Gallo V, Balazs R, Jørgensen O S
Brain Res. 1985 Jan;349(1-2):27-37. doi: 10.1016/0165-3806(85)90129-4.
Lactoperoxidase catalysed 125I-iodination of cerebellar interneurone enriched cultures grown in serum-supplemented or in serum-free, chemically defined medium was studied. It was observed that the differences in the adhesion properties of nerve cells under these conditions are accompanied by differences both in the degree of 125I-iodination of the proteins on the plasma membrane of nerve cells and in the profile of the labelled polypeptides resolved by SDS-polyacrylamide gel electrophoresis. The relative labelling of the major 125I-iodinated polypeptides changed with time in both types of cultures, suggesting that alterations in the overall organisation of the neuronal plasma membrane occur during the development of the cells under both culture conditions. The developmental changes affecting the D2 protein (which is nerve cell specific in these cultures) were significantly retarded in the neuronal cultures grown in the serum-free medium compared with those grown in the serum-containing medium: the increase in D2 content was reduced by 7 DIV and the maturational change in the molecular form of D2 was retarded significantly during the whole cultivation period. The degree of surface 125I-iodination of cerebellar astrocytes in culture was only a fraction (7-20% depending on cultivation time and conditions) of that of the neuronal cultures and the labelled polypeptide profiles obtained from the two types of cultures were markedly different. In comparison with cultures in the serum-supplemented medium, astrocytes under the serum-free conditions showed only minor and transient differences in the profile of surface 125I-iodinated proteins, although the morphology of the cells was markedly different.
研究了乳过氧化物酶催化的在补充血清或无血清、化学成分明确的培养基中生长的富含小脑中间神经元的培养物的125I碘化。观察到在这些条件下神经细胞黏附特性的差异伴随着神经细胞质膜上蛋白质的125I碘化程度以及通过SDS-聚丙烯酰胺凝胶电泳分离的标记多肽谱的差异。在两种类型的培养物中,主要的125I碘化多肽的相对标记随时间变化,这表明在两种培养条件下细胞发育过程中神经元质膜的整体组织发生了改变。与在含血清培养基中生长的神经元培养物相比,在无血清培养基中生长的神经元培养物中影响D2蛋白(在这些培养物中是神经细胞特异性的)的发育变化明显延迟:D2含量的增加在第7天减少,并且在整个培养期间D2分子形式的成熟变化明显延迟。培养的小脑星形胶质细胞表面125I碘化程度仅为神经元培养物的一小部分(取决于培养时间和条件,为7%-20%),并且从两种类型的培养物中获得的标记多肽谱明显不同。与补充血清的培养基中的培养物相比,无血清条件下的星形胶质细胞在表面125I碘化蛋白质谱中仅表现出微小的短暂差异,尽管细胞形态明显不同。