Primary culture of human atrial myocytes is associated with the appearance of structural and functional characteristics of immature myocardium

被引:39
作者
Benardeau, A
Hatem, SN
RuckerMartin, C
Tessier, S
Dinanian, S
Samuel, JL
Coraboeuf, E
Mercadier, JJ
机构
[1] UNIV PARIS 11, CNRS URA 1159, HOP MARIE LANNELONGUE, LAB CARDIOL MOL & CELLULAIRE, F-92350 LE PLESSIS ROBINSON, FRANCE
[2] HOP LARIBOISIERE, INSERM U127, F-75475 PARIS, FRANCE
关键词
primary culture; human atrial myocytes; ionic currents; action potential; myocyte de-differentiation; myocyte immunocytochemistry;
D O I
10.1006/jmcc.1996.0366
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
We examined changes in the structural and physiological characteristics of human atrial myocytes during primary culture in the presence of serum. Action potentials and ionic currents were recorded in freshly dissociated (FM) and cultured (CM) whole-cell patch-clamped myocytes. alpha-smooth muscle actin, sarcomeric a-actinin and P-myosin heavy chains (beta-MHC) were stained with monoclonal antibodies. From day 5 to day 21, myocytes lost their rod shape, spread and exhibited reorganized sarcomeres. These morphological chances were associated with a marked increase in membrane capacitance (+266%). Both beta-MHC and a-smooth muscle actin were expressed in CM but not in FM, indicating a dedifferentiation process. CM were characterized by a lower resting potential (-30+/-2 v -60 +/- 4 mV, P<0.05) and, when repolarized, by a shorter action potential duration (APD) than FM (APD-60: 126.9 v 159.6 ms. P<0.05). The inward rectifier K+ current was absent in CM, thus explaining the low resting potential. The density of the transient component of the voltage-activated K+ current I-omega 1 was not modified during culture, while that of the sustained component I-SUS was increased fourfold. The amplitude of I-Ca was increased, but its density was unchanged, indicating that CM maintained a normal density of functional calcium channels. Neither the voltage dependence nor the inactivation of I-Ca was modified in CM. The time constants of inactivation of I-Ca were unchanged, although the amplitude of the rapidly inactivating component of I-Ca was increased in CM compared to FM. Moreover, I-Ca was increased by the beta-adrenergic agonist isoproterenol (1 mu M) throughout the culture period. Our results demonstrate that in long-term serum-supplemented culture, adaptation of human atrial myocytes to their new environment is associated with differential alterations of the main ionic currents and phenotypic changes characteristic of immature myocardium. (C) 1997 Academic Press Limited.
引用
收藏
页码:1307 / 1320
页数:14
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