Farrar J D, Carson D D
University of Texas M.D. Anderson Cancer Center, Department of Biochemistry and Molecular Biology, Houston 77030.
Biol Reprod. 1992 Jun;46(6):1095-108. doi: 10.1095/biolreprod46.6.1095.
Changes in the temporal and spatial patterns of expression of mRNA encoding uterine extracellular matrix (ECM) proteins were determined during the peri-implantation period. Northern blot hybridization of cDNAs corresponding to laminin (LM) B1, LM B2, entactin, fibronectin, collagen (CL) type IV alpha 1, and CL IV alpha 2 was performed on RNA extracted from either whole mouse uteri or endometrial explants between Day 4, i.e., the day of implantation, and Day 7 of pregnancy, when the decidual response is well established. These analyses revealed a dramatic increase in LM B2, CL IV alpha 1, and CL IV alpha 2 mRNA expression by Day 7 of pregnancy. Relative levels of the mRNA encoding other ECM components, including LM B1, were not altered when compared to changes in the relative level of expression of glyceraldehyde-3-phosphate dehydrogenase mRNA. The differential expression of the B chains of LM appeared to be limited to the stromal cells of the endometrium. In situ hybridization of uterine sections with cRNA probes corresponding to LM B1, LM B2, and CL IV alpha 1 demonstrated that LM B1 was expressed temporally in high amounts in the primary decidual zones (PDZ) and persisted throughout PDZ degeneration. LM B2 mRNA was expressed in both primary and secondary decidual zones and persisted through Day 8 of pregnancy. CL IV alpha 1 mRNA expression mimicked that of LM B2. Oviduct ligation on Day 2 of pregnancy was used to prevent embryo transport to one uterine horn, whereas decidualization and embryo implantation were permitted in the contralateral horn. This experiment demonstrated that the increases in uterine ECM mRNA expression were not due solely to the changing hormonal milieu of the uterus. ECM components, including CL IV, have been shown to bind growth factors such as transforming growth factor-beta (TGF-beta) in an insoluble but biologically active form. The remarkable similarity between the pattern of CL IV and LM B2 expression and previously reported TGF-beta deposition (Tamada et al., Mol Endocrinol 1990; 4:965-972) prompted examination of the effects of this growth factor on blastocyst development in vitro. TGF-beta 1 was tested for its ability to alter embryo outgrowth on LM-coated tissue culture surfaces; however, significant differences in the rate or extent of outgrowth in the presence of TGF-beta were not detected.(ABSTRACT TRUNCATED AT 250 WORDS)
在围植入期,确定了编码子宫细胞外基质(ECM)蛋白的mRNA表达的时空模式变化。在妊娠第4天(即植入日)至妊娠第7天(此时蜕膜反应已充分建立)之间,对从整个小鼠子宫或子宫内膜外植体中提取的RNA进行了与层粘连蛋白(LM)B1、LM B2、巢蛋白、纤连蛋白、IV型胶原蛋白(CL)α1和CL IV α2相对应的cDNA的Northern印迹杂交。这些分析显示,到妊娠第7天,LM B2、CL IV α1和CL IV α2 mRNA表达显著增加。与甘油醛-3-磷酸脱氢酶mRNA表达的相对水平变化相比,编码其他ECM成分(包括LM B1)的mRNA的相对水平没有改变。LM B链的差异表达似乎仅限于子宫内膜的基质细胞。用与LM B1、LM B2和CL IV α1相对应的cRNA探针进行子宫切片的原位杂交表明,LM B1在初级蜕膜区(PDZ)中大量短暂表达,并在整个PDZ退化过程中持续存在。LM B2 mRNA在初级和次级蜕膜区均有表达,并持续到妊娠第8天。CL IV α1 mRNA表达与LM B2相似。在妊娠第2天进行输卵管结扎,以防止胚胎运输到一个子宫角,而对侧子宫角允许蜕膜化和胚胎植入。该实验表明,子宫ECM mRNA表达的增加并非仅由于子宫激素环境的变化。已证明包括CL IV在内的ECM成分能以不溶性但具有生物活性的形式结合生长因子,如转化生长因子-β(TGF-β)。CL IV和LM B2表达模式与先前报道的TGF-β沉积模式(Tamada等人,《分子内分泌学》1990年;4:965 - 972)之间的显著相似性促使人们研究这种生长因子对体外胚泡发育 的影响。测试了TGF-β1改变在LM包被的组织培养表面上胚胎生长的能力;然而,未检测到在有TGF-β存在时生长速率或程度的显著差异。(摘要截短于250字)