Yamamoto A, Haraguchi M, Yamashiro S, Fukumoto S, Furukawa K, Takamiya K, Atsuta M, Shiku H, Furukawa K
Department of Prosthodontology, Nagasaki University School of Dentistry, Japan.
J Neurochem. 1996 Jan;66(1):26-34. doi: 10.1046/j.1471-4159.1996.66010026.x.
Gangliosides are synthesized by sequential catalytic reaction of multiple glycosyltransferases. GM2/GD2 synthase and GD3 synthase are key enzymes for ganglioside synthesis, because their relative activities regulate the main profiles of ganglioside expression. Mouse GD3 synthase (EC 2.4.99.8) cDNA was cloned by eukaryotic expression cloning, and its mRNA expression as well as that of GM2/GD2 synthase gene during the development of the mouse CNS was analyzed by using northern blotting, reverse transcription-polymerase chain reaction, and in situ hybridization. When brain tissue was analyzed as a whole mass, a typical pattern corresponding to the reported findings obtained by biochemical analyses was observed, i.e., high expression of GD3 synthase gene in the early stage and gradual increase of GM2/GD2 synthase gene expression in the late stage of the development. However, the results of in situ hybridization of these two genes revealed that the expression kinetics of these two genes were heterogeneous among various sites in the brain under development. These findings suggest that various expression patterns of the two genes reflect differences in the course of the development of individual sites, and also different ganglioside components are required in individual portions of the brain for development and maintenance of the function.
神经节苷脂是通过多种糖基转移酶的顺序催化反应合成的。GM2/GD2合酶和GD3合酶是神经节苷脂合成的关键酶,因为它们的相对活性调节神经节苷脂表达的主要模式。小鼠GD3合酶(EC 2.4.99.8)cDNA通过真核表达克隆法克隆,并利用Northern印迹法、逆转录-聚合酶链反应和原位杂交技术分析了其mRNA表达以及GM2/GD2合酶基因在小鼠中枢神经系统发育过程中的表达。当将脑组织作为一个整体进行分析时,观察到一种与通过生化分析获得的报道结果相对应的典型模式,即在发育早期GD3合酶基因高表达,而在发育后期GM2/GD2合酶基因表达逐渐增加。然而,这两个基因的原位杂交结果显示,在发育中的大脑不同部位,这两个基因的表达动力学是异质的。这些发现表明,这两个基因的各种表达模式反映了各个部位发育过程的差异,并且大脑的各个部分在发育和维持功能方面需要不同的神经节苷脂成分。