title: Cocaine activates calcium/calmodulin kinase II and causes cardiomyocyte hypertrophy.
abstract: Cardiac hypertrophy occurs in as many as 47% of normotensive individuals who chronically use cocaine. We investigated the effects of cocaine, in concentrations commonly found in chronic cocaine users, on calcium/calmodulin kinase (CaMK), and whether cocaine can activate CaMK, increase cardiac myocyte protein expression, and cause cardiac hypertrophy in this manner. In series I to III, 0 (control) or cocaine in concentrations of 10 to 10 mol/L was added to cultured adult rat cardiac ventricular myocytes to determine by Western blots and by P incorporation the optimal treatment time and the optimal dose for CaMK activation. In series I, cocaine, 10 mol/L, increased myocyte CaMKII translocation from myocyte soluble to particulate fractions by > or =73 +/- 9% (P < 0.01) in comparison with controls but did not cause the translocation of CaMKI or CaMKIV. In series II and III, cocaine treatment of myocytes for 15 minutes increased maximal CaMKII activity by 86.5 +/- 13.3% (P < 0.001) and a cocaine dose of 5 x 10 mol/L increased CaMKII activity by 169.5 +/- 18.1% (P < 0.001). In series IV we measured by silver staining beta-myosin heavy chain protein (beta-MHC) expression in myocytes before and after cocaine and also CaMK inhibition with KN-62 (1-[N,O-bis-(5-isoquinolinesulfonyl)-N-methyl-L-tyrosyl]-4-phenylpiperazine). In these experiments, cocaine, 5x10 mol/L, increased myocyte protein concentration by 29.2 +/- 2.8%, and beta-MHC by 93.2 +/- 8.8% (P < 0.001). In series V and VI, cocaine effects on calcium currents (ICa) and intracellular Ca ([Ca]i) were determined before and after CaMK inhibition with KN-62 in rat myocytes. Cocaine, 10 mol/L, enhanced ICa peak amplitude in a voltage-dependent manner (by 173.9 +/- 14.9% at -20 mV and by 38.4 +/- 6.9% at 0 mV P < 0.01). Cocaine, 10 to 10 mol/L, in series VI promoted Ca transients from myocyte sarcoplasmic reticulum and increased [Ca]i to 607 +/- 141 x 10 mol/L (P < 0.05). KN-62 decreased cocaine-induced myocyte protein expression by 76.6%, and beta-MHC by 66.2% (P < 0.01) and significantly decreased cocaine-induced Ca transients and [Ca]i. We conclude that CaMKII activation is an important mechanism whereby cocaine can cause myocyte hypertrophy.
authors.affiliation: Department of Medicine, University of South Florida College of Medicine and the James A. Haley VA Hospital, Tampa, Florida 33612, USA. rhenning@hsc.usf.edu
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Henning
authors.forename: Javier
authors.initials: J
authors.lastname: Cuevas
mesh_terms: D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000031:analogs & derivatives / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D015220:Calcium Channels / Q000502:physiology; D020013:Calcium Signaling / Q000187:drug effects; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D002478:Cells, Cultured; D003042:Cocaine / Q000633:toxicity*; D004305:Dose-Response Relationship, Drug; D006352:Heart Ventricles / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D006984:Hypertrophy; D008297:Male; D008564:Membrane Potentials / Q000187:drug effects; D032383:Myocytes, Cardiac / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology; D018995:Myosin Heavy Chains / Q000378:metabolism; D018408:Patch-Clamp Techniques; D010455:Peptides / Q000378:metabolism; D010656:Phenylephrine / Q000494:pharmacology; D010761:Phosphorus Radioisotopes; D010766:Phosphorylation / Q000187:drug effects; D051381:Rats; D017207:Rats, Sprague-Dawley; D012508:Sarcolemma / Q000187:drug effects / Q000502:physiology
chemical_list: D015220:Calcium Channels; D010455:Peptides; D010761:Phosphorus Radioisotopes; C085903:autocamtide-2; D010656:Phenylephrine; C063302:KN 62; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D000255:Adenosine Triphosphate; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D018995:Myosin Heavy Chains; D003042:Cocaine