Chen C, Clarke I J
Prince Henry's Institute of Medical Research, Clayton, Victoria, Australia.
Am J Physiol. 1995 Feb;268(2 Pt 1):E204-12. doi: 10.1152/ajpendo.1995.268.2.E204.
Voltage-gated Ca2+ currents were recorded using the nystatin-perforated whole cell recording configuration on the ovine somatotrophs. With the use of Ca(2+)-tetraethylammonium chloride bath solution and Cs+ electrode solution, two types of Ca2+ currents were obtained with a predominant long-lasting (L) current blocked by nifedipine. A transient (T) current was isolated in the presence of nifedipine (3 microM) and was not blocked by omega-conotoxin (5 microM), but diminished to 47 +/- 5% of control by Ni2+ (0.3 mM) or to 52 +/- 10% of control by amiloride (0.5 mM). The nifedipine-blockable L-type current was not affected by omega-conotoxin (5 microM); it was, however, attenuated to 80 +/- 4% of control by Ni2+ (0.3 mM) and to 48 +/- 6% of control by amiloride (0.5 nM). Cd2+ (1 mM) totally prevented both T and L currents. Application of growth hormone-releasing factor (GRF, 10 nM) reversibly increased the amplitude of both Ca2+ currents without modifying their kinetic properties. The effect of GRF was observed approximately 30 s after application, peaked (142 +/- 11% of control, n = 5) rapidly, and lasted > 10 min if GRF treatment was continuous. Intracellular Ca2+ concentration ([Ca2+]i) was increased by GRF (10 nM) within seconds, reaching a peak within 30 s and lasting > 250 s. Blockade of Ca2+ channels (Cd2+, 1 mM) or the use of Ca(2+)-free solution reduced basal [Ca2+]i and significantly (P < 0.05) diminished the effect of GRF on [Ca2+]i.(ABSTRACT TRUNCATED AT 250 WORDS)
采用制霉菌素穿孔全细胞记录模式,在绵羊生长激素分泌细胞上记录电压门控性Ca2+电流。使用Ca(2+)-四乙铵氯化物灌流液和Cs+电极内液,可获得两种类型的Ca2+电流,其中一种主要的长时程(L)电流可被硝苯地平阻断。在存在硝苯地平(3 microM)的情况下可分离出瞬态(T)电流,该电流不被ω-芋螺毒素(5 microM)阻断,但被Ni2+(0.3 mM)降低至对照的47±5%,或被阿米洛利(0.5 mM)降低至对照的52±10%。可被硝苯地平阻断的L型电流不受ω-芋螺毒素(5 microM)影响;然而,它被Ni2+(0.3 mM)降低至对照的80±4%,被阿米洛利(0.5 nM)降低至对照的48±6%。Cd2+(1 mM)完全阻断T电流和L电流。应用生长激素释放因子(GRF,10 nM)可使两种Ca2+电流的幅度可逆性增加,而不改变其动力学特性。GRF的作用在应用后约30秒观察到,迅速达到峰值(对照的142±11%,n = 5),如果持续给予GRF,作用持续超过10分钟。细胞内Ca2+浓度([Ca2+]i)在数秒内被GRF(10 nM)升高,在30秒内达到峰值并持续超过250秒。阻断Ca2+通道(Cd2+,1 mM)或使用无Ca(2+)溶液可降低基础[Ca2+]i,并显著(P < 0.05)减弱GRF对[Ca2+]i的作用。(摘要截短于250字)