Neurons dissociated from rat myenteric plexus retain differentiated properties when grown in cell culture. I. Morphological properties and immunocytochemical localization of transmitter candidates.
Neurons dissociated from rat myenteric plexus retain differentiated properties when grown in cell culture. I. Morphological properties and immunocytochemical localization of transmitter 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)90056-9
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发表时间：
1985
期刊：
影响因子：
3.3
通讯作者：
Willard,AL
中科院分区：
文献类型：
--
作者：
Nishi,R;Willard,AL
我们已经开发了从新生大鼠的小肠肌间神经丛中分离神经元并在细胞培养中培养这些神经元长达3个月的程序。这些培养的神经元保留了许多肌间神经在体内的分化特性。这是描述这些特性的三篇系列论文中的第一篇。在本文中，我们描述了我们用光学和电子显微镜观察到的培养神经元的形态，也描述了免疫细胞化学技术在培养神经元中定位神经递质候选时观察到的应变模式。细胞内注射荧光染料荧光黄显示，许多培养的神经元具有与体内肌间神经元相似的形态。在电子显微镜下观察培养的薄片，可见许多神经元有许多小的(40-60 nm)透明的突触小泡和(或)大的(80-150 nm)不透明的核心(p型)小泡。含有5-羟色胺、P物质、生长抑素、脑啡肽、蛙黄素和胃泌素/缩胆囊素免疫反应阳性神经元的比例与完整的肌间神经丛大致相同。含有免疫活性血管活性肠多肽的神经元的数量比体内报道的要多。未见神经降压素、促胰液素和谷氨酸脱羧酶免疫反应阳性神经元。针对胆碱乙酰转移酶的抗血清染色了40%-50%的神经元。我们得出结论，即使从肠道中取出肌间神经元，分离成单个细胞的悬液并在培养中生长，肌间神经元仍继续表现出许多正常的分化特性。这种培养将有助于关联单个肌间神经元的形态、生物物理、药理学和突触属性，并测试改变的环境条件改变这些属性的能力。
We have developed procedures for dissociating neurons from the myenteric plexus of the small intestine of newborn rats and for growing those neurons in cell cultures for up to 3 months. Neurons in these cultures retain many of the differentiated properties of myenteric neuronsin vivo. This is the first of a series of 3 papers describing those properties. In this paper, we describe the morphology of cultured neurons that we have observed with light and electron microscopy; we also describe the patterns of straining observed when immunocytochemical techniques were used to localize neurotransmitter candidates in the cultured neurons.Intracellular injections of a fluorescent dye, Lucifer yellow, revealed that many of the cultured neurons had morphologies similar to those of myenteric neuronsin vivo. When thin sections of cultures were viewed in an electron microscope, many neurons were observed to have numerous small (40–60 nm), clear synaptic vesicles and/or large (80–150 nm), opaque-cored (p-type) vesicles. Synaptic profiles were most often observed on neuronal somata.Neurons containing immunoreactive serotonin, substance P, somatostatin, enkephalin, bombesin and gastrin/cholecystokinin were observed in about the same proportions as they occur in the intact myenteric plexus. Neurons containing immunoreactive vasoactive intestinal polypeptide were found in higher numbers than reportedin vivo. Neurons containing immunoreactive neurotensin, secretin and glutamate decarboxylase were not observed. An antiserum directed against choline acetyltransferase stained 40–50% of the neurons.We conclude that myenteric neurons continue to express much of their normal differentiated properties even when they are removed from the gut, dissociated into a suspension of single cells and grown in culture. Such cultures will be useful for correlating the morphological, biophysical, pharmacological and synaptic properties of individual myenteric neurons and for testing the ability of altered environmental conditions to change those properties.