Watanabe K, Oohira A, Katoh-Semba R, Totsuka T, Yoshida K
Department of Physiology, Aichi Prefecture Colony, Japan.
Neurochem Res. 1989 Aug;14(8):707-16. doi: 10.1007/BF00964947.
Mouse neuroblastoma Neuro 2a cells are known to extend neurite-like processes in response to gangliosides added to the culture medium. We compared the structural features of proteoglycans (PG) synthesized by conventional Neuro 2a cells with those of neurite-bearing cells. Two different proteoglycans labeled with [35S]sulfate, namely, chondroitin sulfate proteoglycan (CS-PG) and heparan sulfate proteoglycan (HS-PG), were found both in the cell layer and in the culture medium of the conventional cells. CS-PG isolated from the cell layer had a Kav value of 0.38 on Sepharose CL-6B, and had CS side chains with Mr of 27,000. HS-PG in the cell layer was slightly larger (Kav of 0.33) in terms of hydrodynamic size than CS-PG, and the apparent Mr of the heparan sulfate side chains was 10,000. The structural parameters of CS-PG and HS-PG isolated from the medium were almost identical to those of the PGs in the cell layer. In addition to these PGs, single-chain HS, with an average Mr of 2,500, was observed only in the cell layer and this component was the major sulfated component in the cell layers of both control and ganglioside treated cells. The neurite-bearing cells also synthesized both CS-PG and HS-PG which were very similar in hydrodynamic size to those synthesized by the conventional cells, but the size of HS side chains was greater. Radioactivity, as 35S, of each sulfated component from the ganglioside-treated culture seemed to be slightly less than that of the corresponding component from the control culture. These findings indicate that the marked morphological change in Neuro 2a cells, induced by gangliosides is not accompanied by major changes in the synthesis of PGs.
已知小鼠神经母细胞瘤Neuro 2a细胞会因向培养基中添加神经节苷脂而伸出类神经突样突起。我们比较了传统Neuro 2a细胞合成的蛋白聚糖(PG)与有神经突细胞的蛋白聚糖的结构特征。在传统细胞的细胞层和培养基中均发现了两种用[35S]硫酸盐标记的不同蛋白聚糖,即硫酸软骨素蛋白聚糖(CS-PG)和硫酸乙酰肝素蛋白聚糖(HS-PG)。从细胞层分离的CS-PG在琼脂糖CL-6B上的Kav值为0.38,其CS侧链的分子量为27,000。细胞层中的HS-PG在流体力学尺寸方面比CS-PG略大(Kav为0.33),硫酸乙酰肝素侧链的表观分子量为10,000。从培养基中分离的CS-PG和HS-PG的结构参数与细胞层中的PG几乎相同。除了这些PG外,仅在细胞层中观察到平均分子量为2500的单链HS,并且该成分是对照细胞和神经节苷脂处理细胞的细胞层中的主要硫酸化成分。有神经突的细胞也合成了CS-PG和HS-PG,它们在流体力学尺寸上与传统细胞合成的非常相似,但HS侧链的尺寸更大。神经节苷脂处理培养物中各硫酸化成分的35S放射性似乎略低于对照培养物中相应成分的放射性。这些发现表明,神经节苷脂诱导的Neuro 2a细胞明显的形态变化并未伴随PG合成的主要变化。