Chi Heng, Wen Li-Lian, Sui Zhi-Hai, Sun Qing-Lei, Sun Li
Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China.
Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China; University of Chinese Academy of Sciences, Beijing, China.
Tissue Cell. 2017 Dec;49(6):751-755. doi: 10.1016/j.tice.2017.10.008. Epub 2017 Nov 3.
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.
细胞形态学和细胞化学染色是鉴定细胞类型的重要方法，特别是在鱼类中，因为鱼类通常缺乏诸如特异性抗体等生物学工具。髓过氧化物酶（MPO）通常用作组织中嗜中性粒细胞聚集的细胞内标志物以及血浆中嗜中性粒细胞活性的标志物。在本研究中，我们报道了一种碘化钾和氧化吡罗红Y（KI-PyY）染色方法，用于快速、高度灵敏地检测大菱鲆血液、腹膜和组织中的MPO阳性细胞。MPO阳性细胞大多代表嗜中性粒细胞，被染成棕色，与其他细胞如淋巴细胞、单核细胞和巨噬细胞（被染成粉红色）明显区分开来。经细菌刺激后，根据MPO染色判断，外周血白细胞和腹膜中嗜中性粒细胞的比例分别为27.49%和38.05%。肾粒细胞含有丰富的MPO阳性细胞，这些细胞可能是MPO表达较低的未成熟嗜中性粒细胞。值得注意的是，在肾脏、脾脏和肠道的组织切片中检测到了MPO阳性细胞，且各组织具有特定的分布特征。然而，在染色的组织切片中细胞形态并不明显。这些结果表明，KI-PyY染色方法高度灵敏，适用于不同类型的样本，将有助于研究鱼类不同部位的嗜中性粒细胞。
