Meis S, Deitmer J W
Abteilung für Allgemeine Zoologie, FB Biologie, Universität Kaiserslautern, Germany.
J Neurosci Res. 1997 Dec 15;50(6):967-78. doi: 10.1002/(SICI)1097-4547(19971215)50:6<967::AID-JNR7>3.0.CO;2-B.
The expression of calcium-activated potassium currents (IK(Ca)), delayed outward rectifier potassium currents (IK(slow)), and transient outward currents (IA) was studied during the development of the nervous system of the leech using the whole-cell patch-clamp recording technique. Dissociated cells were isolated from leech embryos between stage E7 and E16 and maintained in primary culture. K+ currents were recorded at E7, when only few anterior ganglia had formed beneath the primordial mouth. IK(slow) was present in all cells tested, while IK(Ca) was expressed in only 67% of the cells studied. Even as early as E7, different types of IK(Ca) have been found. Neither frequency of occurrence nor the charge density of IK(Ca) showed significant changes between E7 and E16. The density of IK(slow), however, increased by a factor of two between E7 and E8, which resulted in a significant increase in the total K+ current of these cells. This rise in potassium outward current developed in parallel with the appearance of Na+ and Ca2+ inward currents (Schirrmacher and Deitmer: J Exp Biol 155:435-453, 1991) during early development, shaping the electrical excitability in embryonic leech neurones. I(A) could be separated by its voltage-dependence and pharmacological properties. The current was detected at stage E9, when all 32 ganglia are formed in the embryo. The frequency of occurrence of I(A) increased from 16% at E9 to 70% at E15. The channel density, steady state inactivation, and kinetics showed no significant changes during development.
利用全细胞膜片钳记录技术,研究了水蛭神经系统发育过程中钙激活钾电流(IK(Ca))、延迟外向整流钾电流(IK(slow))和瞬时外向电流(IA)的表达情况。从E7至E16期的水蛭胚胎中分离出解离细胞,并进行原代培养。在E7期记录钾电流,此时在原口下方仅形成了少数前神经节。在所测试的所有细胞中均存在IK(slow),而在所研究的细胞中只有67%表达IK(Ca)。甚至早在E7期,就已发现不同类型的IK(Ca)。在E7至E16期之间,IK(Ca)的出现频率和电荷密度均未显示出显著变化。然而,IK(slow)的密度在E7至E8期之间增加了两倍,这导致这些细胞的总钾电流显著增加。钾外向电流的这种增加与早期发育过程中钠和钙内向电流的出现同时发生(Schirrmacher和Deitmer:J Exp Biol 155:435 - 453, 1991),塑造了胚胎水蛭神经元的电兴奋性。IA电流可通过其电压依赖性和药理学特性进行分离。在E9期检测到该电流,此时胚胎中所有32个神经节均已形成。IA电流的出现频率从E9期的16%增加到E15期的70%。在发育过程中,通道密度、稳态失活和动力学均未显示出显著变化。