Mechanical stress stimulates phospholipase C activity and intracellular calcium ion levels in neonatal rat cardiomyocytes.
To investigate how mechanical stress is sensed by cardiomyocytes and translated to cardiac hypertrophy, cardiomyocytes were subjected to stretch while measuring phospholipase C (PLC) and phospholipase D (PLD) activities and levels of intracellular calcium ions ([Ca2+]i) and pH. In stretched cardiomyocytes, PLC activity increased 2-fold after 30 min, whereas PLD activity hardly increased at all. Mechanical stress induced by prodding or by cell stretch increased [Ca2+](i)by a factor 5.2 and 4, respectively. Gadolinium chloride (stretch-activated channel blocker) attenuated the prodding-induced and stretch-induced [Ca2+](i)rise by about 50%. Blockade of ryanodine receptors by a combination of Ruthenium Red and procaine reduced the [Ca2+](i)rise only partially. Diltiazem (L-type Ca2+ channel antagonist) blocked the prodding-induced [Ca2+](i)rise completely, and reduced the stretch-induced [Ca2+](i)rise by about 50%. The stretch-induced [Ca2+](i)rise was unaffected by U73122, an inhibitor of PLC activity. Stretch did not cause cellular alkalinization. In conclusion, in cardiomyocytes, PLC and [Ca2+](i)levels are involved in the stretch-induced signal transduction, whereas PLD plays apparently no role. The stretch-induced rise in [Ca2+](i)in cardiomyocytes is most probably caused by [Ca2+](i)influx through L-type Ca2+ channels and stretch-activated channels, leading to Ca2+-induced Ca2+ -release from the SR via the ryanodine receptor.
Ruwhof
C
C
Department of Cardiology, Leiden University Medical Center, Leiden, The Netherlands.
van Wamel
J T
JT
Noordzij
L A
LA
Aydin
S
S
Harper
J C
JC
van der Laarse
A
A
D000779:Anesthetics, Local / Q000494:pharmacology; D000818:Animals; D000831:Animals, Newborn; D000893:Anti-Inflammatory Agents / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D002121:Calcium Channel Blockers / Q000494:pharmacology; D006332:Cardiomegaly / Q000209:etiology; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D004110:Diltiazem / Q000494:pharmacology; D005456:Fluorescent Dyes / Q000378:metabolism; D005682:Gadolinium / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents / Q000494:pharmacology; D009206:Myocardium / Q000166:cytology* / Q000378:metabolism; D010739:Phospholipase D / Q000378:metabolism; D011343:Procaine / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D012430:Ruthenium Red / Q000494:pharmacology; D013314:Stress, Mechanical; D010738:Type C Phospholipases / Q000037:antagonists & inhibitors / Q000378:metabolism*
D000779:Anesthetics, Local; D000893:Anti-Inflammatory Agents; D002121:Calcium Channel Blockers; D016895:Culture Media, Serum-Free; D005456:Fluorescent Dyes; D007202:Indicators and Reagents; D012430:Ruthenium Red; D011343:Procaine; D005682:Gadolinium; D010738:Type C Phospholipases; D010739:Phospholipase D; D004110:Diltiazem; C038958:gadolinium chloride; D002118:Calcium