuss, G.
In: American Journal of Physiology - Heart and Circulatory Physiology, Vol. 279, No. 3 48-3, 2000.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Preservation of myocardial function after adenoviral gene transfer in isolated myocardium
AU - Lehnart, S. E.
AU - Janssen, P. M L
AU - Franz, W. M.
AU - Donahue, J. K.
AU - Lawrence, J. H.
AU - Marbán, E.
AU - Prestle, J.
AU - Hasenfuss, G.
PY - 2000
Y1 - 2000
N2 - 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 (F(init)) (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, F(init) (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 β-galactosidase and Luc coding for firefly luciferase. Luciferase activity increased more than 2,500-fold from background levels of 8.7 x 103 ± 5.0 x 103 relative light units (RLU)/mg protein (from hearts transfected with promotorless adenovirus with luciferase transgene construct AdNULLLuc, n = 5) to 23.4 x 106 ± 11.1 x 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.
AB - 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 (F(init)) (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, F(init) (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 β-galactosidase and Luc coding for firefly luciferase. Luciferase activity increased more than 2,500-fold from background levels of 8.7 x 103 ± 5.0 x 103 relative light units (RLU)/mg protein (from hearts transfected with promotorless adenovirus with luciferase transgene construct AdNULLLuc, n = 5) to 23.4 x 106 ± 11.1 x 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.
KW - Adenovirus
KW - Rabbit
KW - Trabecula
UR - http://www.scopus.com/inward/record.url?scp=0033826425&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0033826425&partnerID=8YFLogxK
M3 - Article
C2 - 10993759
AN - SCOPUS:0033826425
VL - 279
JO - American Journal of Physiology
JF - American Journal of Physiology
SN - 0363-6135
IS - 3 48-3
ER -