LECTIN STAINING OF CULTURED CNS MICROGLIA
LECTIN STAINING OF CULTURED CNS MICROGLIA
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DOI：
10.1177/40.4.1372634
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发表时间：
1992-04-01
影响因子：
3.2
通讯作者：
YAO, JB
中科院分区：
文献类型：
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作者：
COLTON, CA;ABEL, C;YAO, JB
碳水化合物结合蛋白（称为凝集素）与大多数膜上的特定糖基结合。 我们使用荧光和光学显微镜来研究各种凝集素与新生大鼠或胎鼠大脑皮质培养的小胶质细胞膜的相互作用。 小胶质细胞被 GS-1、RCA、WGA 和 ConA 强烈染色，而被 DBA、UEA、BPA 和 SBA 轻微染色。 GS-2、MPA 或 PNA 未见染色。 染色对混合神经胶质培养物中的小胶质细胞具有特异性，并且具有剂量依赖性。 此外，使用特定糖的竞争性抑制可以减少或阻断小胶质细胞凝集素的结合。 用二甲基亚砜 (DMSO)、白细胞介素-1 (IL-1)、干扰素 (IFN) 或脂多糖 (LPS) 等药物处理小胶质细胞并不能消除凝集素染色，尽管染色程度发生了改变。 小胶质细胞的阳性染色还与至少一种凝集素（即 ConA）的功能变化相关。 ConA 存在时，小胶质细胞产生的超氧阴离子增加。 总体而言，凝集素 GS-1、RCA、WGA 和 ConA 的结合可用作神经胶质培养物中小胶质细胞的识别工具，但染色强度根据其功能状态而变化。
Carbohydrate binding proteins, known as lectins, bind to specific sugar groups on most membranes. We used fluorescent and light microscopy to study the interaction of various lectins with the membranes of microglia cultured from neonatal rat or fetal mouse cerebral cortices. Microglia stained intensely with GS-1, RCA, WGA, and ConA and slightly with DBA, UEA, BPA, and SBA. No staining was seen with GS-2, MPA, or PNA. Staining was specific for microglia in the mixed glial cultures and was dose dependent. In addition, microglial lectin binding could be reduced or blocked by competitive inhibition using specific sugars. Treatment of the microglia with agents such as dimethylsulfoxide (DMSO), interleukin-1 (IL-1), interferon (IFN), or lipopolysaccharide (LPS) did not eliminate lectin staining, although the degree of staining was altered. Positive staining of the microglia was also associated with a functional change for at least one lectin, i.e., ConA. Superoxide anion production by microglia was increased in the presence of ConA. Overall, binding of the lectins GS-1, RCA, WGA, and ConA can be used as an identifying tool for microglia in glial cultures, but intensity of staining varies depending on their functional state.