title: Analysis of the energetic state of heart cells after adenovirus-mediated expression of hALC-1.
abstract: Expression of the human atrial myosin light chain 1 (hALC-1) in the cardiac ventricle in vivo as well as in primary cultivated adult cardiomyocytes caused a pronounced positive inotropic effect. Therefore, it is one of the most promising candidate gene to treat congestive heart failure (CHF). In this work, we investigated, whether hALC-1 expression also modifies the energetic state of cardiomyocytes. Primary cultivated neonatal rat hearts cells (NRHC) were infected with adenoviral vectors (Ad vectors) containing a hALC-1 cDNA (AdCMV.hALC-1) or a control Ad vector. Infection efficiency of NRHC reached 100% at 50 multiplicity of infection (MOI). Interestingly and in contrast to primary cultures of liver cells, there were no cytotoxic side effects or induction of apoptosis up to MOI 50 in Ad vector infected NRHC. NRHC expressed large amounts of hALC-1 upon infection with AdCMV.hALC-1 which could easily been detected by protein staining and Western blot analysis. Analysis of intracellular hALC-1 localization by double-labeling immunofluorescence of AdCMV.hALC-1 infected cardiomyocytes revealed the typical myofibrillar striation pattern, as well as co-localization of hALC-1 with myosin heavy chains. There was no difference in the oxygen consumption between controls and AdCMV.hALC-1 infected NRHC. These data suggest that first: adenoviral vectors could be used as a safe and effective tool for gene transfer to cardiomyocytes, and second: that a positive inotropic effect of hALC-1 is not associated with enhanced oxygen consumption.
authors.affiliation: Max Delbrück Center for Molecular Medicine (MDC), 13122 Berlin, Germany.
authors.forename: Udo B
authors.initials: UB
authors.lastname: Zacharzowsky
authors.forename: Gerhard
authors.initials: G
authors.lastname: Wolff
authors.forename: Monika
authors.initials: M
authors.lastname: Kott
authors.forename: Hannelore
authors.initials: H
authors.lastname: Haase
authors.forename: Holger
authors.initials: H
authors.lastname: Bartsch
authors.forename: Andreas K
authors.initials: AK
authors.lastname: Nuessler
authors.forename: Leonidas G
authors.initials: LG
authors.lastname: Baltas
authors.forename: Leonid
authors.initials: L
authors.lastname: Karawajew
authors.forename: Ingo
authors.initials: I
authors.lastname: Morano
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D017209:Apoptosis; D024743:Atrial Myosins / Q000235:genetics / Q000378:metabolism*; D002453:Cell Cycle; D002478:Cells, Cultured; D003588:Cytopathogenic Effect, Viral; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D006321:Heart / Q000502:physiology* / Q000821:virology; D006801:Humans; D009206:Myocardium / Q000166:cytology* / Q000378:metabolism*; D018994:Myosin Light Chains / Q000235:genetics / Q000378:metabolism*; D010101:Oxygen Consumption; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D018994:Myosin Light Chains; C491915:hALC-1 protein, human; D024743:Atrial Myosins