Zhang Yi, Yeung Man-Nga, Liu Jun, Chau Chi-Ho, Chan Ying-Shing, Shum Daisy K Y
Department of Biochemistry, Faculty of Medicine, The University of Hong Kong, China.
J Comp Neurol. 2006 Jan 10;494(2):345-57. doi: 10.1002/cne.20811.
This work addresses the cellular localization of heparanase and its colocalization with syndecan-3, a transmembrane heparan sulfate proteoglycan in spinal cords of adult rats. Reverse transcriptase/polymerase chain reaction (RT-PCR) and in situ hybridization for the heparanase transcript revealed expression in neurons and white matter glia. This was confirmed by immunohistochemistry showing cytoplasmic localization of the heparanase protein. Double immunofluorescence for heparanase and syndecan-3 revealed colocalization of the proteins in cell bodies of neurons and oligodendrocytes, suggestive of constitutive expression in these cell types. In contrast, only subpopulations of astrocytes and NG2-expressing glia in the white matter expressed heparanase, and these did not show expression of syndecan-3. Cultures of astrocytes further evidenced upregulation of heparanase expression with TGF-beta(1) treatment, but no accompanying upregulation of syndecan-3 was detectable. These first findings of heparanase expression in the adult cord therefore provide the cellular basis for understanding functional interactions of heparanase and syndecan-3 in the normal neural network or otherwise in glial reactions to spinal cord injury.
本研究探讨了乙酰肝素酶在成年大鼠脊髓中的细胞定位及其与跨膜硫酸乙酰肝素蛋白聚糖Syndecan-3的共定位情况。通过逆转录酶/聚合酶链反应(RT-PCR)和乙酰肝素酶转录本的原位杂交,发现其在神经元和白质胶质细胞中表达。免疫组织化学显示乙酰肝素酶蛋白定位于细胞质,进一步证实了这一结果。乙酰肝素酶和Syndecan-3的双重免疫荧光显示,这两种蛋白在神经元和少突胶质细胞的细胞体中共定位,提示它们在这些细胞类型中组成性表达。相反,仅白质中的星形胶质细胞亚群和表达NG2的胶质细胞表达乙酰肝素酶,且这些细胞不表达Syndecan-3。星形胶质细胞培养进一步证明,转化生长因子-β1(TGF-β1)处理可上调乙酰肝素酶的表达,但未检测到Syndecan-3的伴随上调。因此,这些关于成年脊髓中乙酰肝素酶表达的初步发现为理解乙酰肝素酶和Syndecan-3在正常神经网络中的功能相互作用,或在脊髓损伤时胶质细胞反应中的功能相互作用提供了细胞基础。