Doerner D, Pitler T A, Alger B E
Department of Physiology, University of Maryland School of Medicine, Baltimore 21201.
J Neurosci. 1988 Nov;8(11):4069-78. doi: 10.1523/JNEUROSCI.08-11-04069.1988.
Whole-cell voltage-clamp techniques were used to study the effects of the protein kinase C (PKC) activators phorbol esters and OAG on Ca and K currents in differentiated neurons acutely dissociated from adult hippocampus and in tissue-cultured neurons from fetal hippocampus. PKC activators had selective depressant effects on K currents, with persistent currents (IK and IK-Ca) being reduced and transient current (IA) being unaffected. In both cell types we recorded both high-voltage-activated, noninactivating (L-type) and high-voltage-activated, rapidly inactivating (N-type) Ca current. A low-voltage-activated, rapidly inactivating (T-type) Ca current was also recorded in tissue-cultured neurons but not in acutely dissociated neurons. PKC activators markedly reduced N-type current with less effect on L-type and no effect on T-type Ca current. Effects of PKC activators could be reversed with washing or with application of PKC inhibitors H-7 or polymyxin-B, an effect that could not be attributed to inhibition of cAMP-dependent protein kinase. The Ca/calmodulin inhibitor calmidazolium was ineffective in reversing the actions of PKC activators. Using whole-cell voltage-clamp techniques, we have demonstrated that hippocampal neurons possess 3 distinguishable components of calcium current. Distinct K currents were also observed. Our data strongly support the hypothesis that both Ca and K currents are selectively regulated by PKC and that these effects occur directly on the postsynaptic neuron.
采用全细胞电压钳技术研究蛋白激酶C(PKC)激活剂佛波酯和1-oleoyl-2-acetyl-sn-glycerol(OAG)对从成年海马急性分离的分化神经元以及胎儿海马组织培养神经元中钙电流和钾电流的影响。PKC激活剂对钾电流有选择性抑制作用,持续性电流(IK和IK-Ca)减少,而瞬时电流(IA)不受影响。在这两种细胞类型中,我们记录到了高电压激活的非失活(L型)钙电流和高电压激活的快速失活(N型)钙电流。在组织培养神经元中还记录到了低电压激活的快速失活(T型)钙电流,但在急性分离的神经元中未记录到。PKC激活剂显著降低N型电流,对L型电流影响较小,对T型钙电流无影响。PKC激活剂的作用可通过冲洗或应用PKC抑制剂H-7或多粘菌素B来逆转,这种作用不能归因于对环磷酸腺苷依赖性蛋白激酶的抑制。钙/钙调蛋白抑制剂氯咪唑对逆转PKC激活剂的作用无效。使用全细胞电压钳技术,我们证明海马神经元具有3种可区分的钙电流成分。还观察到了不同的钾电流。我们的数据有力地支持了以下假设:钙电流和钾电流均受PKC选择性调节,且这些作用直接发生在突触后神经元上。