Mains R E, Patterson P H
J Cell Biol. 1973 Nov;59(2 Pt 1):329-45. doi: 10.1083/jcb.59.2.329.
Rat sympathetic ganglia were disrupted by mechanical agitation to yield dissociated primary neurons, and the conditions for long-term growth in culture of the isolated neurons were examined. The neurons were grown with or without non-neural cells, simply by the addition or deletion of bicarbonate during growth in culture. Fluorescence histochemistry indicated that the isolated neurons contained catecholamines; incubations with radioactive precursors were used to verify the synthesis and accumulation of both dopamine and norepinephrine. The neurons also produced octopamine using tyramine as precursor, but not with tyrosine as the precursor. In the presence of eserine, older cultures synthesized and stored small amounts of acetylcholine. The cultures did not synthesize and accumulate detectable levels of radioactive gamma-aminobutyric acid, 5-hydroxytryptamine, or histamine.
通过机械搅拌破坏大鼠交感神经节以获得解离的原代神经元,并研究分离出的神经元在培养中长期生长的条件。在培养过程中,通过简单地添加或去除碳酸氢盐,使神经元在有或没有非神经细胞的情况下生长。荧光组织化学表明,分离出的神经元含有儿茶酚胺;使用放射性前体进行孵育以验证多巴胺和去甲肾上腺素的合成与积累。这些神经元还利用酪胺作为前体产生章鱼胺,但不能利用酪氨酸作为前体。在存在毒扁豆碱的情况下,较老的培养物合成并储存少量乙酰胆碱。这些培养物不会合成并积累可检测水平的放射性γ-氨基丁酸、5-羟色胺或组胺。