Pfyffer G E, Bologa L, Herschkowitz N, Heizmann C W
J Neurochem. 1984 Jul;43(1):49-57. doi: 10.1111/j.1471-4159.1984.tb06677.x.
Dissociated brain cell cultures were derived from 14-day-old embryonic as well as from newborn mice. The cells were grown in a medium containing 10% fetal calf serum. Indirect immunofluorescence was performed using antisera directed against the Ca2+-binding protein parvalbumin (Mr 12,000). In embryonic cultures a large proportion of cells was intensely stained by antiparvalbumin . In double-labelling experiments involving the simultaneous application of antisera against parvalbumin and the neuron-specific enolase, the enolase-containing cells were also parvalbumin-positive and both antisera revealed identical intracellular staining patterns. Conversely, almost no parvalbumin- and enolase-positive cells were present in cultures derived from newborn mice. However, in these cultures many cells were immunoreactive toward the myelin basic protein, an accepted marker for oligodendrocytes. The presence of parvalbumin within the embryonic brain cell cultures was confirmed by analyses of the culture extracts (4 mM EDTA, pH 7.5) by HPLC on reverse-phase supports, two-dimensional polyacrylamide gel electrophoresis, and immunoblotting. The present study suggests that in mouse brain cell cultures, parvalbumin is localized in neurons.
解离的脑细胞培养物来源于14日龄胚胎小鼠以及新生小鼠。细胞在含有10%胎牛血清的培养基中生长。使用针对钙结合蛋白小白蛋白（分子量12,000）的抗血清进行间接免疫荧光。在胚胎培养物中，很大一部分细胞被抗小白蛋白强烈染色。在涉及同时应用抗小白蛋白和神经元特异性烯醇化酶抗血清的双重标记实验中，含烯醇化酶的细胞也是小白蛋白阳性，且两种抗血清显示出相同的细胞内染色模式。相反，新生小鼠来源的培养物中几乎不存在小白蛋白和烯醇化酶阳性细胞。然而，在这些培养物中，许多细胞对髓鞘碱性蛋白呈免疫反应性，髓鞘碱性蛋白是少突胶质细胞公认的标志物。通过在反相支持物上进行HPLC、二维聚丙烯酰胺凝胶电泳和免疫印迹分析培养提取物（4 mM EDTA，pH 7.5），证实了胚胎脑细胞培养物中存在小白蛋白。本研究表明，在小鼠脑细胞培养物中，小白蛋白定位于神经元。
