Constantinescu Dan, Gray Heather L, Sammak Paul J, Schatten Gerald P, Csoka Antonei B
Department of Cell Biology and Physiology, University of Pittsburgh School of Medicine, Pittsburgh Development Center of Magee-Womens Research Institute, 204 Craft Ave, Pittsburgh, Pennsylvania 15213, USA.
Stem Cells. 2006 Jan;24(1):177-85. doi: 10.1634/stemcells.2004-0159. Epub 2005 Sep 22.
Nuclear lamins comprise the nuclear lamina, a scaffold-like structure that lines the inner nuclear membrane. B-type lamins are present in almost all cell types, but A-type lamins are expressed predominantly in differentiated cells, suggesting a role in maintenance of the differentiated state. Previous studies have shown that lamin A/C is not expressed during mouse development before day 9, nor in undifferentiated mouse embryonic carcinoma cells. To further investigate the role of lamins in cell phenotype maintenance and differentiation, we examined lamin expression in undifferentiated mouse and human embryonic stem (ES) cells. Wide-field and confocal immunofluorescence microscopy and semiquantitative reverse transcription-polymerase chain reaction analysis revealed that undifferentiated mouse and human ES cells express lamins B1 and B2 but not lamin A/C. Mouse ES cells display high levels of lamins B1 and B2 localized both at the nuclear periphery and throughout the nucleoplasm, but in human ES cells, B1 and B2 expression is dimmer and localized primarily at the nuclear periphery. Lamin A/C expression is activated during human ES cell differentiation before downregulation of the pluripotency marker Oct-3/4 but not before the downregulation of the pluripotency markers Tra-1-60, Tra-1-81, and SSEA-4. Our results identify the absence of A-type lamin expression as a novel marker for undifferentiated ES cells and further support a role for nuclear lamins in cell maintenance and differentiation.
核纤层蛋白构成核纤层,这是一种位于内核膜内侧的支架样结构。B型核纤层蛋白存在于几乎所有细胞类型中,但A型核纤层蛋白主要在分化细胞中表达,这表明其在维持分化状态中发挥作用。先前的研究表明,在小鼠发育的第9天之前,以及在未分化的小鼠胚胎癌细胞中,lamin A/C均不表达。为了进一步研究核纤层蛋白在细胞表型维持和分化中的作用,我们检测了未分化的小鼠和人类胚胎干细胞(ES细胞)中的核纤层蛋白表达。宽视野和共聚焦免疫荧光显微镜以及半定量逆转录-聚合酶链反应分析显示,未分化的小鼠和人类ES细胞表达核纤层蛋白B1和B2,但不表达lamin A/C。小鼠ES细胞中核纤层蛋白B1和B2水平较高,定位于核周和整个核质中,但在人类ES细胞中,B1和B2的表达较暗淡,主要定位于核周。在人类ES细胞分化过程中,lamin A/C的表达在多能性标记物Oct-3/4下调之前被激活,但在多能性标记物Tra-1-60、Tra-1-81和SSEA-4下调之前未被激活。我们的结果确定缺乏A型核纤层蛋白表达是未分化ES细胞的一种新标记,并进一步支持核纤层蛋白在细胞维持和分化中的作用。