Sumners C, Phillips M I, Raizada M K
Brain Res. 1983 Apr 4;264(2):267-75. doi: 10.1016/0006-8993(83)90824-7.
Catecholamines have been visualized and quantified in primary cultures of whole rat brain. Twenty-one-day old cultures treated with glyoxylic acid and viewed under a fluorescence microscope revealed neurons stained specifically with blue-green catecholamine fluorescence. Brightly stained multipolar cell bodies were seen, along with stained neurites and varicosities, and there was no staining associated with the non-neuronal portion of the culture. Twenty-one-day-old non-neuron-enriched cultures contained 10-20 times less norepinephrine and dopamine than cytosine arabinoside-treated neuron-enriched cultures. The latter cultures contained 10-12 times more norepinephrine than 1-day-old rat brains, demonstrating maturation and differentiation of the cultured neurons. Norepinephrine levels of neuron-enriched cultures were, however, 3 times less than those in 21-day-old rat brains. The cultured neurons had the ability to synthesize catecholamines since levels were decreased with alpha-methyl-p-tryosine. On the other hand, the growth medium contained significant amounts of norepinephrine, but did not have the ability to synthesize catecholamines. It may be concluded that the cellular catecholamines are not derived from the medium in any great amounts. This study provides the basis of a system in which to examine catecholaminergic neurotransmission and peptide catecholamine interactions at the cellular level under semi-defined conditions.
已在全大鼠脑原代培养物中对儿茶酚胺进行了可视化和定量分析。用乙醛酸处理的21日龄培养物在荧光显微镜下观察，可见神经元被蓝绿色儿茶酚胺荧光特异性染色。可见染色明亮的多极细胞体，以及染色的神经突和膨体，且培养物的非神经元部分无染色。21日龄非神经元富集培养物中的去甲肾上腺素和多巴胺含量比经阿糖胞苷处理的神经元富集培养物少10 - 20倍。后一种培养物中的去甲肾上腺素含量比1日龄大鼠脑多10 - 12倍，表明培养的神经元已成熟并分化。然而，神经元富集培养物中的去甲肾上腺素水平比21日龄大鼠脑低3倍。培养的神经元具有合成儿茶酚胺的能力，因为其含量会因α-甲基-对-酪氨酸而降低。另一方面，生长培养基中含有大量去甲肾上腺素，但没有合成儿茶酚胺的能力。可以得出结论，细胞内的儿茶酚胺并非大量来源于培养基。本研究为在半限定条件下在细胞水平研究儿茶酚胺能神经传递和肽-儿茶酚胺相互作用提供了一个系统基础。
