Gomez J P, Potreau D, Branka J E, Raymond G
Laboratory of General Physiology, URA C.N.R.S. 1869, Faculty of Sciences, Poitiers, France.
Pflugers Arch. 1994 Oct;428(3-4):241-9. doi: 10.1007/BF00724503.
Electrophysiological characteristics of neonatal rat ventricular cardiomyocytes in primary culture were studied using the whole-cell patch-clamp recording technique. Cell size, estimated by measurement of membrane capacitance, was significantly increased throughout the culture from 22.4 +/- 5.4 pF at day 2 to 55.0 +/- 16.1 pF at day 7, reflecting the hypertrophic process which characterises postnatal cell development. The Ca2+ current was investigated at day 2 and 7 of the culture which constituted the early postnatal and maximally developed stages, respectively, of isolated cells in our experimental conditions. At 2 days of culture, two types of Ca2+ current could be distinguished, as also observed in freshly dissociated newborn ventricular cells. From their potential dependence and pharmacological characteristics, they could be attributed to the T- (ICa-T) and L-type (ICa-L) Ca2+ current components. After 7 days of culture, only the latter ICa-L was present and its density was significantly increased when compared to the density in 2-day-old cells, but lower than that obtained in freshly dissociated adult cells. As the age of the culture progressed, the steady-state inactivation curve was shifted toward negative potentials, in the direction of the inactivation curve obtained for adult cells. Compared to the serum-free control conditions, the density of ICa-L was significantly increased in the presence of fetal calf serum throughout the culture. Consequently, the density of ICa-L obtained in 7-day-old cells was similar to the density of ICa-L obtained in freshly dissociated adult cardiac cells.(ABSTRACT TRUNCATED AT 250 WORDS)
采用全细胞膜片钳记录技术,研究了原代培养新生大鼠心室心肌细胞的电生理特性。通过测量膜电容估算细胞大小,整个培养过程中细胞大小显著增加,从第2天的22.4±5.4 pF增加到第7天的55.0±16.1 pF,反映了出生后细胞发育的肥厚过程。在培养的第2天和第7天研究了Ca2+电流,在我们的实验条件下,这分别代表了分离细胞出生后的早期和最大发育阶段。在培养2天时,可以区分出两种类型的Ca2+电流,这在刚分离的新生心室细胞中也有观察到。根据它们的电位依赖性和药理学特性,可将其归因于T型(ICa-T)和L型(ICa-L)Ca2+电流成分。培养7天后,仅存在后者ICa-L,与2日龄细胞相比,其密度显著增加,但低于刚分离的成年细胞。随着培养时间的延长,稳态失活曲线向负电位移动,与成年细胞的失活曲线方向一致。与无血清对照条件相比,在整个培养过程中,胎牛血清存在时ICa-L的密度显著增加。因此,7日龄细胞中获得的ICa-L密度与刚分离的成年心脏细胞中获得的ICa-L密度相似。(摘要截断于250字)