Gulbis B, Jauniaux E, Cotton F, Stordeur P
Department of Clinical Chemistry, Hôpital Erasme, Université Libre de Bruxelles, Brussels, Belgium.
Mol Hum Reprod. 1998 Sep;4(9):857-62. doi: 10.1093/molehr/4.9.857.
The potential absorptive role of the yolk sac membrane was evaluated by examining protein and enzyme patterns in embryonic fluids and by comparing the synthetic capacity of the secondary yolk sac, fetal liver and placenta for human chorionic gonadotrophin (HCG) and alpha-fetoprotein (alphaFP). In yolk sac fluid samples, protein electrophoresis showed two main electrophoretic bands with mobilities comparable to those of albumin and interalbumin-alpha1-globulin, and immunoblotting revealed the presence of albumin, alphaFP, alpha1-antitrypsin, alpha2-macroglobulin, transferrin, complement factors 3 and 4 and immunoglobulin G. In coelomic fluid, similar results were obtained, except for the absence of alpha2-macroglobulin and the presence of ceruloplasmin and IgA. After electrophoresis and immunoblotting with specific antibodies, beta-HCG was detected in all placental homogenates and culture media but was not revealed in any of the corresponding yolk sac tissue samples. Reverse transcription-polymerase chain reaction (RT-PCR) showed that all placental samples express beta-HCG mRNA whereas all yolk sac and liver samples express alphaFP mRNA. These findings suggest that the yolk sac membrane is an important zone of transfer between the extra-embryonic and embryonic compartments and may also help to further develop therapeutic protocols making use of fetal somatic gene therapy by injecting transduced cells into the exocoelomic cavity.
通过检测胚胎液中的蛋白质和酶模式,以及比较次级卵黄囊、胎儿肝脏和胎盘合成人绒毛膜促性腺激素(HCG)和甲胎蛋白(αFP)的能力,评估卵黄囊膜的潜在吸收作用。在卵黄囊液样本中,蛋白质电泳显示出两条主要电泳带,其迁移率与白蛋白和白蛋白间α1球蛋白的迁移率相当,免疫印迹显示存在白蛋白、αFP、α1抗胰蛋白酶、α2巨球蛋白、转铁蛋白、补体因子3和4以及免疫球蛋白G。在体腔液中也获得了类似结果,只是不存在α2巨球蛋白,而存在铜蓝蛋白和IgA。用特异性抗体进行电泳和免疫印迹后,在所有胎盘匀浆和培养基中均检测到β-HCG,但在任何相应的卵黄囊组织样本中均未检测到。逆转录聚合酶链反应(RT-PCR)显示,所有胎盘样本均表达β-HCG mRNA,而所有卵黄囊和肝脏样本均表达αFP mRNA。这些发现表明,卵黄囊膜是胚外和胚胎区室之间重要的转运区域,也可能有助于进一步开发利用胎儿体细胞基因治疗的治疗方案,即将转导细胞注入胚外体腔。