Distribution of transforming growth factor alpha precursors in the mouse uterus during the periimplantation period and after steroid hormone treatments.
Distribution of transforming growth factor alpha precursors in the mouse uterus during the periimplantation period and after steroid hormone treatments.
复制标题
在植入期间和类固醇激素治疗后，转化生长因子α前体在小鼠子宫中的分布。
DOI：
10.1095/biolreprod50.3.481
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发表时间：
1994
影响因子：
3.6
通讯作者：
Dey,SK
中科院分区：
文献类型：
--
作者：
Paria,BC;Das,SK;Huet-Hudson,YM;Dey,SK
采用免疫组织化学方法，使用识别生长因子前体形式的抗体，检测了围着床期小鼠子宫中转化生长因子α（TGF α）前体（proTGF α）的时间和细胞类型特异性分布，以及卵巢切除成年小鼠类固醇激素治疗后的子宫中转化生长因子α（TGF α）前体（proTGF α）的时间和细胞类型特异性分布。这些研究通过分离的子宫细胞型制剂中proTGF α的免疫印迹分析进行补充。通过使用转基因小鼠的胰腺或泌乳乳腺证实了这些研究中使用的抗体的特异性，其中突变的proTGF α（缺乏蛋白水解裂解的识别位点）在组织特异性增强子/启动子下靶向表达。组织化学研究分析显示，在妊娠或假性妊娠第1天，免疫反应性proTGF α主要在管腔和腺上皮细胞中蓄积，随后在第2天和第3天几乎没有蓄积。然而，免疫反应性proTGF α在第4天上午开始在腔上皮中重新出现，并在下午变得更加突出。在妊娠小鼠中，在附着反应期间（第4天2130 h），植入部位的这些细胞中的免疫染色持续存在，但在第5天早晨消失。免疫染色似乎位于腔上皮的顶端边缘。在假孕第5或6天的非受体子宫中未检测到阳性免疫染色。与免疫组化结果一致，Western印迹分析检测到两种前体蛋白（14.5和17 kDa）在分离的管腔上皮细胞富集制剂的第4天，但不是在第5天，假孕。结果表明，proTGF α在着床前在受体子宫的腔上皮中积累。用免疫组化和免疫印迹法检测卵巢类固醇对卵巢切除成年小鼠子宫内proTGF α积聚的影响。而注射雌二醇-17 β（E2）或孕酮（P4）对proTGF α的上皮积累几乎没有或适度的影响，P4引发数天导致proTGF α在上皮细胞中的明显积累。叠加的E2治疗P4引发表现出双相反应，通过12小时的免疫染色的初始逐渐损失，然后由24小时的E2治疗的回报。本研究所采用的联合激素治疗方案类似于P4致敏的延迟着床小鼠用E2诱导着床的情况。结果表明，旁分泌/“altacrine”的作用，这种生长因子在植入。
Temporal and cell-type specific distribution of transforming growth factor α (TGF α) precursor (proTGF α) was examined in the mouse uterus during the periimplantation period, and after steroid hormone treatments of ovariectomized adult mice by immunohistochemistry using antibodies that recognize the precursor forms of the growth factor. These studies were complemented by immunoblot analysis of proTGF α in separated uterine cell-type preparations. The specificity of the antibodies used in these studies was confirmed by use of pancreas or lactating mammary glands from transgenic mice in which mutated proTGF α, lacking recognition sites for proteolytic cleavages, was targeted for expression under a tissue-specific enhancer/promoter. Analysis of histochemical studies revealed accumulation of immunoreactive proTGF α primarily in luminal and glandular epithelial cells on Day 1 of pregnancy or pseudopregnancy followed by little or no accumulation on Days 2 and 3. However, immunoreactive proTGF α started to reappear in the luminal epithelium on the morning of Day 4 and became more prominent in the afternoon. In pregnant mice, immunostaining persisted in these cells at the implantation sites during the time of attachment reaction (2130 h on Day 4), but disappeared by morning of Day 5. Immunostaining appeared to be situated at the apical border of the luminal epithelium. No positive immunostaining could be detected in the nonreceptive uterus on Day 5 or 6 of pseudopregnancy. Consistent with the immunohistochemistry results, Western blot analysis detected two species of precursor proteins (14.5 and 17 kDa) in isolated luminal epithelial cell-enriched preparations on Day 4, but not on Day 5, of pseudopregnancy. The results suggest that proTGF α accumulates in the luminal epithelium of the receptive uterus prior to implantation. The effects of ovarian steroids on uterine accumulation of proTGF α were examined in ovariectomized adult mice by immunohistochemistry and immunoblotting. Whereas an injection of estradiol-17 β (E2) or progesterone (P4) had little or a modest effect on epithelial accumulation of proTGF α, P4priming for several days resulted in distinct accumulation of proTGF α in epithelial cells. The superimposition of an E2treatment on P4priming showed a biphasic response, with an initial gradual loss of immunostaining through 12 h followed by a return by 24 h of E2treatment. The combined hormone treatment schedule employed here is similar to the situation of inducing implantation with E2in P4-primed delayed implanting mice. The results suggest a paracrine/“juxtacrine” role for this growth factor in implantation.