晶状体上皮细胞增殖活性的研究及组织型干细胞的鉴定。

A study of the proliferating activity in lens epithelium and the identification of tissue-type stem cells.

作者信息

Yamamoto Naoki, Majima Kiyoyuki, Marunouchi Tohru

机构信息

Laboratory of Molecular Biology and Histochemistry, Fujita Health University Joint Research Laboratory, Aichi, Japan.

出版信息

Med Mol Morphol. 2008 Jun;41(2):83-91. doi: 10.1007/s00795-008-0395-x. Epub 2008 Jul 1.

Abstract

Using a newly established fixation method and immunohistochemical methods, we precisely described the regions of cells stained with various antibodies relating to cell proliferation; this method enabled us to make cellular-level diagrams of the epithelium in which the position of every lens epithelial cell (LEC) was determined in reference to the cell located at the top of the bow area. The proliferating activity of LECs of 4-week-old (4W) mice was examined either by labeling with 5-bromodeoxyuridine (BrdU) in vivo or by measuring the amount of mRNA prepared from LECs, which had been separated into the posterior part, containing the germinative zone, and the anterior part and then cultured. The epithelial region stained with antibody for proliferating cell nuclear antigen (PCNA) and cyclin D1 remained relatively constant during the study period, although the positive region was reduced a little from embryonic day 18 (E18) to 12W. This region at 4W overlapped well with the DNA synthesizing region. Therefore, we reasoned that this region would correspond to the germinative zone of the adult mouse. Considering together with results of the reactivation pattern of genes, we considered that the location of tissue-type stem cells in lens epithelium (LE) as immediately anterior to the germinative zone.

摘要

我们使用一种新建立的固定方法和免疫组织化学方法,精确描述了用与细胞增殖相关的各种抗体染色的细胞区域;这种方法使我们能够绘制上皮细胞的细胞水平图,其中每个晶状体上皮细胞(LEC)的位置是参照位于弓状区域顶部的细胞确定的。通过在体内用5-溴脱氧尿苷(BrdU)标记或通过测量从LEC制备的mRNA量来检测4周龄(4W)小鼠LEC的增殖活性,这些LEC已被分离为包含生发区的后部和前部,然后进行培养。在用增殖细胞核抗原(PCNA)和细胞周期蛋白D1抗体染色的上皮区域在研究期间保持相对恒定,尽管从胚胎第18天(E18)到12W阳性区域略有减少。4W时的该区域与DNA合成区域很好地重叠。因此,我们推断该区域将对应于成年小鼠的生发区。结合基因重新激活模式的结果,我们认为晶状体上皮(LE)中组织型干细胞的位置紧邻生发区前方。

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