Hennig A, Krueger R, Mangoura D, Schwartz N B
Department of Pediatrics, University of Chicago, IL 60637.
Cell Mol Biol (Noisy-le-grand). 1992 Aug-Sep;38(5-6):585-93.
Proteoglycans of developing chick brain were distinguished on the basis of reactivity with four well characterized antibody reagents (S103L, to the CS-rich domain; HNK-1, to 6-sulfated glucuronic acid; 1-C-3, to the HABr region and 5-D-4, to KS chains). One chondroitin sulfate proteoglycan reacted exclusively with S103L and 1-C-3 and not with the other two antibodies, hence is designated the S103L reactive brain CSPG. The other proteoglycan reacted exclusively with HNK-1 and 5-D-4 and not with S103L and 1-C-3, hence it is designated the HNK-1 reactive brain CSPG. In addition to these immunological distinctions, the S103L and HNK-1 CSPGs exhibited significant biochemical differences at both the protein and carbohydrate levels. Most interestingly, both CSPGs were found in all regions of the brain, and were expressed in a developmentally regulated pattern. The S103L CSPG was not detectable prior to embryonic day 7, increased to a maximum at day 13-15 and declined by day 20 in most brain regions examined. In contrast, the HNK-1 CSPG was present as early as embryonic day 4 and remained constant through hatching. Neuronal cultures established from embryonic day 6 (E6) cerebral hemispheres represent an in vitro paradigm that mimics in vivo neuronal development and differentiation. In this culture system we found that the expression of the S103L and HNK-1 CSPG followed a pattern similar to that observed in developing brain and further, that neurons are probably the sole source of S103L CSPG in cerebral cortex during neuroembryogenesis.
根据与四种特征明确的抗体试剂(针对富含硫酸软骨素(CS)结构域的S103L、针对6 - 硫酸化葡萄糖醛酸的HNK - 1、针对HABr区域的1 - C - 3以及针对硫酸角质素(KS)链的5 - D - 4)的反应性,对发育中的鸡脑蛋白聚糖进行了区分。一种硫酸软骨素蛋白聚糖仅与S103L和1 - C - 3反应,而不与其他两种抗体反应,因此被命名为S103L反应性脑硫酸软骨素蛋白聚糖(CSPG)。另一种蛋白聚糖仅与HNK - 1和5 - D - 4反应,而不与S103L和1 - C - 3反应,因此它被命名为HNK - 1反应性脑CSPG。除了这些免疫学差异外,S103L和HNK - 1 CSPG在蛋白质和碳水化合物水平上还表现出显著的生化差异。最有趣的是,两种CSPG在脑的所有区域均有发现,并以发育调控的模式表达。在大多数检测的脑区域中,S103L CSPG在胚胎第7天之前无法检测到,在第13 - 15天增加到最大值,并在第20天下降。相比之下,HNK - 1 CSPG早在胚胎第4天就已存在,并在孵化过程中保持恒定。从胚胎第6天(E6)大脑半球建立的神经元培养物代表了一种模拟体内神经元发育和分化的体外模型。在这个培养系统中,我们发现S103L和HNK - 1 CSPG的表达遵循与发育中的脑相似的模式,并且进一步发现,在神经胚胎发生过程中,神经元可能是大脑皮层中S103L CSPG的唯一来源。