Alsat E, Bouali Y, Goldstein S, Malassiné A, Berthelier M, Mondon F, Cedard L
Mol Cell Endocrinol. 1984 Dec;38(2-3):197-203. doi: 10.1016/0303-7207(84)90118-7.
Microvillous membranes isolated from early gestation placentas (8-12 weeks of amenorrhoea) and from mid-term placentas (20-22 weeks of amenorrhoea) were used to study the specific binding of low-density lipoprotein (LDL) to the trophoblast. The purity of the microvillous preparations has been assessed by electron microscopy and by their enrichment in two membrane markers, 5'-nucleotidase and alkaline phosphatase. Evidence was presented demonstrating the existence of saturable binding sites for [125I]LDL in placental microvilli as early as 6 weeks of pregnancy. The apparent KD values for these binding sites have been determined by Scatchard analyses to be 6.98 +/- 0.83 and 6.57 +/- 0.81 micrograms protein LDL/ml, for early gestation and mid-term preparations, respectively. This apparent KD value was unaffected by a pretreatment of the membranes by heparin, as indicated by the mean values of 7.13 +/- 0.89 and 6.97 +/- 0.75 micrograms protein LDL/ml obtained for immature microvilli preincubated with or without heparin, respectively. Large variations of binding capacity were observed in each gestational age group and no significant difference was found between them. These results indicate that the LDL binding sites of the human placenta, located on the microvillous membranes, (i) are present as early as the 6th week of pregnancy, and (ii) display the same high affinity and specificity for LDL as those of the term trophoblast.
从妊娠早期胎盘(闭经8 - 12周)和中期胎盘(闭经20 - 22周)分离出的微绒毛膜,用于研究低密度脂蛋白(LDL)与滋养层细胞的特异性结合。通过电子显微镜以及两种膜标志物5'-核苷酸酶和碱性磷酸酶的富集情况,评估了微绒毛制剂的纯度。有证据表明,早在妊娠6周时,胎盘微绒毛中就存在[125I]LDL的饱和结合位点。通过Scatchard分析确定,这些结合位点的表观解离常数(KD)值,对于妊娠早期制剂和中期制剂,分别为6.98±0.83和6.57±0.81微克蛋白质LDL/毫升。如分别对未成熟微绒毛进行肝素预处理(有或无)后得到的平均值7.13±0.89和6.97±0.75微克蛋白质LDL/毫升所示,该表观KD值不受肝素对膜预处理的影响。在每个孕周组中均观察到结合能力的较大差异,且各孕周组之间未发现显著差异。这些结果表明,位于微绒毛膜上的人胎盘LDL结合位点:(i)早在妊娠第6周就已存在;(ii)对LDL表现出与足月滋养层细胞相同的高亲和力和特异性。