Fu W M, Liou H C, Chen Y H
Pharmacological Institute, College of Medicine, National Taiwan University, Taipei, Taiwan 100.
J Neurosci. 1998 Dec 1;18(23):9954-61. doi: 10.1523/JNEUROSCI.18-23-09954.1998.
The activation of autoreceptors is known to be important in the modulation of presynaptic transmitter secretion in peripheral and central neurons. Using whole-cell recordings made from the free growth cone of myocyte-contact motoneurons of Xenopus cell cultures, we have observed spontaneous nerve terminal currents (NTCs). These spontaneous NTCs are blocked by d-tubocurarine (d-TC) and alpha-bungarotoxin (alpha-BuTx), indicating that endogenously released acetylcholine (ACh) can produce substantial membrane depolarization in the nerve terminals. Local application of NMDA to the growth cone increased the frequency of spontaneous NTCs. When the electrical stimulations were applied at the soma to initiate evoked-release of ACh, evoked ACh-induced potentials were recorded in the nerve terminals, which were inhibited by d-TC and hexamethonium but not by atropine. Replacement of normal Ringer's solution with high-Mg2+, low-Ca2+ solution also reversibly inhibited evoked ACh-induced potentials. The possible regulatory role of presynaptic nicotinic autoreceptors on the synaptic transmission was also examined. When the innervated myocyte was whole-cell voltage-clamped to record synaptic currents, application of hexamethonium inhibited the amplitude of evoked synaptic currents at a higher degree than that of iontophoretic ACh-induced currents. Furthermore, hexamethonium markedly reduced the frequency of spontaneous synaptic currents at high-activity synapses. Pretreatment of neurons with alpha-BuTx also inhibited the evoked synaptic currents in manipulated synapses. These results suggest that ACh released spontaneously or by electrical stimulation may act on the presynaptic nicotinic autoreceptors of the same nerve terminals to produce membrane potential change and to regulate synaptic transmission.
已知自身受体的激活在调节外周和中枢神经元的突触前递质分泌中起着重要作用。利用非洲爪蟾细胞培养物中与肌细胞接触的运动神经元自由生长锥进行的全细胞记录,我们观察到了自发神经末梢电流(NTCs)。这些自发的NTCs被d -筒箭毒碱(d - TC)和α -银环蛇毒素(α - BuTx)阻断,表明内源性释放的乙酰胆碱(ACh)可在神经末梢产生显著的膜去极化。将NMDA局部应用于生长锥可增加自发NTCs的频率。当在胞体施加电刺激以引发ACh的诱发释放时,在神经末梢记录到诱发的ACh诱导电位,该电位被d - TC和六甲铵抑制,但不被阿托品抑制。用高镁、低钙溶液替代正常的林格氏液也可可逆地抑制诱发的ACh诱导电位。还研究了突触前烟碱型自身受体对突触传递的可能调节作用。当对受支配的肌细胞进行全细胞电压钳制以记录突触电流时,应用六甲铵对诱发突触电流幅度的抑制程度高于对离子电泳ACh诱导电流的抑制程度。此外,六甲铵在高活性突触处显著降低了自发突触电流的频率。用α - BuTx预处理神经元也抑制了被操作突触中的诱发突触电流。这些结果表明,自发释放或通过电刺激释放的ACh可能作用于同一神经末梢的突触前烟碱型自身受体,以产生膜电位变化并调节突触传递。