Gründker C, Kirchner C
Department of Biology, Philipps University, Marburg, Germany.
Anat Embryol (Berl). 1996 Aug;194(2):169-75. doi: 10.1007/BF00195010.
Fibroblast growth factor-2 (FGF-2) induces gastrulation of rabbit blastocysts in vitro and is present in the uterine secretion at day 6 after mating. The following study was made in order to show if changes in the uterine FGF-2 concentration or in the FGF receptor concentration of the embryonic tissues point to a regulation of this event. By the use of the ELISA technique and immunohistochemistry, FGF-2 concentration was determined in the endometrial tissue, uterine secretion and blastocyst between day 4 and day 8 of pregnancy, in the uterine secretion after induction of pseudopregnancy, in day 6 blastocysts after in vitro culture, and FGF immunoreactivity was localized in the endometrial tissue. FGF receptor-1 (FGFR-1) concentration was examined correspondingly in the blastocyst. Cross-linking experiments using 125I-FGF-2 were done to identify binding proteins in the blastocyst. In the uterine secretion, FGF-2 was constantly high up to day 6.5 but showed an increase thereafter. Similar values in pseudopregnant uterine secretions indicated that the growth factor was of uterine origin. It was probably synthesized by the uterine epithelium as shown by immunohistochemistry. Under culturing conditions, the blastocyst produced small amounts of FGF-2. In the blastocyst, FGFR-1 as well as binding of 125I-FGF-2 showed a dramatic increase from day 6.0 to day 6.5, coinciding with the onset of gastrulation. Receptor antigenicity was located in the embryonic disc at day 6.5 and day 7.0. Two binding proteins of about 200 and 130 kDa were found by cross-linking. The results indicate that a regulation of growth factor influence on embryonic differentiation is more probable via expression of the embryonic receptor than via differential release of the uterine growth factor.
成纤维细胞生长因子-2(FGF-2)可在体外诱导兔胚泡发生原肠胚形成,且在交配后第6天存在于子宫分泌物中。进行以下研究是为了表明子宫FGF-2浓度或胚胎组织中FGF受体浓度的变化是否表明该事件存在调控。通过使用酶联免疫吸附测定(ELISA)技术和免疫组织化学方法,测定了妊娠第4天至第8天子宫内膜组织、子宫分泌物和胚泡中的FGF-2浓度,假孕诱导后子宫分泌物中的FGF-2浓度,体外培养后第6天胚泡中的FGF-2浓度,并将FGF免疫反应性定位在子宫内膜组织中。相应地检测了胚泡中FGF受体-1(FGFR-1)的浓度。使用125I-FGF-2进行交联实验以鉴定胚泡中的结合蛋白。在子宫分泌物中,FGF-2在第6.5天之前一直保持较高水平,但此后有所增加。假孕子宫分泌物中的相似值表明该生长因子源自子宫。免疫组织化学显示它可能由子宫上皮合成。在培养条件下,胚泡产生少量FGF-2。在胚泡中,FGFR-1以及125I-FGF-2的结合从第6.0天到第6.5天显著增加,这与原肠胚形成的开始相吻合。在第6.5天和第7.0天,受体抗原性位于胚盘。通过交联发现了两种分子量约为200 kDa和130 kDa的结合蛋白。结果表明,生长因子对胚胎分化的影响更可能是通过胚胎受体的表达来调控,而不是通过子宫生长因子的差异释放来调控。