Leung Anskar Y H, Leung Joseph C K, Chan Loretta Y Y, Ma Edmond S K, Kwan Tommy T F, Lai K N, Meng A, Liang Raymond
Department of Medicine, Queen Mary Hospital, University of Hong Kong, Pok Fu Lam Road, Hong Kong, Hong Kong.
Histochem Cell Biol. 2005 Aug;124(2):105-11. doi: 10.1007/s00418-005-0003-2. Epub 2005 Sep 29.
We investigated the expression of proliferative cell nuclear antigen (PCNA) in zebrafish to delineate the proliferative hematopoietic component during adult and embryonic hematopoiesis. Immunostaining for PCNA and enhanced green fluorescence protein (eGFP) was performed in wild-type and fli1-eGFP (endothelial marker) and gata1-eGFP (erythroid cell marker) transgenic fish. Expression of PCNA mRNA was examined in wild-type and chordin morphant embryos. In adult zebrafish kidney, the renal tubules are surrounded by endothelial cells and it is separated into hematopoietic and excretory compartments. PCNA was expressed in hematopoietic progenitor cells but not in mature neutrophils, eosinophils or erythroid cells. Some PCNA+ cells are scattered in the hematopoietic compartment of the kidney while others are closely associated with renal tubular cells. PCNA was also expressed in spermatogonial stem cells and intestine crypts, consistent with its role in cell proliferation and DNA synthesis. In embryos, PCNA is expressed in the brain, spinal cord and intermediate cell mass (ICM) at 24 h-post fertilization. In chordin morphants, PCNA is significantly upregulated in the expanded ICM. Therefore, PCNA can be used to mark cell proliferation in zebrafish hematopoietic tissues and to identify a population of progenitor cells whose significance would have to be further investigated.
我们研究了增殖细胞核抗原(PCNA)在斑马鱼中的表达,以描绘成年和胚胎造血过程中增殖性造血成分。在野生型、fli1-eGFP(内皮细胞标记物)和gata1-eGFP(红细胞标记物)转基因鱼中进行了PCNA和增强型绿色荧光蛋白(eGFP)的免疫染色。检测了野生型和脊索蛋白 morphant 胚胎中PCNA mRNA的表达。在成年斑马鱼肾脏中,肾小管被内皮细胞包围,且肾脏被分为造血和排泄区室。PCNA在造血祖细胞中表达,但在成熟的中性粒细胞、嗜酸性粒细胞或红细胞中不表达。一些PCNA+细胞散在于肾脏的造血区室中,而其他细胞则与肾小管细胞紧密相关。PCNA也在精原干细胞和肠隐窝中表达,这与其在细胞增殖和DNA合成中的作用一致。在胚胎中,受精后24小时,PCNA在脑、脊髓和中间细胞团(ICM)中表达。在脊索蛋白morphant中,PCNA在扩大的ICM中显著上调。因此,PCNA可用于标记斑马鱼造血组织中的细胞增殖,并鉴定一群祖细胞,其重要性有待进一步研究。