Parmley R T, Takagi M, Spicer S S, Thrasher A, Denys F R
Am J Anat. 1982 Sep;165(1):53-67. doi: 10.1002/aja.1001650106.
Complex carbohydrates in eosinophils from human, rabbit, and rat marrow were identified and localized by cytochemical and radioautographic methods. The high iron diamine-thiocarbohydrazide-silver proteinate (HID-TCH-SP), low iron diamine (LID)-TCH-SP, and periodate (PA)-TCH-SP methods were used for the localization of sulfate, sulfate and carboxyl, and vicinal glycol- containing complex carbohydrates, respectively. Golgi vesicles and small precursor granules (0.2-0.4 micrometer in diameter) demonstrated strong HID-TCH-SP staining and labeled intensely after a 10-minute pulse with 35SO4(2). Crystalloid-free or immature specific granules (0.5-0.9 micrometers in diameter) labeled heavily after a 60-minute incubation and 60-minute chase with 35SO4(2). Immature granules were graded according to their HID-TCH-SP staining. Type I granules demonstrated strong rim staining and similar or somewhat less central staining, whereas type 2 granules only demonstrated rim staining, and type 3 granules lacked staining. Fully mature crystalloid-containing granules lacked staining. LID-TCH-SP similarly stained the HID-positive sulfated material in cytoplasmic granules. PA-TCH-SP stained some Golgi vesicles and diffusely stained all precursor granules and type 1 granules. Weaker staining was observed in type 2 granules and staining was very weak or absent in type 3 and crystalloid-containing granules. In early eosinophils, tubulovesicular structures (TVS) were observed rosetting and contacting precursor and type 1 granules. These TVS contained material with strong PA-TCH-SP staining but lacked HID-TCH-SP or LID-TCH-SP-reactive acidic glycoconjugates. Flattened Golgi saccules or early eosinophils stained weakly or not at all with the PA-TCH-SP method. Small granules and TVS in late (bilobed) eosinophils displayed PA-TCH-SP reactivity and lacked HID-TCH-SP staining but differed from TVS in early eosinophils in that they were not associated as rosettes with specific granules. These results indicate that sulfated and vicinal glycol-containing complex carbohydrates are differently disturbed in immature specific granules of eosinophils and presumably become masked to staining as the granule matures.
采用细胞化学和放射自显影方法，对人、兔和大鼠骨髓嗜酸性粒细胞中的复合碳水化合物进行了鉴定和定位。高铁二胺-硫代碳酰肼-蛋白银（HID-TCH-SP）法、低铁二胺（LID）-TCH-SP法和高碘酸盐（PA）-TCH-SP法分别用于定位硫酸盐、硫酸盐和羧基以及含邻位二醇的复合碳水化合物。高尔基体小泡和小前体颗粒（直径0.2 - 0.4微米）显示出强烈的HID-TCH-SP染色，在用35SO4(2)脉冲10分钟后标记强烈。无晶体或未成熟的特异性颗粒（直径0.5 - 0.9微米）在与35SO4(2)孵育60分钟并追踪60分钟后标记强烈。未成熟颗粒根据其HID-TCH-SP染色进行分级。I型颗粒显示出强烈的边缘染色和相似或稍弱的中央染色，而2型颗粒仅显示边缘染色，3型颗粒缺乏染色。完全成熟的含晶体颗粒缺乏染色。LID-TCH-SP同样对细胞质颗粒中HID阳性的硫酸化物质进行染色。PA-TCH-SP对一些高尔基体小泡进行染色，并对所有前体颗粒和1型颗粒进行弥漫性染色。在2型颗粒中观察到较弱的染色，在3型和含晶体颗粒中染色非常弱或无染色。在早期嗜酸性粒细胞中，观察到管状泡状结构（TVS）呈玫瑰花结状并与前体和1型颗粒接触。这些TVS含有具有强烈PA-TCH-SP染色的物质，但缺乏HID-TCH-SP或LID-TCH-SP反应性酸性糖缀合物。扁平的高尔基体囊泡或早期嗜酸性粒细胞用PA-TCH-SP法染色较弱或根本不染色。晚期（双叶）嗜酸性粒细胞中的小颗粒和TVS显示出PA-TCH-SP反应性，缺乏HID-TCH-SP染色，但与早期嗜酸性粒细胞中的TVS不同，它们不作为玫瑰花结与特异性颗粒相关联。这些结果表明，硫酸盐和含邻位二醇的复合碳水化合物在嗜酸性粒细胞未成熟的特异性颗粒中受到不同的干扰，并且随着颗粒成熟，可能会对染色产生掩盖作用。
