Thiel G, MacRobbie E A, Hanke D E
Botany School, University of Cambridge, UK.
EMBO J. 1990 Jun;9(6):1737-41. doi: 10.1002/j.1460-2075.1990.tb08297.x.
Inositol 1,4,5-trisphosphate (InsP3) was introduced into the cytoplasm of characean algae in two different ways: (i) by iontophoretic injection into cytoplasm-enriched fragments from Chara and (ii) by adding InsP3 to the permeabilization medium of locally permeabilized cells of Nitella. In both systems this operation induced a depolarization of the membrane potential, ranging from a few mV to sequences of action potentials. The effect of InsP3 on locally permeabilized Nitella cells was abolished when InsP3 was added together with 30 mM EGTA. When inositol 1,4-bisphosphate or myo-inositol were substituted for InsP3 in this system, there was no change in the membrane potential. On the other hand, increasing the free Ca2+ concentration in the permeabilization medium induced, in a similar fashion to InsP3, action potentials. Similarities between InsP3 and Ca2+ action were also observed upon injection into Chara fragments. Both injections increased an inward current. In the first few seconds after injection the current/voltage characteristics of the InsP3-induced current resembled those of the Ca2(+)-sensitive current. Subsequently, differences between the InsP3- and Ca2(+)-induced phenomena became apparent in that the InsP3-induced current continued to increase while the Ca2(+)-induced current declined, returning to the resting level. Our results suggest that these plant cells contain an InsP3 sensitive system that, under experimental conditions, is able to affect membrane transport via an increase in cytoplasmic free Ca2+.
肌醇1,4,5 -三磷酸(InsP3)通过两种不同方式被引入轮藻细胞的细胞质中:(i)通过离子电渗注入取自轮藻富含细胞质的片段;(ii)通过将InsP3添加到局部通透的丽藻细胞的通透介质中。在这两种体系中,此操作均诱导了膜电位的去极化,范围从几毫伏到动作电位序列。当InsP3与30 mM乙二醇双四乙酸(EGTA)一起添加时,InsP3对局部通透的丽藻细胞的作用被消除。当在此体系中用肌醇1,4 -二磷酸或肌醇替代InsP3时,膜电位没有变化。另一方面,增加通透介质中的游离Ca2 +浓度,以与InsP3类似的方式诱导动作电位。将InsP3和Ca2 +注入轮藻片段时也观察到了相似之处。两种注入均增加了内向电流。在注入后的最初几秒内,InsP3诱导电流的电流/电压特性类似于Ca2(+)敏感电流的特性。随后,InsP3和Ca2(+)诱导现象之间的差异变得明显,即InsP3诱导的电流持续增加,而Ca2(+)诱导的电流下降,恢复到静息水平。我们的结果表明,这些植物细胞含有一个InsP3敏感系统,在实验条件下,该系统能够通过增加细胞质游离Ca2 +来影响膜运输。