Neurons dissociated from rat myenteric plexus retain differentiated properties when grown in cell culture. III. Synaptic interactions and modulatory effects of neurotransmitter candidates.
Neurons dissociated from rat myenteric plexus retain differentiated properties when grown in cell culture. III. Synaptic interactions and modulatory effects of neurotransmitter candidates.
复制标题
从大鼠肌间神经丛分离的神经元在细胞培养物中生长时保留分化的特性。
DOI：
10.1016/0306-4522(85)90058-2
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发表时间：
1985
期刊：
影响因子：
3.3
通讯作者：
Nishi,R
中科院分区：
文献类型：
--
作者：
Willard,AL;Nishi,R
我们已经使用细胞内记录来研究在分离的细胞培养中生长的肌间神经元之间的突触相互作用。细胞内刺激单个肌间神经元可引起邻近神经元的几种类型的突触效应：（1）由烟碱乙酰胆碱受体介导的快兴奋性突触电位，（2）慢非胆碱能突触电位，（3）由烟碱乙酰胆碱受体介导的慢兴奋性突触电位。（3）具有快胆碱能和慢非胆碱能成分的双重传递，以及（4）抑制自发发生的快速烟碱突触电位快速烟碱突触电位由约40%的测试神经元引起，并且经常自发发生。快突触电位与其他自主神经元中研究的那些相似，它们的估计逆转电位和对胆碱能拮抗剂的敏感性。如果诱发频率大于0.5 Hz，则快突触电位的幅度下降。高频刺激突触前神经元后，可观察到快突触电位的增强。P物质和血管活性肠肽通过引起诱发和自发快突触电位的幅度增加和自发突触电位的频率增加来促进烟碱传递。γ-氨基丁酸和[Met]脑啡肽均引起烟碱突触电位的幅度和频率降低。5-羟色胺抑制某些神经元的突触电位，而增强它们或在其他神经元中没有影响。约10%的测试神经元引起缓慢的、非胆碱能的突触电位。这些突触效应持续15-300 s，引起3-15 mv的去极化，并伴有神经元输入阻力增加。当以10-20 Hz频率刺激时，一小部分胆碱能神经元在诱发快烟碱电位的同一突触后细胞中引起慢非胆碱能突触电位。这些5-12-mv的去极化持续数十秒，即使是非常高浓度的烟碱和毒蕈碱拮抗剂也不会减少。引起这些现象的递质还没有被确定，在培养的神经元之间观察到的唯一抑制性相互作用是间接的。当在5-20 Hz刺激时，约5%的测试细胞引起附近neurons.We的结论，在解离的细胞培养物中生长的肌间神经元可以形成几种类型的突触连接的自发发生的烟碱突触potentials的减少或停止。这些突触与体内肌间神经元中观察到的突触具有许多共同特性，包括被肠神经递质候选物调节的能力。因此，肌间神经元的解离细胞培养物将允许详细的生物物理学和药理学研究肠道神经传递及其调制。
We have used intracellular recordings to study synaptic interactions between myenteric neurons grown in dissociated cell culture. Intracellular stimulation of individual myenteric neurons caused several types of synaptic effects in nearby neurons: (1) fast excitatory synaptic potentials mediated by nicotinic acetylcholine receptors; (2) slow, non-cholinergic synaptic potentials; (3) dual transmission having both fast cholinergic and slow non-cholinergic components and (4) inhibition of spontaneously occurring fast nicotinic synaptic potentials.Fast nicotinic synaptic potentials were elicited by about 40% of neurons tested and often occurred spontaneously. The fast synaptic potentials were similar to those that have been studied in other autonomic neurons with respect to their estimated reversal potential and their sensitivity to cholinergic antagonists. The amplitudes of the fast synaptic potentials declined if evoked at frequencies greater than 0.5 Hz. Potentiation of the fast synaptic potentials was observed following high-frequency stimulation of presynaptic neurons.Several transmitter candidates modulated fast cholinergic transmission. Substance P and vasoactive intestinal peptide promoted nicotinic transmission by causing increased amplitudes of evoked and spontaneous fast synaptic potentials and an increased frequency of spontaneous synaptic potentials. γ-Aminobutyrate and [Met]enkephalin both caused decreased amplitudes and frequency of nicotinic synaptic potentials. Serotonin depressed synaptic potentials in some neurons while enhancing them or having no effect in others.Slow, non-cholinergic, synaptic potentials were elicited by about 10% of neurons tested. These synaptic effects lasted 15–300 s, caused depolarizations of 3–15 mv and were accompanied by increased neuronal input resistance. The transmitter(s) causing these slow synaptic potentials has not yet been identified.When stimulated at 10–20 Hz, a small proportion of cholinergic neurons caused slow non-cholinergic synaptic potentials in the same postsynaptic cells in which they evoked fast nicotinic potentials. These 5–12-mv depolarizations lasted tens of seconds and were not reduced even by very high concentrations of both nicotinic and muscarinic antagonists. The transmitters causing them have not yet been identified.The only type of inhibitory interaction observed between the cultured neurons was an indirect one. When stimulated at 5–20 Hz, about 5% of cells tested caused a diminution or cessation of spontaneously occurring nicotinic synaptic potentials in nearby neurons.We conclude that myenteric neurons grown in dissociated cell cultures can form several types of synaptic connections. These synapses share many properties with those that have been observed in myenteric neuronsin vivo, including the ability to be modulated by enteric neurotransmitter candidates. Thus dissociated cell cultures of myenteric neurons will permit detailed biophysical and pharmacological studies of enteric neurotransmission and of its modulation.