Ito J I, Young Z L, Masuda-Isobe M, Tanaka R
Department of Biochemistry, Nagoya City University Medical School Mizuho-ku, Japan.
Neurochem Int. 1997 Oct;31(4):525-31. doi: 10.1016/s0197-0186(97)00021-1.
This study investigated in vitro the effects of gangliosides on polymerization of either the depolymerized microfilament preparation (MF) or depolymerized glia filament preparation (GF) extracted separately from the crude cytoskeletal fraction of rat astrocytes. Gangliosides GM1, GM2 and GM3 markedly suppressed polymerization of both MF and GF. The concentration of GM1, GM2 or GM3 required to induce 50% inhibition of the polymerization of 7.5 micrograms MF protein/200 microliters (IC50 of GM1, GM2, or GM3) was 3.2, 2.8 or 5.6 micrograms/200 microliters, respectively. The IC50 of each ganglioside for the polymerization of 7.5 micrograms/200 microliters of GF, furthermore, was 3.3, 3.5 or 7.4 micrograms/200 microliters, respectively, suggesting that the inhibitory activities of GM1 and GM2 on polymerization of both MF and GF were greater than those of GM3. GM1, GM2 and GM3 also suppressed dose-dependently the polymerization of both actin and vimentin. The inhibitory activities of GM1 and GM2 on the polymerization of actin or vimentin were greater than GM3, as in the case of polymerization of MF or GF. The IC50S of GD1a and GT1b for MF polymerization at the same concentration were 2.2 and 1.2 micrograms/200 microliters, respectively, and those for GF polymerization were 2.7 and 1.7 micrograms/200 microliters, respectively. The IC50 of GD3 for MF polymerization was 3.9 micrograms/200 microliters, and that for GF polymerization 4.0 micrograms/200 microliters, implying that the inhibitory activities of GD3 on polymerization of both MF and GF were greater than those of GM3. The findings suggested that the inhibitory activities of gangliosides on MF or GF polymerization became greater with increasing number of sialic acid residues. AsialoGM1 suppressed neither MF nor GF polymerization, and inhibited dose-dependently the ability of GM1 to suppress MF polymerization.
本研究在体外研究了神经节苷脂对分别从大鼠星形胶质细胞粗细胞骨架组分中提取的解聚微丝制剂(MF)或解聚神经胶质丝制剂(GF)聚合的影响。神经节苷脂GM1、GM2和GM3显著抑制MF和GF的聚合。诱导7.5微克MF蛋白/200微升聚合反应50%抑制所需的GM1、GM2或GM3浓度(GM1、GM2或GM3的IC50)分别为3.2、2.8或5.6微克/200微升。此外,每种神经节苷脂对7.5微克/200微升GF聚合的IC50分别为3.3、3.5或7.4微克/200微升,这表明GM1和GM2对MF和GF聚合的抑制活性大于GM3。GM1、GM2和GM3也剂量依赖性地抑制肌动蛋白和波形蛋白的聚合。GM1和GM2对肌动蛋白或波形蛋白聚合的抑制活性大于GM3,与MF或GF聚合的情况相同。相同浓度下,GD1a和GT1b对MF聚合的IC50分别为2.2和1.2微克/200微升,对GF聚合的IC50分别为2.7和1.7微克/200微升。GD3对MF聚合的IC50为3.9微克/200微升,对GF聚合的IC50为4.0微克/200微升,这意味着GD3对MF和GF聚合的抑制活性大于GM3。研究结果表明,神经节苷脂对MF或GF聚合的抑制活性随着唾液酸残基数量的增加而增强。去唾液酸GM1既不抑制MF也不抑制GF聚合,并剂量依赖性地抑制GM1抑制MF聚合的能力。