Document Detail

Robust adenoviral and adeno-associated viral gene transfer to the in vivo murine heart: application to study of phospholamban physiology.
MedLine Citation:
PMID:  14638552     Owner:  NLM     Status:  MEDLINE    
BACKGROUND: Viral gene transfer to the whole heart in vivo has been achieved in several mammalian species but remained difficult to accomplish in murine hearts. We postulated that a key impediment derives from the use of proximal aortic occlusion during virus injection, because this eliminates coronary perfusion gradients in mice as aortic root and left ventricle pressures equalize. METHODS AND RESULTS: Pressure-volume analysis confirmed these mechanics. In contrast, descending aortic occlusion with whole-body cooling (20 degrees C) preserved transmyocardial perfusion gradients and allowed for sustained (>10-minute) dwell times in an upper-body perfusion circuit. This approach yielded robust cardiac transfection with adenovirus (AdV) and adeno-associated virus (AAV) injected into the left ventricle cavity or more simply via a central vein. Cardio-specific expression was achieved with a myocyte-specific promotor. Optimal AdV transfection required 9-minute aortic occlusion, versus 5-minute occlusion for AAV. Using this method, we examined the in vivo function of phospholamban (PLB) by stably transfecting PLB-null mice with AAV encoding PLB (AAV(PLB)). AAV(PLB) restored PLB protein to near control levels that colocalized with SERCA2A in cardiomyocytes. At baseline, PLB-null hearts exhibited enhanced systolic and diastolic function, but frequency-dependent reserve was blunted versus wild-type controls. These properties, particularly the frequency response, returned toward control 3 months after AAV(PLB) transfection. CONCLUSIONS: The new simplified approach for murine whole-heart viral transfection should assist molecular physiology studies.
Hunter C Champion; Dimitrios Georgakopoulos; Saptarsi Haldar; Lili Wang; Yibin Wang; David A Kass
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.     Date:  2003-11-24
Journal Detail:
Title:  Circulation     Volume:  108     ISSN:  1524-4539     ISO Abbreviation:  Circulation     Publication Date:  2003 Dec 
Date Detail:
Created Date:  2003-12-05     Completed Date:  2004-01-09     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0147763     Medline TA:  Circulation     Country:  United States    
Other Details:
Languages:  eng     Pagination:  2790-7     Citation Subset:  AIM; IM    
Division of Cardiology, Department of Medicine, 600 N Wolfe St, Halsted 500, Johns Hopkins Hospital, Baltimore, Md 21287, USA.
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MeSH Terms
Adenoviridae / genetics*
Aorta / physiology
Calcium-Binding Proteins / deficiency,  genetics,  physiology*
Calcium-Transporting ATPases / biosynthesis
Cell Separation
Coronary Circulation
Dependovirus / genetics*
Gene Transfer Techniques*
Heart / physiology*,  virology
Mice, Knockout
Myocytes, Cardiac / metabolism
Sarcoplasmic Reticulum Calcium-Transporting ATPases
Time Factors
Ventricular Function, Left / genetics,  physiology
beta-Galactosidase / biosynthesis,  genetics
Grant Support
Reg. No./Substance:
0/Calcium-Binding Proteins; 0/phospholamban; EC; EC ATPases; EC Reticulum Calcium-Transporting ATPases

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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