Cardiac-specific overexpression of Galphaq alters excitation-contraction coupling in isolated cardiac myocytes.

:Transgenic mice with cardiac-specific overexpression of G alpha q exhibit a biochemical and physiological phenotype of load-independent cardiac hypertrophy with contractile dysfunction. To elucidate the cellular basis for altered contractility, we measured cellular contraction, Ca(2+)transients, and l -type Ca(2+)channel currents (I(Ca)) in left ventricular (LV) myocytes isolated from non transgenic (NT) controls or G alpha q hearts. Although baseline contractile function (% shortening) and the amplitude of Ca(2+)transients in G alpha q myocytes were similar to NT myocytes, the rates of cellular shortening and relengthening and the duration of Ca(2+)transients were prolonged in G alpha q myocytes. Myocytes from G alpha q hearts had larger cell capacitance but no change in I(Ca)density, voltage-dependence of activation and inactivation. The responses of I(Ca)to dihydropyridine drugs and a membrane permeable cAMP analog, 8-(4-chlorophenylthio) cAMP, were not altered; however, the time course of I(Ca)inactivation was significantly slower in G alpha q myocytes compared to NT myocytes. The kinetic difference in inactivation was abolished when Ba(2+)was used as the charge carrier or when the sarcoplasmic reticulum (SR) Ca(2+)was depleted by ryanodine, suggesting that Ca(2+)-dependent inactivation is reduced in G alpha q myocytes due to altered SR Ca(2+)release. Consistent with this hypothesis, the function of SR as assessed by the maximal Ca(2+)uptake rates and the apparent affinity of SR Ca(2+)-ATPase for Ca(2+)was reduced in ventricles of G alpha q heart. These results suggest that the reduced SR function contributes to the depressed contractility associated with this form of cardiac hypertrophy.

Yatani A ，Frank K ，Sako H ，Kranias EG ，Dorn GW 2nd ... -  《JOURNAL OF MOLECULAR AND CELLULAR CARDIOLOGY》

Enhanced Ca2+ channel currents in cardiac hypertrophy induced by activation of calcineurin-dependent pathway.

:Cardiac-specific expression of an activated calcineurin protein in the hearts of transgenic (CLN) mice produces a profound hypertrophy that rapidly progresses to heart failure. While calcineurin is regulated by Ca2+, the potential effects of calcineurin on cardiac myocyte Ca2+ handling has not been evaluated. To this end, we examined L-type Ca2+ currents (I(Ca)) in left ventricular myocytes. CLN myocytes had larger (approximately 80%) cell capacitance and enhanced I(Ca) density (approximately 20%) compared with non-transgenic (NTG) littermates, but no change in the current-voltage relationship, single-channel conductance or protein levels of alpha 1 or beta 2 subunit of L-type Ca2+ channels. Interestingly, the kinetics of I(Ca) inactivation was faster (approximately two-fold) in CLN myocytes compared with NTG myocytes. Ryanodine application slowed the rate of I(Ca) inactivation in both groups and abolished the kinetic difference, suggesting that Ca2+ dependent inactivation is increased in CLN myocytes due to altered SR Ca2+ release. Treatment of CLN mice with Cyclosporine A (CsA), a calcineurin inhibitor, prevented myocyte hypertrophy and changes in I(Ca) activity and inactivation kinetics. However, there was no direct effect of CsA on I(Ca) in either NTG or CLN myocytes, suggesting that endogenous calcineurin activity does not directly regulate Ca2+ channel activity. This interpretation is consistent with the observation that I(Ca) density, inactivation kinetics and regulation by isoproterenol were normal in cardiac-specific transgenic mice expressing calcineurin inhibitory protein domains from either Cain or AKAP79. Taken together these data suggest that chronic activation of calcineurin is associated with myocyte hypertrophy and a secondary enhancement of intracellular Ca2+ handling that is tied to the hypertrophy response itself.

Yatani A ，Honda R ，Tymitz KM ，Lalli MJ ，Molkentin JD ... -  《JOURNAL OF MOLECULAR AND CELLULAR CARDIOLOGY》

Overexpression of phospholamban alters inactivation kinetics of L-type Ca2+ channel currents in mouse atrial myocytes.

Masaki H ，Sako H ，Kadambi VJ ，Sato Y ，Kranias EG ，Yatani A ... -  《JOURNAL OF MOLECULAR AND CELLULAR CARDIOLOGY》

Calcium-handling abnormalities underlying atrial arrhythmogenesis and contractile dysfunction in dogs with congestive heart failure.

Yeh YH ，Wakili R ，Qi XY ，Chartier D ，Boknik P ，Kääb S ，Ravens U ，Coutu P ，Dobrev D ，Nattel S ... -  《-》

Reduced ryanodine receptor to dihydropyridine receptor ratio may underlie slowed contraction in a rabbit model of left ventricular cardiac hypertrophy.

Cardiac hypertrophy is associated with contractile dysfunction, a feature of which is a slowing of the time to reach peak contraction. We have examined the main mechanisms involved in the initiation of contraction and investigated if their functions are changed during cardiac hypertrophy. Cardiac hypertrophy was induced by constriction of the ascending aorta in the rabbit. After 6 weeks left ventricular myocytes were isolated or left ventricular and septal mixed membrane preparations were produced for electrophysiological and radioligand binding studies, respectively. Aortic constriction resulted in a 24% and 23% increase in heart weight to body weight ratio and cell capacitance, respectively. Action potential duration and time-to-reach 50% and 90% peak contraction (TTP(50)and TTP(90), respectively) were significantly prolonged in myocytes from hypertrophied hearts. The prolongation of TTP(50)and TTP(90)could not be explained by altered peak calcium current density or SR calcium content which were unchanged in hypertrophy. Radioligand binding studies performed on tissue preparations from the same hearts, revealed a 34% reduction in ryanodine receptor (RYR) density with no change in dihydropyridine receptor (DHPR) density. This resulted in a reduction in the ratio of RYR to DHPR from 4.4:1 to 3.3:1 in hypertrophy. Ryanodine receptor Ca(2+)-sensitivity was unchanged between sham operated and hypertrophied groups. A reduction in the ratio of RYRs to DHPRs may result in a degree of "functional uncoupling" causing defective release of Ca(2+)from the SR. These findings may underlie the slowed TTP of myocyte contraction in hypertrophy.

Milnes JT ，MacLeod KT 《JOURNAL OF MOLECULAR AND CELLULAR CARDIOLOGY》