Localization of chondroitin sulfate proteoglycan versican in adult brain with special reference to large projection neurons
Localization of chondroitin sulfate proteoglycan versican in adult brain with special reference to large projection neurons
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DOI：
10.1007/s00441-008-0698-1
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发表时间：
2008-11-01
影响因子：
3.6
通讯作者：
Wanaka, Akio
中科院分区：
文献类型：
--
作者：
Horii-Hayashi, Noriko;Okuda, Hiroaki;Wanaka, Akio
Versican是一种属于lectican家族的硫酸软骨素蛋白聚糖。Versican具有两个糖胺聚糖附着区，称为GAG α和GAG β结构域，其均受选择性剪接调节并产生四种蛋白质同种型。我们已经研究了多能蛋白聚糖的表达和定位在发展和成人大脑中使用抗多能蛋白聚糖GAG α和GAG β抗体。Western分析显示，GAG α-反应亚型在成人脑中占主导地位。免疫组化研究表明，GAG α免疫反应性可检测到从新生儿期到成年，而GAG β免疫反应性完全消失，在出生3周内。在成人脑中，在白色物质区域观察到GAG α免疫反应性，并且还定位于灰质中，包括皮质和海马锥体神经元和小脑浦肯野细胞的胞体和树突。相比之下，GAG α免疫反应性不局限于小清蛋白阳性中间神经元和小脑星状细胞。此外，GAG α免疫反应性不与神经元周网标记物（如紫藤凝集素凝集素和磷酸蛋白聚糖）共定位。因此，多功能蛋白聚糖定位于大的投射神经元，而不是小的中间神经元。为了证实多功能蛋白聚糖与神经元的结合机制，在多功能蛋白聚糖的免疫标记之前，从脑切片中酶促除去透明质酸和硫酸软骨素。这些处理对多功能蛋白聚糖的标记模式没有影响，这表明其他多功能蛋白聚糖相互作用分子参与了多功能蛋白聚糖与神经元的结合。
Versican is a chondroitin sulfate proteoglycan belonging to the lectican family. Versican has two glycosaminoglycan attachment regions, named the GAG alpha and GAG beta domains, which are both regulated by alternative splicing and yield four protein isoforms. We have investigated the expression and localization of versican in the developing and adult brain by using anti-versican GAG alpha and GAG beta antibodies. Western analysis revealed that GAG alpha-reactive isoform was dominant in the adult brain. Immunohistochemical study demonstrated that GAG alpha immunoreactivity was detectable from neonatal periods to adulthood, whereas GAG beta immunoreactivity completely disappeared within 3 weeks of birth. In the adult brain, GAG alpha immunoreactivity was seen in the white matter regions and was also localized in the gray matter including somata and dendrites of cortical and hippocampal pyramidal neurons and cerebellar Purkinje cells. In contrast, GAG alpha immunoreactivity was not localized on parvalbumin-positive interneurons and cerebellar stellate cells. Furthermore, GAG alpha immunoreactivity was not co-localized with perineuronal net markers such as Wisteria floribunda agglutinin lectin and phosphacan. Thus, versican was localized on large projection neurons rather than small interneurons. To confirm the binding mechanism of versican to neurons, hyaluronan and chondroitin sulfates were enzymatically removed from brain sections before the immunolabeling of versican. These treatments had no effect on the labeling pattern of versican, suggesting that other versican-interactive molecules are involved in the binding of versican to neurons.