Cell interactions with laminin and its proteolytic fragments during outgrowth of mouse primary trophoblast cells.
Cell interactions with laminin and its proteolytic fragments during outgrowth of mouse primary trophoblast cells.
复制标题
小鼠原代滋养层细胞生长过程中细胞与层粘连蛋白及其蛋白水解片段的相互作用。
DOI：
10.1095/biolreprod45.5.664
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发表时间：
1991
影响因子：
3.6
通讯作者：
Armant,DR
中科院分区：
文献类型：
--
作者：
Armant,DR
小鼠囊胚在无血清培养24 - 48小时后，变得具有附着能力，附着在纤连蛋白或层粘连蛋白包被的表面上，随后形成滋养层生长。胚泡层粘连蛋白受体的特点是在生长研究中使用修饰的层粘连蛋白。滋养层细胞与层粘连蛋白的肽部分相互作用，但不与寡糖部分相互作用，因为其粘附活性通过煮沸或胰蛋白酶处理而降低，但不通过去除或修饰其碳水化合物的处理。通过使用层粘连蛋白E1−4和E8的充分表征的蛋白水解片段以及合成肽CDPGYIGSR，层粘连蛋白生长促进活性进一步定位于其结构域。层粘连蛋白的E1 - 4片段不能促进胚胎生长。然而，E8片段，其中含有肝素结合域以及在细胞粘附和神经突生长过程中识别的网站，大力促进生长在肝素，硫酸乙酰肝素，或肝素酶的存在和不存在。与这些结果一致，完整层粘连蛋白上的生长不受CDPGYIGSR的抑制，CDPGYIGSR是已知介导某些细胞类型粘附的E1−4片段内的序列。从这些结果可以得出结论，早期滋养层细胞粘附层粘连蛋白的E8结构域中的肽使用的机制是独立的用于粘附到纤连蛋白。
Mouse blastocysts in serum-free culture for 24−48 h become attachment-competent, adhere to fibronectin- or laminin-coated surfaces, and subsequently form trophoblast outgrowths. The blastocyst laminin receptor was characterized in outgrowth studies using modified laminin. Trophoblast cells interacted with the peptide portion of laminin, but not the oligosaccharide moiety since its adhesive activity was reduced by boiling or trypsin treatment, but not by treatments that removed or modified its carbohydrate. Laminin outgrowth-promoting activity was further localized within its structural domains by use of the well-characterized proteolytic fragments of laminin, E1−4, and E8, and a synthetic peptide, CDPGYIGSR. The E1−4 fragment of laminin did not promote embryo outgrowth. However, the E8 fragment, which contains a heparin-binding domain as well as sites recognized during cell adhesion and neurite outgrowth, vigorously promoted outgrowth in both the presence and absence of heparin, heparan sulfate, or heparinase. Consistent with these results, outgrowth on intact laminin was not inhibited by CDPGYIGSR, a sequence within the E1−4 fragment that is known to mediate the adhesion of some cell types. It is concluded from these results that early trophoblast cells adhere to peptide in the E8 domain of laminin using a mechanism that is independent of the one used for adhesion to fibronectin.