Localization of oxytocin receptor mRNA in the ovine uterus during the oestrous cycle and early pregnancy.
Localization of oxytocin receptor mRNA in the ovine uterus during the oestrous cycle and early pregnancy.
复制标题
发情周期和妊娠早期催产素受体 mRNA 在绵羊子宫中的定位。
DOI：
--
复制
发表时间：
1994
影响因子：
3.5
通讯作者：
D. Wathes
中科院分区：
文献类型：
--
作者：
K. Stevenson;P R Riley;H. Stewart;A. P. Flint;D. Wathes
将合成的与部分绵羊子宫内膜催产素受体cDNA相对应的45-mer寡核苷酸杂交到40只母羊的子宫切片上，这些母羊分别处于发情周期、妊娠前3周和季节性无情期的不同阶段。采用图像分析方法，以光密度（OD）值测量放射线自影片上催产素受体mRNA的数量。在周期的第14-15天，信息首先出现在腔上皮中，在发情期增加到0.48的峰值OD，在第2 - 5天再次下降。在发情前第15天，浅腺体、深腺体和环状间质中的催产素受体mRNA升高，OD值峰值分别为0.17、0.11和0.11，第2天再次下降，第5天达到基础值（OD < 0.015）。对肌层的杂交从第2 ~ 15天的平均OD值0.01上升到第0 ~ 1天的峰值0.03 +/- 0.01（平均+/- S.E.M.），但变化不显著。在怀孕母羊中，在第14-15天，在任何区域都没有检测到催产素受体mRNA，但在第21天，3只母羊中的2只出现了腔上皮杂交。在不发情的母羊中，子宫内膜所有区域的催产素受体mRNA浓度约为发情时的一半。将不同子宫间室中催产素受体mRNA的光密度读数与使用125i标记的催产素拮抗剂d(CH2)5[Tyr(Me)2,Thr4,Tyr- nh2(9)]-血管催产素（125i标记的OTA）结合研究评估的相同区域的催产素受体测量值进行比较。在子宫内膜中，受体mRNA和125i标记的OTA结合模式平行变化，两组测量值均显著相关（P < 0.01）。在肌层，125i标记的OTA结合在发情期显著增加；这并没有伴随着催产素受体mRNA杂交的类似增加。这项研究有助于证实先前鉴定的cDNA克隆来源于绵羊催产素受体，因为子宫内膜中催产素受体mRNA的表达模式与催产素结合的模式非常相似。最大的表达和结合都发生在发情期，这表明子宫中催产素受体基因的调节主要发生在转录水平，而不是在翻译水平。未检测到发情期子宫内膜mRNA表达的显著增加可能表明子宫内膜和子宫内膜催产素受体具有不同的亚型。
A synthetic 45-mer oligonucleotide corresponding to part of the ovine endometrial oxytocin receptor cDNA was hybridized to sections of ovine uterus collected from 40 ewes at different stages during the oestrous cycle, the first 3 weeks of pregnancy and seasonal anoestrus. The quantity of oxytocin receptor mRNA was measured as the optical density (OD) value on autoradiographs using image analysis. Message first appeared in the luminal epithelium on days 14-15 of the cycle, increasing to a peak OD of 0.48 at oestrus and decreasing again between days 2 and 5. Oxytocin receptor mRNA in the superficial glands, deep glands and caruncular stroma increased between day 15 and oestrous to peak OD values of 0.17, 0.11 and 0.11 respectively, declining again by day 2 and reaching basal values (OD < 0.015) by day 5. Hybridization to the myometrium tended to rise from a mean OD value of 0.01 on days 2-15 to a peak of 0.03 +/- 0.01 (mean +/- S.E.M.) on days 0-1, but the change was not significant. In pregnant ewes there was no detectable oxytocin receptor mRNA on days 14-15 in any region, but hybridization to the luminal epithelium was present in two of three ewes on day 21. In anoestrous ewes oxytocin receptor mRNA concentrations in all areas of the endometrium were approximately half those measured at oestrus. Optical density readings for oxytocin receptor mRNA in the various uterine compartments were compared with measurements of oxytocin receptors in the same regions as assessed by binding studies using the 125I-labelled oxytocin antagonist d(CH2)5[Tyr(Me)2,Thr4,Tyr-NH2(9)]-vasotocin (125I-labelled OTA). In the endometrium, receptor mRNA and 125I-labelled OTA binding patterns changed in parallel, and both sets of measurements were significantly correlated (P < 0.01). In the myometrium, a significant increase in 125I-labelled OTA binding occurred at oestrus; this was not accompanied by a similar increase in oxytocin receptor mRNA hybridization. This study helps to confirm that the previously identified cDNA clone is derived from the ovine oxytocin receptor, as patterns of oxytocin receptor mRNA expression in the endometrium closely resembled those of oxytocin binding. Maximum expression and binding both occurred at oestrus, suggesting that regulation of the oxytocin receptor gene in the uterus occurs principally at the transcriptional, rather than at the translational, level. Failure to detect a significant increase in myometrial mRNA expression at oestrus may indicate that the endometrial and myometrial oxytocin receptors are of different isoforms.