Distribution and function of recombinant endothelial nitric oxide synthase in transplanted hearts

J. Yap, T. O'Brien, C. Pellegrini, D. A. Barber, H. D. Tazelaar, S. R. Severson, Virginia M Miller, C. G A McGregor

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Introducing recombinant genes into donor hearts may offer a therapeutic intervention that could potentially attenuate the complications of heart transplantation, including rejection, infection and accelerated atherosclerosis. In the cardiovascular system, reduced bioactivity of endothelial nitric oxide is a feature of atherosclerosis and vascular injury. Nitric oxide is an arterial vasodilator that also inhibits proliferation of vascular smooth muscle cells and platelet aggregation. Experiments were designed to determine the distribution of adenoviral-mediated transfer of recombinant endothelial nitric oxide synthase gene (eNOS) and the effect of recombinant gene expression on the function of transplanted hearts. Adenoviral vectors for (a) bovine eNOS (AdeNOS) or (b) β-galactosidase (AdLacZ; control) were infused into two groups (n = 12, per group) of explanted rat hearts. The transduced hearts were then implanted heterotopically into the abdomen of syngeneic recipient rats. After four days, the hearts were excised and examined for distribution and function of the recombinant genes. Polymerase chain reaction (pCR) verified the presence of the recombinant eNOS gene in eNOS-transduced but not in β-galactosidase- transduced hearts; reverse transcriptase-PCR identified mRNA for eNOS in AdeNOS-transduced hearts. NOS activity (conversion of tritiated L-arginine to citrulline) was greater in homogenates of AdeNOS- compared to AdLacZ- transduced hearts. Positive immunoreactivity for eNOS was present in cardiomyocytes predominantly in eNOS-transduced hearts. Myocardial contractility and coronary blood flow, as determined using a Langendorff preparation, were not different between hearts transduced with AdeNOS or AdLacZ. These results suggest that, up to four days post transplantation, adenoviral-mediated transfer of eNOS into transplanted hearts is possible. However, expression of the recombinant protein did not result in measurable changes in myocardial contractility or coronary perfusion.

Original languageEnglish (US)
Pages (from-to)720-727
Number of pages8
JournalCardiovascular Research
Volume42
Issue number3
DOIs
StatePublished - Jun 1999

Fingerprint

Nitric Oxide Synthase Type III
Genes
Galactosidases
Atherosclerosis
Nitric Oxide
Citrulline
Cell Aggregation
Vascular System Injuries
Graft Rejection
Heart Transplantation
Cardiovascular System
Reverse Transcriptase Polymerase Chain Reaction
Vasodilator Agents
Platelet Aggregation
Vascular Smooth Muscle
Recombinant Proteins
Cardiac Myocytes
Abdomen
Smooth Muscle Myocytes
Arginine

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine

Cite this

Yap, J., O'Brien, T., Pellegrini, C., Barber, D. A., Tazelaar, H. D., Severson, S. R., ... McGregor, C. G. A. (1999). Distribution and function of recombinant endothelial nitric oxide synthase in transplanted hearts. Cardiovascular Research, 42(3), 720-727. https://doi.org/10.1016/S0008-6363(99)00010-3

Distribution and function of recombinant endothelial nitric oxide synthase in transplanted hearts. / Yap, J.; O'Brien, T.; Pellegrini, C.; Barber, D. A.; Tazelaar, H. D.; Severson, S. R.; Miller, Virginia M; McGregor, C. G A.

In: Cardiovascular Research, Vol. 42, No. 3, 06.1999, p. 720-727.

Research output: Contribution to journalArticle

Yap, J, O'Brien, T, Pellegrini, C, Barber, DA, Tazelaar, HD, Severson, SR, Miller, VM & McGregor, CGA 1999, 'Distribution and function of recombinant endothelial nitric oxide synthase in transplanted hearts', Cardiovascular Research, vol. 42, no. 3, pp. 720-727. https://doi.org/10.1016/S0008-6363(99)00010-3
Yap, J. ; O'Brien, T. ; Pellegrini, C. ; Barber, D. A. ; Tazelaar, H. D. ; Severson, S. R. ; Miller, Virginia M ; McGregor, C. G A. / Distribution and function of recombinant endothelial nitric oxide synthase in transplanted hearts. In: Cardiovascular Research. 1999 ; Vol. 42, No. 3. pp. 720-727.
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