Meijer Annemarie H, van der Sar Astrid M, Cunha Cristiana, Lamers Gerda E M, Laplante Mary A, Kikuta Hiroshi, Bitter Wilbert, Becker Thomas S, Spaink Herman P
Institute of Biology, Leiden University, Wassenaarseweg 64, 2333 AL Leiden, The Netherlands.
Dev Comp Immunol. 2008;32(1):36-49. doi: 10.1016/j.dci.2007.04.003. Epub 2007 May 22.
By enhancer trap screening we identified a transgenic zebrafish line showing leukocyte-specific YFP expression during late embryo and early larval development. Its enhancer detection insertion was mapped near a novel member of the myc proto-oncogene family, encoding transcription factors known to be important for regulating human myelopoiesis. Characterization of the zebrafish myc family showed that only this particular myc gene is strongly expressed in leukocytes. To identify the myc/YFP-expressing cell type, we re-examined specificity of described myeloid markers by multiplex fluorescent in situ hybridization, showing that lcp1 can be considered as a general leukocyte marker, csf1r as a macrophage-specific marker, and mpx and lyz as neutrophil-specific markers. Subsequent colocalization analysis defined the YFP-positive cells as a subset of the neutrophil population. Using real-time confocal imaging we demonstrate that these cells migrate to sites of inflammation and are involved in innate immune responses towards infections, including Mycobacterium marinum-induced granuloma formation.
通过增强子捕获筛选,我们鉴定出一条转基因斑马鱼品系,其在胚胎晚期和幼体早期发育过程中表现出白细胞特异性的黄色荧光蛋白(YFP)表达。其增强子检测插入位点被定位在原癌基因myc家族的一个新成员附近,该家族编码的转录因子已知对调节人类骨髓生成很重要。斑马鱼myc家族的特征表明,只有这个特定的myc基因在白细胞中强烈表达。为了鉴定表达myc/YFP的细胞类型,我们通过多重荧光原位杂交重新检查了所述髓系标志物的特异性,结果表明lcp1可被视为一般白细胞标志物,csf1r为巨噬细胞特异性标志物,mpx和lyz为中性粒细胞特异性标志物。随后的共定位分析将YFP阳性细胞定义为中性粒细胞群体的一个子集。使用实时共聚焦成像,我们证明这些细胞迁移到炎症部位,并参与对感染的固有免疫反应,包括由海分枝杆菌诱导的肉芽肿形成。