Molecular characterization of the stretch-induced adaptation of cultured cardiac cells. An in vitro model of load-induced cardiac hypertrophy.
Although it is a well-known fact that hemodynamic load is a major determinant of cardiac muscle mass and its phenotype, little is known as to how mechanical load is converted into intracellular signals of gene regulation. To address this question, we characterized the stretch-induced adaptation of cultured neonatal cardiocytes grown on a stretchable substrate in a serum-free medium. Static stretch (20%) of the cells was applied without cell injury. Stretch caused hypertrophy in myocytes and hyperplasia in non-myocytes. Stretch caused an induction of immediate-early genes such as c-fos, c-jun, c-myc, JE, and Egr-1, but not Hsp70. Immunostaining showed that the stretch-induced Fos protein localized in the nucleus of both myocytes and non-myocytes. Nuclear extracts from stretched myocytes contained DNA binding activity to the AP-1 and Egr-1 consensus sequences. In myocytes, the induction of immediate-early genes was followed by expression of "fetal" genes such as skeletal alpha-actin, atrial natriuretic factor, and beta-myosin heavy chain. DNA transfection experiments showed that the "stretch-response element" of the c-fos gene promoter is present within 356 base pairs of the 5'-flanking region, whereas that of the atrial natriuretic factor and the beta-myosin heavy chain genes is probably located outside of 3412 and 628 base pairs of the 5'-flanking region, respectively. These results demonstrate that the phenotype of stretched cardiocytes in this in vitro model closely mimics that of hemodynamic load-induced hypertrophy in vivo. This model seems to be a suitable system with which to dissect the molecular mechanisms of load-induced hypertrophy of cardiac muscle.
Sadoshima
J
J
Molecular Medicine Unit, Beth Israel Hospital, Boston, Massachusetts 02215.
Jahn
L
L
Takahashi
T
T
Kulik
T J
TJ
Izumo
S
S
D000222:Adaptation, Physiological; D001769:Blood; D015152:Blotting, Northern; D006332:Cardiomegaly / Q000473:pathology / Q000503:physiopathology*; D002478:Cells, Cultured; D003402:Creatine Kinase / Q000378:metabolism; D016895:Culture Media, Serum-Free; D005455:Fluorescent Antibody Technique; D016762:Genes, fos; D016758:Genes, jun; D016259:Genes, myc; D006321:Heart / Q000503:physiopathology*; D006360:Heat-Shock Proteins / Q000235:genetics; D006439:Hemodynamics; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D009206:Myocardium / Q000201:enzymology / Q000473:pathology*; D009218:Myosins / Q000235:genetics; D010649:Phenylalanine / Q000378:metabolism; D010957:Plasmids; D013936:Thymidine / Q000378:metabolism
D016895:Culture Media, Serum-Free; D006360:Heat-Shock Proteins; D010649:Phenylalanine; D007770:L-Lactate Dehydrogenase; D003402:Creatine Kinase; D009218:Myosins; D013936:Thymidine