Robson P, Stein P, Zhou B, Schultz R M, Baldwin H S
Division of Cardiology, Joseph Stokes Jr. Research Institute, Philadelphia, Pennsylvania 19104, USA.
Dev Biol. 2001 Jun 15;234(2):317-29. doi: 10.1006/dbio.2001.0274.
Platelet/Endothelial Cell Adhesion Molecule-1 (PECAM-1 or CD31) is thought to be a vascular-specific protein, but its function has not been clearly defined. Here, we demonstrate by using confocal immunofluorescence microscopy that PECAM-1 is first detected in the mouse blastocyst, which contains no vascular cells, and its expression is restricted to the pluripotent inner cell mass (ICM) cells. Expression is localized to cell-cell borders of the ICM and is detected at the very first signs of blastocoel formation. Consistent with these observations is that embryonic transcripts of PECAM-1 mRNA, as detected by RT-PCR, greatly increase during the morula-to-blastocyst transition and seven of the eight known alternatively spliced isoforms of PECAM-1 are expressed in the blastocyst. The synthesis of PECAM-1 is independent of compaction, cytokinesis, and DNA replication, as it is detected in embryos that are chronologically at the blastocyst stage following culture of 8-cell embryos in Ca2+-free medium, or medium containing cytochalasin D or aphidicolin, respectively. By the late blastocyst stage, PECAM-1 expression is restricted to the pluripotent epiblast, at which point it has a mutually exclusive expression pattern to that of type IV collagen, a basement membrane marker. The reduction in PECAM-1 transcripts in retinoic acid-induced differentiation of F9 teratocarcinoma cells, a model of epiblast-to-primitive endoderm differentiation, confirmed the epiblast-specific expression of PECAM-1. By the egg cylinder stage of development, at which point the epiblast is no longer pluripotent, PECAM-1 is not detected. This ICM-specific pattern of expression suggests a novel developmental role of PECAM-1 that is independent of its function in vascular ontogeny.
血小板/内皮细胞黏附分子-1(PECAM-1或CD31)被认为是一种血管特异性蛋白,但其功能尚未明确界定。在此,我们通过共聚焦免疫荧光显微镜证明,PECAM-1最早在不含血管细胞的小鼠囊胚中被检测到,其表达局限于多能内细胞团(ICM)细胞。表达定位于ICM的细胞-细胞边界,并在囊胚腔形成的最初迹象时被检测到。与这些观察结果一致的是,通过RT-PCR检测,PECAM-1 mRNA的胚胎转录本在桑椹胚向囊胚转变过程中大幅增加,并且PECAM-1的八种已知可变剪接异构体中的七种在囊胚中表达。PECAM-1的合成独立于紧密化、胞质分裂和DNA复制,因为在分别在无钙培养基、含有细胞松弛素D或阿非迪霉素的培养基中培养8细胞胚胎后,按时间顺序处于囊胚阶段的胚胎中检测到了它。到囊胚晚期,PECAM-1的表达局限于多能外胚层,此时它与基底膜标记物IV型胶原具有相互排斥的表达模式。视黄酸诱导F9畸胎瘤细胞分化(一种外胚层向原始内胚层分化的模型)过程中PECAM-1转录本的减少,证实了PECAM-1在外胚层中的特异性表达。到发育的卵圆柱期,此时外胚层不再具有多能性,未检测到PECAM-1。这种ICM特异性的表达模式表明PECAM-1具有一种新的发育作用,独立于其在血管发生中的功能。