Rappolee D A, Patel Y, Jacobson K
Department of Cell and Molecular Biology, Lurie Cancer Center, Northwestern University Medical School, Chicago, Illinois 60611, USA.
Mol Reprod Dev. 1998 Nov;51(3):254-64. doi: 10.1002/(SICI)1098-2795(199811)51:3<254::AID-MRD4>3.0.CO;2-O.
FGF receptor (FGFR) function is essential during peri-implantation mouse development. To understand which receptors are functioning, we tested for the expression of all four FGF receptors in peri-implantation blastocysts. By RT-PCR, FGFR-3 and FGFR-4 were detected at high levels, FGFR-2 at lower levels, and FGFR-1 was detected at background levels compared to control tissues. Because FGFR-3 and FGFR-4 were detected at the highest levels, we studied these in detail. Between 3.5 days after fertilization (E3.5) and E6.0, FGFR-4 mRNA was detected ubiquitously in the peri-implantation embryo, restricted to the inner cell mass (ICM) and its derivatives and primitive endoderm by E6.0, and was not detected at E6.5. FGFR-3 mRNA was detected ubiquitously in the peri-implantation embryo with a tendency towards extraembryonic cells. We tested blastocyst outgrowths, a model for implantation, for FGFR-3 and FGFR-4 protein. FGFR-3 protein was detected in all cells early during the outgrowth. Later, FGFR-3 was detected in the extraembryonic endoderm and trophoblast giant cells (TGC), but not in the ICM. FGFR-4 protein was detected in all cells of the implanting embryo, but was restricted to the ICM/primitive endoderm in later stage outgrowths. The distribution of the receptor proteins in the blastocyst outgrowths is similar to the distribution of the mRNA detected by in situ hybridization of sections of embryos. The data suggest roles for FGFR-3 and FGFR-4 in peri-implantation development.
成纤维细胞生长因子受体(FGFR)的功能在小鼠着床前期发育过程中至关重要。为了解哪些受体在发挥作用,我们检测了着床前期囊胚中所有四种FGFR的表达情况。通过逆转录聚合酶链反应(RT-PCR)发现,FGFR-3和FGFR-4的表达水平较高,FGFR-2的表达水平较低,而与对照组织相比,FGFR-1仅在背景水平被检测到。由于FGFR-3和FGFR-4的检测水平最高,我们对其进行了详细研究。在受精后3.5天(E3.5)至E6.0期间,FGFR-4 mRNA在着床前期胚胎中普遍存在,到E6.0时局限于内细胞团(ICM)及其衍生物和原始内胚层,在E6.5时未被检测到。FGFR-3 mRNA在着床前期胚胎中普遍存在,且有向胚外细胞富集的趋势。我们在囊胚体外生长模型(一种着床模型)中检测了FGFR-3和FGFR-4蛋白。在体外生长早期,所有细胞中均检测到FGFR-3蛋白。之后,在胚外内胚层和滋养层巨细胞(TGC)中检测到FGFR-3,但在ICM中未检测到。FGFR-4蛋白在着床胚胎的所有细胞中均有检测到,但在后期体外生长阶段局限于ICM/原始内胚层。受体蛋白在囊胚体外生长中的分布与通过胚胎切片原位杂交检测到的mRNA分布相似。这些数据表明FGFR-3和FGFR-4在着床前期发育中发挥作用。