Chapter 4. Materials and methods

Table of Contents
4.1. Materials
4.2. Experimental animals
4.3. Isolated perfused heart preparations (I-V)
4.4. Exercise experiment (V)
4.5. Experimental protocols
4.6. Isolation and analysis of cytoplasmic RNA (I-III, V)
4.7. Radioimmunoassays (I- III)
4.8. Cyclic AMP measurements (V)
4.9. Analysis of markers for cardiac injury (V)
4.10. Histology (IV, V)
4.11. Statistical analysis

4.1. Materials

The chemicals and supplies used in this study were: formaldehyde and guanidine isothiocyanate (Fluka Chemie AG, Buchs, Switzerland), CsCl (Serva Feinchemica Gmbh & Co, Heidelberg, Germany), LiCl (JT Baker Chemicals BV, Holland), agarose NA (Pharmacia LKB Biotechnology, Uppsala, Sweden), Hybond, N+ nylon membrane (Amersham Life Science, Buckinghamshire, UK), [32P]-deoxycytidine-5´-triphosphate (dCTP, Amersham), Quick Prime Kit (Pharmacia, Sweden), X-ray films (Eastman Kodak, Rochester, NY, USA, and Amersham), ET-1 (Phoenix Pharmaceuticals Inc., CA, USA and Peninsula Laboratories, Belmont, CA, USA), rat AM-(1-50) (Phoenix Pharmaceuticals), isoproterenol, saponin and Nω -nitro-L-arginine methyl ester (L-NAME, Sigma Chemical Company, St. Louis, Missouri, USA.), BQ-123 and BQ-788 (Phoenix Pharmaceuticals). Bosentan was generously supplied by Dr. Martine Clozel (F. Hoffmann-La Roche Ltd., Basel, Switzerland and Actelion Ltd., Allschwil, Switzerland) and CV-11974 by Dr. Hajime Toguchi (Takeda Chemical Industries Ltd, Osaka, Japan). Other chemicals were from Sigma.