Ferret B, Hubsch A, Dreyfus H, Massarelli R
Centre de Neurochimie du CNRS, Strasbourg, France.
Neurochem Res. 1991 Feb;16(2):137-44. doi: 10.1007/BF00965701.
Primary neurons in culture from chick embryo cerebral hemispheres were treated with a mixture of gangliosides added to the growth medium (final concentration: 10(-5)M and 10(-8)M) from the 3rd to the 6th day in vitro. Under these conditions methylation processes measured with [3H] and [35S] methionine and [3H]ethanolamine as precursors showed an increased methylation of [3H]ethanolamine containing phospholipids, a correspondent increased conversion of these compounds to [3H]choline containing phospholipids, and a general increased methylation of trichloroacetic acid precipitable macromolecules containing labeled methionine. A small increase in protein synthesis was observed after incubation of neurons with [3H]- and [35S]methionine. This was confirmed after electrophoretic separation of a protein extract with increased 3H- and 35S-labeling in protein bands with moecular weights between 50 and 60 KDaltons. A protein band of about 55 KDaltons appeared to be preferentially labelled when [3H] methionine was the precursor. The treatment with gangliosides increased the incorporation of [methyl-3H] label after incubation of neurons with [3H] methionine, into total DNA and decreased that of total RNA. The treatment of neurons in culture with exogenous gangliosides hence affects differently methylation processes, a finding which may confirm the involvement of gangliosides on the intracellular mediation of neuronal information mechanisms.
将鸡胚大脑半球培养的原代神经元,在体外培养的第3天至第6天,用添加到生长培养基中的神经节苷脂混合物处理(终浓度:10⁻⁵M和10⁻⁸M)。在这些条件下,以[³H]和[³⁵S]甲硫氨酸以及[³H]乙醇胺作为前体测量的甲基化过程显示,含[³H]乙醇胺的磷脂甲基化增加,这些化合物向含[³H]胆碱的磷脂的相应转化率增加,并且含标记甲硫氨酸的三氯乙酸可沉淀大分子的总体甲基化增加。在用[³H] - 和[³⁵S]甲硫氨酸孵育神经元后,观察到蛋白质合成略有增加。在用分子量在50至60千道尔顿之间的蛋白质条带中具有增加的³H - 和³⁵S标记的蛋白质提取物进行电泳分离后,这一点得到了证实。当以[³H]甲硫氨酸作为前体时,一条约55千道尔顿的蛋白质条带似乎被优先标记。在用[³H]甲硫氨酸孵育神经元后,神经节苷脂处理增加了[甲基 - ³H]标记掺入总DNA中的量,并减少了总RNA中的掺入量。因此,用外源性神经节苷脂处理培养的神经元会对甲基化过程产生不同的影响,这一发现可能证实了神经节苷脂参与神经元信息机制的细胞内介导。