Endothelin-1 Responsiveness of a 1.4 kb Phospholamban Promoter Fragment in Rat Cardiomyocytes Transfected by the Gene Gun

• Published in: Journal of molecular and cellular cardiology 2000-02, Vol.32 (2), p.311-321
• Main Authors: Eizema, Karin, van Heugten, Han A.A., Bezstarosti, Karel, van Setten, Marga C., Lamers, Jos M.J.
• Format: Article
• Language: English
• Subjects: Animals; Animals, Newborn; Atrial Natriuretic Factor - biosynthesis; Atrial Natriuretic Factor - genetics; Atrial natriuretic peptide; Base Sequence; Biolistics; Biolistics - instrumentation; Ca2+-pump; Calcium-Binding Proteins - biosynthesis; Calcium-Binding Proteins - genetics; Cells, Cultured - drug effects; Culture Media, Serum-Free; Endothelin-1; Endothelin-1 - pharmacology; Gene Expression Regulation - drug effects; Genes, Reporter; Heart - drug effects; Helios Gene Gun; Hypertrophy; Luciferases - biosynthesis; Luciferases - genetics; Molecular Sequence Data; Muscle Proteins - biosynthesis; Muscle Proteins - genetics; Myocardium - cytology; Phospholamban; Promoter Regions, Genetic; Rats; Recombinant Fusion Proteins - biosynthesis; Transcription, Genetic; Transfection - instrumentation; Transfection - methods
• ISSN: 0022-2828; 1095-8584
• DOI: 10.1006/jmcc.1999.1076

The transcriptional regulation of an isolated rat phospholamban (PL) promoter fragment in rat cardiomyocytes was analyzed by applying a new method to reach substantially higher transfection efficiencies: gene gun biolistics. The gene gun transfection method was optimized for application to primary cultures of rat neonatal cardiomyocytes. Cells, cultured at different densities (0.75–1.50×105cells/cm2) in serum-free medium, were transfected with DNA coated gold particles. A transfection efficiency of up to 10% could be achieved (compared to