Chondroitin sulfate/dermatan sulfate hybrid chains in the development of cerebellum - Spatiotemporal regulation of the expression of critical disulfated disaccharides by specific sulfotransferases
Chondroitin sulfate/dermatan sulfate hybrid chains in the development of cerebellum - Spatiotemporal regulation of the expression of critical disulfated disaccharides by specific sulfotransferases
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DOI：
10.1074/jbc.m510870200
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发表时间：
2006-07-14
影响因子：
4.8
通讯作者：
Sugahara, Kazuyuki
中科院分区：
文献类型：
--
作者：
Mitsunaga, Chie;Mikami, Tadahisa;Sugahara, Kazuyuki
硫酸软骨素/硫酸皮肤素（CS/DS）链调节脊椎动物中枢神经系统的发育。以前，我们证明了胚胎猪脑的CS/DS杂交链表现出神经轴突和生长因子结合活性，这取决于它们的IdoUA含量定义的DS样结构。为了阐明这种功能性糖链在大脑发育过程中的分布，进行原位杂交以检查三种CS/DS GalNAc 4-O-磺基转移酶（D4 ST-1、C4 ST-1和C4 ST-2）和一种参与DS生物合成的糖醛酸2-O-磺基转移酶（UST）以及CS中间体的表达。C4 ST-1和C4 ST-2在出生后小鼠脑中普遍表达，而D4 ST-1和UST的表达仅限于发育中的小脑，并在出生后第14天达到高峰，如通过逆转录-PCR分析所示。在大脑切片中的CS/DS的二糖的原位分析表明，CS/DS的浓度增加2倍，在发展过程中（出生后第7天至7周）。由D4 ST-1和UST顺序作用产生的DS特异性主要二糖IdoUA-GalNAc（4-O-硫酸盐）（iA）和IdoUA（2-O-硫酸盐）-GalNAc（4-O-硫酸盐）（iB）的比例在小脑CS/DS链中高于全脑切片中的比例。值得注意的是，在发育过程中，iB的比例急剧增加（10倍）。相反，在发育中的小脑中，GlcUA/IdoUA（2-O-硫酸盐）-GalNAc（6-O-硫酸盐）（D/iD）和GlcUA/IdoUA-GalNAc（4，6-O-二硫酸盐）（E/iE）分别降低至50%和30%。这些结果表明，含IdoUA的iA和iB单位沿着D/iD和E/iE单位在出生后脑发育过程中对小脑神经网络的形成起重要作用。
Chondroitin sulfate/dermatan sulfate (CS/DS) chains regulate the development of the central nervous system in vertebrates. Previously, we demonstrated that CS/DS hybrid chains from embryonic pig brain exhibit neuritogenic and growth factor binding activities, which depended on their IdoUA content defining the DS-like structure. To elucidate the distribution of such functional sugar chains during the development of the brain, in situ hybridization was performed to examine expression of three CS/DS GalNAc 4-O-sulfotransferases, D4ST-1, C4ST-1, and C4ST-2, and a single uronyl 2-O-sulfotransferase (UST) involved in the biosynthesis of DS in addition to CS intermediates. C4ST-1 and C4ST-2 were ubiquitously expressed in the postnatal mouse brain, whereas the expression of D4ST-1 and UST was restricted in the developing cerebellum and culminated at postnatal day 14 as shown by reverse transcriptase-PCR analysis. In situ analysis of the disaccharides of CS/DS in brain sections revealed that the concentration of CS/DS increases 2-fold during development (postnatal day 7 to 7 weeks). The proportions of DS-specific, principal disaccharides, IdoUA-GalNAc(4-O-sulfate) (iA) and IdoUA(2-O-sulfate)-GalNAc(4-O-sulfate) (iB), produced by the sequential actions of D4ST-1 and UST, were higher in the CS/DS chains from cerebellum than those from whole brain sections. A dramatic increase (10-fold) in the proportion of iB during development was noteworthy. In contrast, GlcUA/IdoUA(2-O-sulfate)-GalNAc(6-O-sulfate) (D/iD) and GlcUA/IdoUA-GalNAc(4, 6-O-disulfate) (E/iE) decreased to 50 and 30%, respectively, in the developing cerebellum. These results suggest that the IdoUA-containing iA and iB units along with D/iD and E/iE units in the CS/DS hybrid play important roles in the formation of the cerebellar neural network during postnatal brain development.