STAGE-SPECIFIC ALTERATION IN THE APICAL MEMBRANE-GLYCOPROTEINS OF ENDROMETRIAL EPITHELIAL-CELLS RELATED TO IMPLANTATION IN RABBITS
STAGE-SPECIFIC ALTERATION IN THE APICAL MEMBRANE-GLYCOPROTEINS OF ENDROMETRIAL EPITHELIAL-CELLS RELATED TO IMPLANTATION IN RABBITS
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DOI：
10.1095/biolreprod34.4.701
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发表时间：
1986-05-01
影响因子：
3.6
通讯作者：
HOFFMAN, LH
中科院分区：
文献类型：
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作者：
ANDERSON, TL;OLSON, GE;HOFFMAN, LH
兔子子宫内膜上皮在囊胚着床之前经历分化，包括表面负性的丧失和糖萼形态的变化。使用非怀孕（动情）和假怀孕的兔子来研究管腔上皮膜及其糖萼的蛋白质和糖组成的特定变化，这些变化与植入接受性的获得有关。怀孕的动物被用来研究通过植入囊胚对管腔表面的进一步修饰。通过在含有蛋白酶抑制剂的 1% Triton X-100 的管腔内孵育 15 分钟来溶解管腔上皮细胞的顶端表面。通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）分离提取液中的蛋白质。在接受兔的子宫内鉴定出三种新的多肽（24 kDa、42 kDa 和 58 kDa）。琥珀酰麦芽凝集素 (sWGA) 和蓖麻凝集素 (RCA-I) 凝集素与通过 SDS-PAGE 分离的提取物的蛋白质印迹上的 24 kDa 和 42 kDa 成分结合，将其鉴定为糖蛋白。此外，其他多肽（26 kDa、80-86 kDa 和 145 kDa）显示出对 WGA、RCA-I 或刀豆球蛋白 A (Con A) 的亲和力变化，具体取决于激素状态。与这些发现相关的是，与发情动物相比，这些凝集素与受体兔子子宫中完整非纤毛细胞的结合增加；无论激素状况如何，纤毛细胞都会特异性结合双花扁豆凝集素 (DBA)。在凝集素暴露之前用神经氨酸酶处理发情动物的子宫，或对这些动物的蛋白质进行蛋白质印迹，表明在非接受性兔子中存在具有唾液酸-D-半乳糖末端的糖蛋白。与非植入位点相比，注意到凝集素与植入位点的完整细胞以及从这些位点分离的蛋白质印迹的结合减少。这些结果为植入前子宫内膜上皮顶端表面的蛋白质和糖组成的阶段特异性改变提供了证据，并表明植入囊胚进一步改变了管腔表面。
The rabbits endometrial epithelium undergoes differentiation prior to the time of blastocyst implatnation, including loss of surface negativity and a change in glycocalyx morphology. Nonpregnant (estrous) and pseudo-pregnant rabbits were used to study specific alterations in proteins and saccharide composition of the luminal epithelial membrane and its glycocalyx related to the acquisitions of receptivity to implantation. Pregnant animals were used to study further modification of the luminal surface by implanting blastocysts. The apical surface of luminal epithelial cells was solubilized by a 15-min intraluminal incubation of 1% Triton X-100 containing protease inhibitors. Proteins in extract solutions were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Three new polypeptides (24 kDa, 42 kDa and 58 kDa) were identified in uteri from receptive rabbits. Binding of succinyl Wheat Germ Agglutinin (sWGA) and Ricinus communis Agglutinin (RCA-I) lectins to the 24 kDa and 42 kDa components on Western blots of extracts separated by SDS-PAGE identified them as glycoproteins. Additionally, other polypeptides (26 kDa, 80-86 kDa and 145 kDa) showed changes in affinity for WGA, RCA-I or concanavalin A (Con A), depending on the hormonal state. Correlating with these findings was an increased binding of these lectins to intact nonciliated cells in uteri of receptive rabbits compared to estrous animals; ciliated cells bound Dolichos biflorus Agglutinin (DBA) specifically, regardless of the hormonal condition. Treatment of uteri from estrous animals, or Western blots of proteins from these animals, with neuraminidase prior to lectin exposure suggested the presence of glycoproteins having a sialic acid-D-galactose terminus in nonreceptive rabbits. Reduced binding of lectin to intact cells at implantation sites and to blots of proteins isolated from these sites, compared to nonimplantation sites, was noted. These results provide evidence for stage-specific alterations in protein and saccharide composition of the apical surface of endometrial epithelium prior to implantation, and indicate that implanting blastocysts further modify the luminal surface.