Dzhura I, Kostyuk P, Lyubanova O, Naidenov V, Shuba Y
Bogomoletz Institute of Physiology, National Academy of Sciences of Ukraine, Kiev.
Neuroreport. 1994 Oct 3;5(15):1960-2. doi: 10.1097/00001756-199410000-00030.
Xenopus laevis oocytes were injected with total mRNA obtained from the thalamo-hypothalamic complex of adult rats. In 19 out of 32 injected oocytes a Ba2+ current was expressed after 4 days which could be activated at depolarizations to -70 mV from a holding potential of -120 mV and reached a maximum value at between -30 and -20 mV. The current inactivated monoexponentially with a time constant of about 420 +/- 10 ms (n = 6); its steady state inactivation had a half value of -78 +/- 1 mV (n = 8) and a slope (k) of 11.5 +/- 3.0. These characteristics are typical of LVA (T-type) Ca2+ channels in neurones from the corresponding brain structures, except for the much slower time course of inactivation. These currents were blocked by pharmacological antagonists specific for LVA channels (amiloride, flunarizine), but remained resistant to omega-Aga-IVA and omega-Cg-toxin. These results show that LVA Ca2+ channels can be expressed in oocytes provided that the corresponding mRNA is taken from brain neurones in which they are naturally well expressed.
将成年大鼠丘脑 - 下丘脑复合体获得的总mRNA注射到非洲爪蟾卵母细胞中。在32个注射的卵母细胞中,有19个在4天后表达出Ba2 +电流,该电流可在从 - 120 mV的钳制电位去极化到 - 70 mV时被激活,并在 - 30至 - 20 mV之间达到最大值。电流以单指数形式失活,时间常数约为420±10 ms(n = 6);其稳态失活的半值为 - 78±1 mV(n = 8),斜率(k)为11.5±3.0。除了失活的时间进程要慢得多之外,这些特征是来自相应脑结构的神经元中低电压激活(T型)Ca2 +通道的典型特征。这些电流被对低电压激活通道特异的药理拮抗剂(氨氯吡脒、氟桂利嗪)阻断,但对ω - 芋螺毒素IVA和ω - 海葵毒素仍有抗性。这些结果表明,只要相应的mRNA取自天然表达低电压激活Ca2 +通道的脑神经元,低电压激活Ca2 +通道就可以在卵母细胞中表达。