Cytochemical identification of turbot myeloperoxidase-positive granulocytes by potassium iodide and oxidized pyronine Y staining
Cytochemical identification of turbot myeloperoxidase-positive granulocytes by potassium iodide and oxidized pyronine Y staining
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碘化钾和氧化派洛宁 Y 染色对大菱鲆髓过氧化物酶阳性粒细胞的细胞化学鉴定
DOI：
10.1016/j.tice.2017.10.008
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发表时间：
2017-12-01
期刊：
影响因子：
2.6
通讯作者：
Sun, Li
中科院分区：
文献类型：
--
作者：
Chi, Heng;Wen, Li-lian;Sun, Li
细胞形态学和细胞化学染色是鉴定细胞类型的重要方法，特别是在通常缺乏生物工具如特异性抗体的鱼类中。髓过氧化物酶（MPO）通常用作组织中中性粒细胞蓄积的细胞内标志物和血浆中中性粒细胞活性的标志物。在这项研究中，我们报告了一种碘化钾和氧化派罗宁Y（KI PyY）染色方法，用于快速，高灵敏度检测大菱鲆血液，腹膜和组织中的MPO阳性细胞。MPO阳性细胞（主要代表中性粒细胞）被染成棕色，并与其他细胞（如淋巴细胞、单核细胞和巨噬细胞）明显区分，后者被染成粉红色。经细菌刺激后，中性粒细胞在外周血白细胞和腹膜中所占比例分别为27.49%和38.05%。肾粒细胞中含有丰富的MPO阳性细胞，可能是MPO低表达的幼稚中性粒细胞。值得注意的是，在肾、脾和肠的组织切片中检测到MPO阳性细胞，其分布特征特异于每个组织。然而，在染色的组织切片中，细胞形态不明显。结果表明，KI-PyY染色法灵敏度高，适用于不同类型的样品，可用于鱼类不同区室中性粒细胞的研究。
Cytomorphological and cytochemical staining are important methods for the identification of cell types, in particular in fish which often lack biological tools such as specific antibodies. Myeloperoxidase (MPO) is usually used as an intracellular marker of neutrophil accumulation in tissues and a marker of neutrophil activity in plasma. In this study, we reported a potassium iodide and oxidized pyronine Y (KI-PyY) staining method for rapid and highly sensitive detection of MPO-positive cells in turbot blood, peritoneum, and tissues. MPO-positive cells, which mostly represented neutrophils, were stained brown and clearly distinguished from other cells, such as lymphocytes, monocytes, and macrophages, which were stained pink. Following bacterial stimulation, the proportions of neutrophils were 27.49% and 38.05% in peripheral blood leukocytes and peritoneum, respectively, judging by the stained MPO. Kidney granulocytes contained abundant MPO-positive cells which were probably immature neutrophils with low expression of MPO. It is noteworthy that MPO-positive cells were detected in the tissue sections of kidney, spleen, and gut, with distribution profiles specific to each tissue. However, the cell morphology was not distinct in the stained tissue sections. These results indicate that the KI-PyY staining method is highly sensitive, applicable to different types of samples, and will be useful for the study of neutrophils in different compartments of fish.