X-linked lymphocyte regulated gene 5c-like (Xlr5c-like) is a novel target of progesterone action in granulosa cells of periovulatory rat ovaries.
X-linked lymphocyte regulated gene 5c-like (Xlr5c-like) is a novel target of progesterone action in granulosa cells of periovulatory rat ovaries.
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DOI：
10.1016/j.mce.2015.05.008
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发表时间：
2015-09-05
影响因子：
4.1
通讯作者：
Jo M
中科院分区：
文献类型：
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作者：
Mishra B;Park JY;Wilson K;Jo M
黄体酮（P4）通过其核受体（PGR）发挥作用，通过介导与排卵和/或黄体形成有关的基因表达，在排卵中发挥重要作用。为了鉴定排卵特异性PGR调控基因，用hCG±RU486 （PGR拮抗剂）处理的大鼠颗粒细胞进行了初步的微阵列分析。RU486下调最多的转录本是EST（表达序列标签）序列（gb: BI289578.1），该序列与xlr505样mRNA的预测序列相匹配。由于对Xlr5c-like一无所知，我们首先表征了Xlr5c-like mRNA在大鼠卵巢中的表达模式。pmsg刺激大鼠卵巢促性腺激素（hCG）后，大鼠卵泡颗粒细胞中Xlr5c-like mRNA水平瞬间上调。在体外培养的排卵前卵巢颗粒细胞中，hCG处理可模拟xlr5c样mRNA的瞬时诱导。我们进一步证明，lh激活的PKA、MEK、PI3K和p38信号通路参与了xlr5c样mRNA的增加。xlr5c样mRNA的增加被RU486所消除。MPA（合成黄体酮）可逆转RU486的抑制作用，地塞米松（合成糖皮质激素）则不能。此外，Xlr5c-like启动子区SP1/SP3和PGR应答元件位点的突变降低了Xlr5c-like报告基因的活性。RU486也抑制xlr5c样报告基因的活性。ChIP实验证实了PGR和SP3在排卵期颗粒细胞中与xlr5c样启动子结合。在功能上，在颗粒细胞培养中sirna介导的xlr5c样敲低导致Snap25、Cxcr4和Adamts1的mRNA水平降低。利用腺病毒方法表达的重组xlr5c样蛋白主要定位于细胞核，在较小程度上定位于大鼠颗粒细胞的细胞质。综上所述，本研究首次报道了hCG在大鼠排卵期卵巢中调控xlr5c样mRNA的时空表达。P4/PGR介导了lh诱导的xlr5c样mRNA的增加。反过来，Xlr5c-like参与调节特定排卵基因的表达，如Snap25、Cxcr4和Adamts1，可能在排卵周颗粒细胞的细胞核中起作用。
Progesterone (P4), acting through its nuclear receptor (PGR), plays an essential role in ovulation by mediating the expression of genes involved in ovulation and/or luteal formation. To identify ovulatory specific PGR-regulated genes, a preliminary microarray analysis was performed using rat granulosa cells treated with hCG ± RU486 (PGR antagonist). The transcript most highly down-regulated by RU486 was an EST (Expressed Sequence Tag) sequence (gb: BI289578.1) that matches with predicted sequence for Xlr5c-like mRNA. Since nothing is known about Xlr5c-like, we first characterized the expression pattern of Xlr5c-like mRNA in the rat ovary. The level of mRNA for Xlr5c-like is transiently up-regulated in granulosa cells of periovulatory follicles after hCG stimulation in PMSG-primed rat ovaries. The transient induction of Xlr5c-like mRNA was mimicked by hCG treatment in cultured granulosa cells from preovulatory ovaries. We further demonstrated that the LH-activated PKA, MEK, PI3K, and p38 signaling is involved in the increase in Xlr5c-like mRNA. The increase in Xlr5c-like mRNA was abolished by RU486. The inhibitory effect of RU486 was reversed by MPA (synthetic progestin), but not by dexamethasone (synthetic glucocorticoid). Furthermore, mutation of SP1/SP3 and PGR response element sites in the promoter region of Xlr5c-like decreased Xlr5c-like reporter activity. RU486 also inhibited Xlr5c-like reporter activity. ChIP assay verified the binding of PGR and SP3 to the Xlr5c-like promoter in periovulatory granulosa cells. Functionally, siRNA-mediated Xlr5c-like knockdown in granulosa cell cultures resulted in reduced levels of mRNA for Snap25, Cxcr4, and Adamts1. Recombinant Xlr5c-like protein expressed using an adenoviral approach was localized predominantly to the nucleus and to a lesser extent to the cytoplasm of rat granulosa cells. In conclusion, this is the first report showing the spatiotemporally regulated expression of Xlr5c-like mRNA by hCG in rat periovulatory ovaries. P4/PGR mediates the LH-induced increase in Xlr5c-like mRNA. In turn, Xlr5c-like is involved in regulating the expression of specific ovulatory genes such as Snap25, Cxcr4, and Adamts1, possibly acting in the nucleus of periovulatory granulosa cells.
DOI：
10.1084/jem.166.6.1702
发表时间：
1987-12-01
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Siegel JN;Turner CA;Klinman DM;Wilkinson M;Steinberg AD;MacLeod CL;Paul WE;Davis MM;Cohen DI
通讯作者：
Cohen DI