Phosphotyrosine antibodies specifically label ameboid microglia in vitro and ramified microglia in vivo.
Phosphotyrosine antibodies specifically label ameboid microglia in vitro and ramified microglia in vivo.
复制标题
磷酸酪氨酸抗体在体外特异性标记阿米巴样小胶质细胞，在体内特异性标记分支小胶质细胞。
作者：
Tillotson,ML;Wood,JG
使用亲和纯化的磷酸酪氨酸多克隆抗体（Wang：分子和细胞生物学5：3640-3643，1985），我们之前已经证明磷酸酪氨酸免疫反应性仅限于成年大鼠大脑中的多极 GFAP 阴性神经胶质细胞（Tillotson 和 Wood：比较神经学杂志282：133-141，1989）和视网膜（Tillotson 和 Wood：细胞生物学杂志 107：724a，1988）。在这项研究中，我们证明磷酸酪氨酸免疫反应细胞是小胶质细胞。磷酸酪氨酸免疫反应细胞和小胶质细胞之间的许多形态和超微结构相似性支持了这一结论。此外，磷酸酪氨酸与 Bandeiraea simplifolia-1 凝集素的小胶质细胞特异性 B4 异凝集素共定位。磷酸酪氨酸抗体还可对新生大鼠脑原代培养物中的阿米巴样小胶质细胞进行染色。此外，体外 7 天后，小胶质细胞是培养物中唯一的磷酸酪氨酸免疫反应元件。这种体外染色的时间模式模拟了原位磷酸酪氨酸免疫反应性的发育进程，其中多种结构在出生后神经发育过程中被染色（Tillotson and Wood:Journal of Comparison Neurology282:133–141, 1989），但只有成熟大脑中的小胶质细胞被染色。讨论了可能以小胶质细胞特异性方式表达的含磷酸酪氨酸的蛋白质的重要性。
Using an affinity‐purified, polyclonal antibody to phosphotyrosine (Wang:Molecular and Cellular Biology5:3640–3643, 1985) we have previously demonstrated that phosphotyrosine immunoreactivity is restricted to a population of multipolar GFAP‐negative neuroglia in adult rat brain (Tillotson and Wood:Journal of Comparative Neurology282:133–141, 1989) and retina (Tillotson and Wood:Journal of Cell Biology107:724a, 1988). In this study, we show that the phosphotyrosine‐immunoreactive cells are microglia. This conclusion is supported by numerous morphological and ultrastructural similarities between the phosphotyrosine‐immunoreactive cells and microglia. Furthermore, phosphotyrosine co‐localizes with the microglial‐specific B4isolectin ofBandeiraea simplifolia‐1 lectin. Phosphotyrosine antibodies also stain ameboid microglia in primary cultures of neonatal rat brain. In addition, after 7 days in vitro, microglia are the only phosphotyrosine‐immunoreactive element in the cultures. This temporal pattern of staining in vitro mimics the developmental progression of phosphotyrosine immunoreactivity in situ, in which a variety of structures stain during postnatal neural development (Tillotson and Wood:Journal of Comparative Neurology282:133–141, 1989), but only microglia stain in mature brain. The significance of phosphotyrosine‐containing proteins potentially expressed in a microglial‐specific manner is discussed.