Localization of ISG15 and conjugated proteins in bovine endometrium using immunohistochemistry and electron microscopy
Localization of ISG15 and conjugated proteins in bovine endometrium using immunohistochemistry and electron microscopy
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DOI：
10.1210/en.2003-1087
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发表时间：
2004-02-01
期刊：
影响因子：
4.8
通讯作者：
Hansen, TR
中科院分区：
文献类型：
--
作者：
Austin, KJ;Carr, AL;Hansen, TR
干扰素刺激的基因 ISG15（一种泛素同源物）在妊娠第 18 天与孕体来源的干扰素 tau 蛋白结合并调节子宫蛋白。这里假设 ISG15 在子宫内膜细胞内的细胞定位可能提供有关功能的见解。在发情周期和怀孕的第17-21/0天以及怀孕的第23、45和50天从奶牛（类似于21天的动情周期）收集子宫。细胞内 ISG15 及其结合物在妊娠第 17 天出现，在第 18 至 23 天达到最高水平，然后根据蛋白质印迹在第 45 天降至较低但可检测的水平（P < 0.05）。在妊娠第 50 天或动情周期期间未检测到 ISG15 及其结合物。免疫组织化学显示，ISG15 在第 18-23 天定位于整个子宫内膜，其中腔下致密层和整个海绵层的腺上皮染色最重。到第 45 和 50 天时，ISG15 仅在紧邻管腔上皮下方的致密层中被轻微染色。使用透射电子显微镜和免疫金标记，ISG15 特异性定位于子宫内膜上皮细胞和基质细胞的细胞器和区室：细胞核、核周间隙、细胞质、线粒体、粗面内质网和细胞膜。这种在上皮细胞和基质细胞中的特异性定位得出这样的结论：ISG15 具有多种细胞内功能。妊娠第 50 天期间缀合 ISG15 的持续存在可能反映了缀合蛋白对植入和胎盘发育的反应的稳定性。
The interferon-stimulated gene ISG15, a ubiquitin homolog, becomes conjugated to and regulates uterine proteins in response to conceptus-derived interferon-tau on d 18 of pregnancy. It was hypothesized here that cellular localization of ISG15 within endometrial cells might provide insight regarding function. Uteri were collected from cows (similar to21-d estrous cycle) on d 17-21/0 of the estrous cycle and pregnancy and d 23, 45, and 50 of pregnancy. Intracellular ISG15 and its conjugates were present on d 17 of pregnancy, peaked to highest levels from d 18 to 23 and then declined to low but detectable levels by d 45 (P < 0.05) based on Western blotting. ISG15 and its conjugates were not detected on d 50 of pregnancy or during the estrous cycle. Immunohistochemistry revealed that ISG15 was localized throughout the endometrium on d 18-23, with heaviest staining in the sublumenal stratum compactum and the glandular epithelium throughout the stratum spongiosum. By d 45 and 50, ISG15 was lightly stained only in the stratum compactum immediately beneath the lumenal epithelium. Using transmission electron microscopy and immunogold labeling, ISG15 was specifically localized to organelles and compartments of endometrial epithelial cells and stromal cells: nucleus, perinuclear space, cytosol, mitochondria, rough endoplasmic reticulum, and cell membrane. This specific localization in epithelial and stromal cells led to the conclusion that ISG15 has diverse intracellular functions. The sustained presence of conjugated ISG15 through d 50 of pregnancy might reflect stabilization of conjugated proteins in response to implantation and the development of the placenta.