Localization of estrogen receptor messenger ribonucleic acid in rhesus monkey uterus by nonradioactive in situ hybridization with digoxigenin-labeled oligodeoxynucleotides.
Localization of estrogen receptor messenger ribonucleic acid in rhesus monkey uterus by nonradioactive in situ hybridization with digoxigenin-labeled oligodeoxynucleotides.
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DOI：
10.1210/endo.132.1.8419136
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发表时间：
1993
期刊：
影响因子：
4.8
通讯作者：
Takehiko Koji;R. Brenner
中科院分区：
文献类型：
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作者：
Takehiko Koji;R. Brenner
以前的免疫细胞化学研究表明，受体调节不同类型的子宫细胞。在灵长类动物中，孕酮（P）抑制功能区腺上皮细胞中的雌激素受体（ER），但不能抑制基底区腺上皮（GE）细胞中的ER。P也不能抑制ER在血管周围的基质和平滑肌细胞的螺旋动脉在functionalis。我们使用非放射性原位杂交来确定是否类似的细胞类型的差异发生在ER mRNA水平。我们使用地高辛标记的寡脱氧核苷酸（oligo-DNAs; 45-mer）作为探针，并检测杂交瘤与辣根过氧化物酶标记的抗地高辛抗体的免疫细胞化学。该技术可以区分紧密组织学组织中的阳性和阴性细胞。在去势猴中，大多数GE细胞以及子宫内膜间质细胞ER mRNA阳性，而所有血管平滑肌、内皮细胞和血管周围间质细胞均为阴性。雌激素处理14天后，GE细胞、大部分间质细胞、螺旋动脉血管平滑肌和血管周围间质细胞的ER mRNA表达明显增加。然而，在基底部，螺旋动脉的这些成分是阴性的，基底部的小基底动脉也是阴性的。在大多数阳性细胞中，ER mRNA不是均匀分布在细胞质中，而是集中在其核周区域。GE细胞在基底有特别强烈的浓度在其顶端的核周信号。经雌二醇和P处理后，功能肌的GE细胞信号明显减弱，而基底肌的GE细胞、螺旋动脉的血管平滑肌细胞和血管周围基质细胞信号不减弱。在子宫肌层中，ER mRNA定位于平滑肌细胞的核周区域，但染色强度不受激素操作的显著影响。出乎意料的是，我们在子宫内膜/子宫肌层边缘和子宫肌层结缔组织更深处观察到以极高的ER mRNA阳性信号为特征的基质细胞簇（“热细胞”），尽管这些细胞不表达高水平的ER蛋白。然而，一般而言，ER mRNA的细胞分布及其激素调节不同于ER蛋白。
Previous immunocytochemical studies indicate that receptor regulation varies in different uterine cell types. In primates, progesterone (P) suppresses estrogen receptor (ER) in glandular epithelial cells in the functionalis, but fails to suppress ER in the glandular epithelial (GE) cells of the basalis. P also fails to suppress ER in the perivascular stromal and smooth muscle cells of the spiral arteries in the functionalis. We used nonradioactive in situ hybridization to determine whether similar cell type differences occur at the ER mRNA level. We used digoxigenin-labeled oligodeoxynucleotides (oligo-DNAs; 45-mer) as probes and detected the hybrids immunocytochemically with horseradish peroxidase-labeled antidigoxigenin antibody. This technique can discriminate between positive and negative cells in closely packed histological associations. In spayed monkeys, most of the GE cells as well as endometrial stromal cells were positive for ER mRNA, while all vascular smooth muscle, endothelium, and perivascular stromal cells were negative. Estradiol treatment for 14 days markedly increased ER mRNA staining in the GE cells, most stromal cells, and the vascular smooth muscle and perivascular stromal cells of spiral arteries in the functionalis. However, in the basalis, these components of the spiral arteries were negative as were the small basal arteries of the basalis. In most positive cells, ER mRNA was not homogeneously distributed in the cytoplasm, but, rather, was concentrated in their perinuclear regions. The GE cells in the basalis had especially intense concentrations of perinuclear signal at their apical poles. After sequential estradiol plus P treatment, the signal was greatly reduced in the GE cells of the functionalis, but not in the GE cells of the basalis or in the vascular smooth muscle or perivascular stromal cells of the spiral arteries of the functionalis. In myometrium, ER mRNA was localized to the perinuclear region of smooth muscle cells, but the staining intensity was not dramatically affected by hormonal manipulation. Unexpectedly, we observed clusters of stromal cells characterized by extremely high positive signals for ER mRNA ("hot cells") at the endometrial/myometrial border and deeper in the connective tissue of the myometrium, although such cells did not express high levels of ER protein. In general, however, the cellular distribution of ER mRNA and its hormonal regulation paralleled those of ER protein.